Neuroserpin,a secreted protein that belongs to the serpin superfamily of serine protease inhibitors,is highly expressed in the central nervous system and plays multiple roles in brain development and pathology.As a na...Neuroserpin,a secreted protein that belongs to the serpin superfamily of serine protease inhibitors,is highly expressed in the central nervous system and plays multiple roles in brain development and pathology.As a natural inhibitor of recombinant tissue plasminogen activator,neuroserpin inhibits the increased activity of tissue plasminogen activator in ischemic conditions and extends the therapeutic windows of tissue plasminogen activator for brain ischemia.However,the neuroprotective mechanism of neuroserpin against ischemic stroke remains unclear.In this study,we used a mouse model of middle cerebral artery occlusion and oxygen-glucose deprivation/reperfusion-injured cortical neurons as in vivo and in vitro ischemia-reperfusion models,respectively.The models were used to investigate the neuroprotective effects of neuroserpin.Our findings revealed that endoplasmic reticulum stress was promptly triggered following ischemia,initially manifesting as the acute activation of endoplasmic reticulum stress transmembrane sensors and the suppression of protein synthesis,which was followed by a later apoptotic response.Notably,ischemic stroke markedly downregulated the expression of neuroserpin in cortical neurons.Exogenous neuroserpin reversed the activation of multiple endoplasmic reticulum stress signaling molecules,the reduction in protein synthesis,and the upregulation of apoptotic transcription factors.This led to a reduction in neuronal death induced by oxygen/glucose deprivation and reperfusion,as well as decreased cerebral infarction and neurological dysfunction in mice with middle cerebral artery occlusion.However,the neuroprotective effects of neuroserpin were markedly inhibited by endoplasmic reticulum stress activators thapsigargin and tunicamycin.Our findings demonstrate that neuroserpin exerts neuroprotective effects on ischemic stroke by suppressing endoplasmic reticulum stress.展开更多
Spinal cord ischemia-reperfusion injury,a severe form of spinal cord damage,can lead to sensory and motor dysfunction.This injury often occurs after traumatic events,spinal cord surgeries,or thoracoabdominal aortic su...Spinal cord ischemia-reperfusion injury,a severe form of spinal cord damage,can lead to sensory and motor dysfunction.This injury often occurs after traumatic events,spinal cord surgeries,or thoracoabdominal aortic surgeries.The unpredictable nature of this condition,combined with limited treatment options,poses a significant burden on patients,their families,and society.Spinal cord ischemia-reperfusion injury leads to reduced neuronal regenerative capacity and complex pathological processes.In contrast,mitophagy is crucial for degrading damaged mitochondria,thereby supporting neuronal metabolism and energy supply.However,while moderate mitophagy can be beneficial in the context of spinal cord ischemia-reperfusion injury,excessive mitophagy may be detrimental.Therefore,this review aims to investigate the potential mechanisms and regulators of mitophagy involved in the pathological processes of spinal cord ischemia-reperfusion injury.The goal is to provide a comprehensive understanding of recent advancements in mitophagy related to spinal cord ischemia-reperfusion injury and clarify its potential clinical applications.展开更多
The cGAS–STING pathway plays an important role in ischemia-reperfusion injury in the heart,liver,brain,and kidney,but its role and mechanisms in cerebral ischemia-reperfusion injury have not been systematically revie...The cGAS–STING pathway plays an important role in ischemia-reperfusion injury in the heart,liver,brain,and kidney,but its role and mechanisms in cerebral ischemia-reperfusion injury have not been systematically reviewed.Here,we outline the components of the cGAS–STING pathway and then analyze its role in autophagy,ferroptosis,cellular pyroptosis,disequilibrium of calcium homeostasis,inflammatory responses,disruption of the blood–brain barrier,microglia transformation,and complement system activation following cerebral ischemia-reperfusion injury.We further analyze the value of cGAS–STING pathway inhibitors in the treatment of cerebral ischemia-reperfusion injury and conclude that the pathway can regulate cerebral ischemia-reperfusion injury through multiple mechanisms.Inhibition of the cGAS–STING pathway may be helpful in the treatment of cerebral ischemia-reperfusion injury.展开更多
Intestinal ischemia-reperfusion injury(IIRI)is a complex and severe pathophysiological process characterized by oxidative stress,inflammation,and apoptosis.In recent years,the critical roles of extracellular matrix(EC...Intestinal ischemia-reperfusion injury(IIRI)is a complex and severe pathophysiological process characterized by oxidative stress,inflammation,and apoptosis.In recent years,the critical roles of extracellular matrix(ECM)genes and microRNAs(miRNAs)in IIRI have garnered widespread attention.This review aims to systematically summarize the diagnostic and therapeutic potential of ECM gene sets and miRNA regulatory networks in IIRI.First,we review the molecular mechanisms of IIRI,focusing on the dual role of the ECM in tissue injury and repair processes.The expression changes and functions of ECM components such as collagen,elastin,and matrix metalloproteinases during IIRI progression are deeply analyzed.Second,we systematically summarize the regulatory roles of miRNAs in IIRI,particularly the mechanisms and functions of miRNAs such as miR-125b and miR-200a in regulating inflammation,apoptosis,and ECM remodeling.Additionally,this review discusses potential diagnostic biomarkers and treatment strategies based on ECM genes and miRNAs.We extensively evaluate the prospects of miRNA-targeted therapy and ECM component modulation in preventing and treating IIRI,emphasizing the clinical translational potential of these emerging therapies.In conclusion,the diagnostic and therapeutic potential of ECM gene sets and miRNA regulatory networks in IIRI provides new directions for further research,necessitating additional clinical and basic studies to validate and expand these findings for improving clinical outcomes in IIRI patients.展开更多
Background:Although decompression surgery is the optimal treatment for patients with severe degenerative cervical myelopathy(DCM),some individuals experience no improvement or even a decline in neurological function a...Background:Although decompression surgery is the optimal treatment for patients with severe degenerative cervical myelopathy(DCM),some individuals experience no improvement or even a decline in neurological function after surgery,with spinal cord ischemia–reperfusion injury(SCII)identified as the primary cause.Spinal cord compression results in local ischemia and blood perfusion following decompression is fundamental to SCII.However,owing to inadequate perioperative blood flow monitoring,direct evidence regarding the occurrence of SCII after decompression is lacking.The objective of this study was to establish a suitable animal model for investigating the underlying mechanism of spinal cord ischemia–reperfusion injury following decom-pression surgery for degenerative cervical myelopathy(DCM)and to elucidate alterations in neurological function and local blood flow within the spinal cord before and after decompression.Methods:Twenty-four Sprague–Dawley rats were allocated to three groups:the DCM group(cervical compression group,with implanted compression material in the spinal canal,n=8),the DCM-D group(cervical decompression group,with removal of compression material from the spinal canal 4 weeks after implantation,n=8),and the SHAM group(sham operation,n=8).Von Frey test,forepaw grip strength,and gait were assessed within 4 weeks post-implantation.Spinal cord compression was evaluated using magnetic resonance imaging.Local blood flow in the spinal cord was monitored during the perioperative decompression.The rats were sacrificed 1 week after decompression to observe morphological changes in the compressed or decompressed segments of the spinal cord.Additionally,NeuN expression and the oxidative damage marker 8-oxoG DNA were analyzed.Results:Following spinal cord compression,abnormal mechanical pain worsened,and a decrease in forepaw grip strength was observed within 1–4 weeks.Upon decompression,the abnormal mechanical pain subsided,and forepaw grip strength was restored;however,neither reached the level of the sham operation group.Decompression leads to an increase in the local blood flow,indicating improved perfusion of the spinal cord.The number of NeuN-positive cells in the spinal cord of rats in the DCM-D group exceeded that in the DCM group but remained lower than that in the SHAM group.Notably,a higher level of 8-oxoG DNA expression was observed,suggesting oxidative stress following spinal cord decompression.Conclusion:This model is deemed suitable for analyzing the underlying mechanism of SCII following decompressive cervical laminectomy,as we posit that the obtained results are comparable to the clinical progression of degenerative cervical myelopathy(DCM)post-decompression and exhibit analogous neurological alterations.Notably,this model revealed ischemic reperfusion in the spinal cord after decompression,concomitant with oxidative damage,which plausibly underlies the neurological deterioration observed after decompression.展开更多
BACKGROUND Hepatic ischemia-reperfusion injury(HIRI)remains one of the major causes of postoperative liver dysfunction following extensive hepatectomy and liver transplantation.Owing to its progressive and dynamic nat...BACKGROUND Hepatic ischemia-reperfusion injury(HIRI)remains one of the major causes of postoperative liver dysfunction following extensive hepatectomy and liver transplantation.Owing to its progressive and dynamic nature,HIRI may lead to multiple organ failure and a worsened outcome.Treprostinil is a relatively new synthetic prostacyclin analog with a potential beneficial effect against HIRI.Ischemic preconditioning(IP)is a promising method to protect against HIRI.AIM To investigate HIRI biomarkers,their effects on liver and heart,and the effects of treprostinil and IP on these processes.METHODS Forty male Wistar albino rats aged 3-4 months were randomly assigned to four groups of ten,subjected to a 3-hour surgical intervention,and then sacrificed.Hepatic ischemia was induced by clamping the hepatoduodenal ligament for 30 minutes,followed by reperfusion for 120 minutes.Treprostinil(100 ng/kg/minute for 24 hours)or IP before HIRI,no protection,and a sham operation were applied accordingly in each group.Liver and heart histopathology and specific serum and hepatic tissue biomarkers were assessed.RESULTS HIRI deteriorated hepatocellular function and exacerbated liver and myocardial damage in the control group.Furthermore,HIRI triggered cytokine overexpression and protein carbonyl content(P<0.001).Compared with those in the HIRI group,lower troponin I,tumor necrosis factor-α,endothelin-1,and interleukin-1βin serum and liver tissue were significantly correlated with reduced cellular necrosis and improved hepatocellular function in the treprostinil group(P<0.001).Similar but less pronounced effects were observed in the IP group.Both treprostinil and IP had protective effects in hepatic and cardiac tissues.However,treprostinil showed slightly superior cardioprotective efficacy,as evidenced by a statistically significant difference in troponin I levels(P<0.05)and histopathological scoring of myocardium samples,but there were no differences in the other parameters.CONCLUSION HIRI results in oxidative stress and cytokine overexpression,which deteriorate hepatic function and accelerates myocardial damage.Treprostinil and IP are promising strategies for preventing reperfusion-induced cellular and systemic damage.展开更多
Hepatic ischemia-reperfusion injury(IRI)is a prevalent pathophysiological phenomenon encountered during liver surgeries and transplantation,leading to hepatocyte damage and liver dysfunction,which significantly affect...Hepatic ischemia-reperfusion injury(IRI)is a prevalent pathophysiological phenomenon encountered during liver surgeries and transplantation,leading to hepatocyte damage and liver dysfunction,which significantly affects patient prognosis.In recent years,the role of mitophagy in hepatic IRI has garnered considerable attention.Mitochondria,known as the“powerhouses”of the cell,are crucial for maintaining normal cellular physiological functions.During the ischemia-reperfusion process,mitochondria are susceptible to damage,generating excessive harmful substances,such as reactive oxygen species(ROS),which further exacerbate cellular injury.Mitophagy is a selective cellular self-protection mechanism that maintains the quality and quantity balance of mitochondria within cells by clearing damaged or dysfunctional mitochondria.In the context of liver IRI,the activation of mitophagy is of significant importance.On one hand,mitophagy can rapidly remove damaged mitochondria,thereby reducing the release of harmful products and alleviating oxidative stress and cellular damage.Research has indicated that under ischemia-reperfusion conditions,mitophagy-related pathways are activated,promoting the clearance of damaged mitochondria.On the other hand,mitophagy also regulates cellular energy metabolism,providing essential energy support for cells under stress.With the continuous advancement of research,the understanding of the role of mitophagy in hepatic IRI has become increasingly clear.Numerous studies are dedicated to exploring the specific molecular mechanisms of mitophagy and its regulation,aiming to develop new therapeutic strategies to alleviate hepatic IRI.Although studies have demonstrated that mitophagy has a protective effect in hepatic ischemia-reperfusion injury,many issues still require further investigation.First,it is essential to further elucidate the mechanisms underlying the role of mitophagy in ischemia-reperfusion.Additionally,understanding how to mitigate liver ischemia-reperfusion injury through the modulation of mitophagy represents a key focus for future research.Future studies may encompass drug development,gene therapy,and cell therapy approaches aimed at improving the prognosis of patients affected by liver ischemia-reperfusion.展开更多
ABSTRACT:Background:After ischemic stroke,neutrophils hyperactivate,increasing in number and worsening inflammation,causing neural damage.Prior scRNA-seq showed Lrg1 modulates cells subsentence to cerebral ischemiarep...ABSTRACT:Background:After ischemic stroke,neutrophils hyperactivate,increasing in number and worsening inflammation,causing neural damage.Prior scRNA-seq showed Lrg1 modulates cells subsentence to cerebral ischemiareperfusion injury,but its mechanism in regulating neutrophil accumulation/differentiation post-injury is unclear.Methods:Lrg1 knockout impact on neutrophil accumulation was assessed via immunofluorescence and western blot.Three-dimensional reconstruction of immunofluorescent staining analyzed cell-cell interactions among neutrophils and microglia.scRNA-seq of WT and Lrg1^(-/-)mice from GSE245386 and GSE279462 was conducted.Each group conducted oxidative phosphorylation scoring via Gene Set Enrichment Analysis(GSEA),while Metascape was employed to perform GO and KEGG enrichment analyses for elucidating functional mechanisms.CellChat exhibited cell-cell communication.Furthermore,alterations in microglial phagocytic activity were evaluated by immunostaining for CD68,a well-established marker of phagolysosomal activity in phagocytic cells.Brain energy metabolism was evaluated via glutamate dehydrogenase activity and ATP levels with ELISA,and enzyme expression was analyzed by immunofluorescence and western blot.Results:Lrg1 knockout decreased neutrophil accumulation and NET formation in mice.3D immunofluorescence reconstruction confirmed neutrophil co-localization with endothelial cells/microglia.scRNA-seq revealed that the oxidative phosphorylation score was significantly higher in the MCAO/R+WT group compared to both the Sham-operated+WT and Lrg1^(-/-)groups.Notably,the oxidative phosphorylation score was further elevated in the MCAO/R+Lrg1^(-/-)group.Immunostaining showed that Lrg1 knockout elevated CD68+lysosome expression post-MCAO/R,with TMEM119 colocalizing with these lysosomes.MCAO/R raised CD68 expression in ischemic brains,an effect further intensified by Lrg1 knockout.KEGG analysis linked differential genes to oxidative phosphorylation pathways.Validation in MCAO/R vs.sham groups revealed increased ROS production and reduced expression of complex enzymes I-V(NDUFB8,SDHB,UQCRC1,MTCO2,ATP5A1).Lrg1 intervention increased enzyme expression.Immunofluorescence and western blot in brain tissue showed similar patterns in microglia and enzymes I-V.Conclusions:Lrg1 knockout significantly enhances microglial phagocytic activity towards neutrophils subsequent to cerebral ischemia-reperfusion injury,through its regulatory effect on the oxidative phosphorylation pathway.This finding accentuates Lrg1 as a highly potential therapeutic target for intervening in and modulating post-ischemic inflammatory responses.展开更多
Ischemia-reperfusion injury(IRI)remains an unavoidable challenge in liver surgery,with macrophages playing a critical role in its pathogenesis.However,the mechanisms by which macrophages regulate the pathogenesis of I...Ischemia-reperfusion injury(IRI)remains an unavoidable challenge in liver surgery,with macrophages playing a critical role in its pathogenesis.However,the mechanisms by which macrophages regulate the pathogenesis of IRI are not well understood.Through a target-guided screening approach,we identified a small 3 k Da peptide(Sj DX5-271)from various schistosome egg-derived peptides that induced M2 macrophage polarization.SjDX5-271 treatment protected mice against liver IRI by promoting M2 macrophage polarization,and this protective effect was abrogated when the macrophages were depleted.Transcriptomic sequencing showed that the TLR signaling pathway was significantly inhibited in macrophages from the SjDX5-271 treatment group.We further identified that SjDX5-271 promoted M2 macrophage polarization by inhibiting the TLR4/My D88/NF-κB signaling pathway and alleviated hepatic inflammation in liver IRI.Collectively,SjDX5-271 exhibited some promising therapeutic effects in IRI and represented a novel therapeutic approach,potentially applicable to other immune-related diseases.The current study demonstrates the potential of new biologics from the parasite,enhances our understanding of host-parasite interplay,and provides a blueprint for future therapies for immunerelated diseases.展开更多
BACKGROUND:Hepatic ischemia-reperfusion(I/R)injury is a major challenge in liver surgery and transplantation.Bromodomain protein 4(BRD4)has emerged as a promising target due to its role in oxidative stress and inflamm...BACKGROUND:Hepatic ischemia-reperfusion(I/R)injury is a major challenge in liver surgery and transplantation.Bromodomain protein 4(BRD4)has emerged as a promising target due to its role in oxidative stress and inflammation.JQ-1,a specific BRD4 inhibitor,has shown protective effects on organs suffering I/R injury.This study aims to investigate the expression of BRD4 in liver tissues after I/R injury and to explore its role in this process using JQ-1 both in vivo and in vitro.METHODS:Our study established a mouse model of hepatic I/R injury and investigated the protective effect of JQ-1.We compared the histological features,BRD4 expression,and liver enzyme levels between JQ-1-treated and untreated groups.Additionally,the antioxidant properties of JQ-1 were analyzed in RAW 264.7 cells by evaluating cytokine expression,NLRP3 inflammasome activity,and reactive oxygen species production.RESULTS:BRD4 was abundantly expressed in liver tissues after hepatic I/R injury,while JQ-1 treatment had antioxidant and hepatoprotective effects.JQ-1 also suppressed pro-inflammatory cytokine release in vitro.Furthermore,we clarified the mechanism by which JQ-1 enhances liver injury recovery through Kupffer cells by blocking the NOD-like receptor thermal protein domain-associated protein 3(NLRP3)/caspase-1 pathway.CONCLUSION:JQ-1 has potential as a pre-clinical emergency therapy for hepatic I/R injury.Its ability to inhibit BRD4 and modulate the inflammatory response in Kupffer cells offers a promising avenue for future clinical intervention.展开更多
With the wide application of thrombolytic drugs and the advancement of endovascular therapeutic techniques, the recanalization treatment of acute artery occlusion in ischemic stroke (IS) has made a leap forward, but i...With the wide application of thrombolytic drugs and the advancement of endovascular therapeutic techniques, the recanalization treatment of acute artery occlusion in ischemic stroke (IS) has made a leap forward, but ischemic brain tissues still face ischemia-reperfusion injury after recanalization. Nowadays, effective neurological protective agents still cannot completely resist the multiple damages of ischemia-reperfusion injury. As an iron-dependent mode of programmed cell death, ferroptosis occupies an important position in ischemia-reperfusion injury. Selenium plays a unique protective role in ischemia-reperfusion injury as an active site element in the center of glutathione peroxidase. Therefore, the study mainly aims to review the protective role of selenium in IS and the related mechanisms, as well as the effect of selenium on the risk factors of IS.展开更多
AIM:To explore the immune cell infiltration and molecular mechanisms of retinal ischemia-reperfusion injury(RIRI)to identify potential therapeutic targets.METHODS:In the bulk RNA-seq analysis,This study performed diff...AIM:To explore the immune cell infiltration and molecular mechanisms of retinal ischemia-reperfusion injury(RIRI)to identify potential therapeutic targets.METHODS:In the bulk RNA-seq analysis,This study performed differential gene expression analysis,weighted gene co-expression network analysis,and protein-protein interaction network analysis to identify hub genes.QuanTIseq was used to determine the composition of infiltrating immune cells.Following the identification of hub genes,single-cell RNA-seq analysis was employed to pinpoint the specific immune cell types expressing these hub genes.Cell-cell communication analysis to explore signaling pathways and interactions between immune cells was further performed.Finally,the expression of these key immune regulators in vivo using quantitative real-time polymerase chain reaction(qRT-PCR)was validated.RESULTS:Bulk RNA-seq analysis identified Stat2,Irf7,Irgm1,Igtp,Parp9,Irgm2,Nlrc5,and Tap1 as hub genes,with strong correlations to immune cell infiltration.Single-cell RNA-seq analysis further revealed six immune cell clusters,showing Irf7 predominantly in microglia and Tap1 in dendritic cells(DCs).And cell-cell communication analysis showed that microglia and DCs play central roles in coordinating immune activity.qRT-PCR validated the upregulation of these genes.CONCLUSION:In the acute phase of RIRI,Irf7 and Tap1 may be the potential therapeutic targets to reduce inflammation and promote neurological function recovery.展开更多
BACKGROUND Ferroptosis is a newly recognized form of regulated cell death characterized by iron-dependent accumulation of lipid reactive oxygen species.It has been extensively studied in various diseases,including can...BACKGROUND Ferroptosis is a newly recognized form of regulated cell death characterized by iron-dependent accumulation of lipid reactive oxygen species.It has been extensively studied in various diseases,including cancer,Parkinson’s disease,and stroke.However,its precise role and underlying mechanisms in ischemia/reperfusion injury,particularly in the intestinal ischemia-reperfusion(IIR),remain unclear.In current work,we aimed to investigate the participation of histone lactylation during IIR progression.AIM To investigate the role of mitochondrial alanyl-tRNA synthetase 2(AARS2)in ferroptosis and its epigenetic regulation of acyl-CoA synthetase long-chain family member 4(ACSL4)through histone lactylation during IIR injury.METHODS We established a mouse model to mimic IIR and conducted AARS2 knockdown as treatment.The expression of AARS2 in intestinal tissues was measured by western blot.The integrity of intestinal tissues was detected by hematoxylin and eosin staining,serum fatty acid-binding protein,protein levels of ZO-1 and occluding.An in vitro hypoxia-reperfusion(H/R)cell model was established,and cell viability was measured by CCK-8.The in vitro and in vivo ferroptosis was determined by the accumulation of Fe2+and malondialdehyde(MDA).The epigenetic regulation of ACSL4 by AARS2 was detected by chromatin immunoprecipitation(ChIP)assay and luciferase reporter assay.RESULTS We observed a notable elevated AARS2 level in intestinal tissue of mice in IIR model group,which was reversed by shAARS2 treatment.Knockdown of AARS2 repressed alleviated intestinal barrier disruption and repressed the accumulation of ferroptosis biomarker Fe2+and MDA during IIR.The in vitro results showed that shAARS2 alleviated impaired cell viability caused by H/R,as well as repressed ferroptosis.Knockdown of AARS2 notably downregulated the RNA and protein expression of ACSL4.Mechanistically,knockdown of AARS2 downregulated the enrichment of H3K18 La modification on AARS2,as well as suppressed its promoter activity.Overexpression of AARS2 could abolish the protective effects of shACSL4 in vitro.CONCLUSION The elevation of AARS2 during IIR led to cell ferroptosis via epigenetically upregulating the expression of ACSL4.Our findings presented AARS2 as a promising therapeutic target for IIR.展开更多
Background:The aim of the study was to explore a feasible method for alleviating limb ischemia-reperfusion injury(LI/RI)through the use of a high-concentration citrate solution(HC-A solution)for limb perfusion(LP).Met...Background:The aim of the study was to explore a feasible method for alleviating limb ischemia-reperfusion injury(LI/RI)through the use of a high-concentration citrate solution(HC-A solution)for limb perfusion(LP).Methods:Eighteen pigs were divided into three groups:the Sham group,LI/RI group,and HCA group.The Sham group underwent exposure of the iliac artery and vein.The LI/RI group underwent tourniquet placement and clamping of the iliac artery and vein to simulate LI/RI.The HCA group received HC-A solution LP for 30 min through the left iliac artery below the level of blood flow occlusion based on the LI/RI group.Oxidative stress markers and inflammatory response markers were compared among the three groups.Results:Compared to the LI/RI group,the HCA group showed significantly lower levels of serum creatine kinase(CK),lactate dehydrogenase(LDH),malondialdehyde(MDA),tumor necrosis factor-α(TNF-α),aspartate aminotransferase(AST),and ala-nine aminotransferase(ALT),and significantly greater activities of serum superoxide dismutase(SOD)(p<0.05).There were no significant differences in serum interleukin-6(IL-6)or in muscle MDA,SOD,TNF-α,and IL-6 between the HCA group and the LI/RI group(p>0.05).Compared to the LI/RI group,MDA,TNF-α,and IL-6 levels in the liver were significantly lower in the HCA group(p<0.05),while SOD activities were not significantly different(p>0.05).Histopathological examination revealed reduced skeletal muscle and liver damage in the HCA group compared to the LI/RI group.Conclusions:HC-A solution LP can alleviate liver damage caused by LI/RI in pigs.展开更多
BACKGROUND Hepatic ischemia-reperfusion(I/R)injury related to liver transplantation and hepatic resection remains a challenge in clinical practice.Accumulating evidence indicates that mitochondrial dysfunction is a cr...BACKGROUND Hepatic ischemia-reperfusion(I/R)injury related to liver transplantation and hepatic resection remains a challenge in clinical practice.Accumulating evidence indicates that mitochondrial dysfunction is a critical cause of I/R injury.The protein 4-nitrophenylphosphatase domain and non-neuronal SNAP25-like protein homolog 1(NIPSNAP1)is involved in the regulation of mitophagy and the recruitment of autophagy receptor proteins independent of PTEN induced putative kinase 1.AIM To clarify the protective mechanism of NIPSNAP1 against hepatic I/R,with a focus on mitophagy and mitochondrial dynamics,as well as the potential mechanism by which n6-methyladenosine(m6A)modification regulates NIPSNAP1.METHODS Mice were administered an adeno-associated virus in vivo and a hepatic I/R model was established via portal vein interruption followed by reperfusion to explore the effect of NIPSNAP1 on hepatic I/R.HepG2 cells were subjected to hypoxia/reoxygenation treatment in vitro.RESULTS We observed a significant downregulation of both NIPSNAP1 and insulin-like growth factor 2 mRNA-binding protein 2(IGF2BP2)expression in vivo and in vitro.NIPSNAP1 knockdown impaired mitophagy and disrupted mitochondrial dynamics;in contrast,NIPSNAP1 overexpression resulted in the opposite effects.Further studies revealed that IGF2BP2 functions as an m6A reader that targets and binds NIPSNAP1,thereby regulating its mRNA stability.CONCLUSION NIPSNAP1 prevents hepatic I/R injury by promoting mitophagy and maintaining mitochondrial homeostasis,serving as a novel target of the m6A reader IGF2BP2.Therefore,targeting the IGF2BP2/NIPSNAP1 axis may facilitate the development of better therapeutics for hepatic I/R.展开更多
PI3K/AKT/mTOR signaling pathway is a key pathway of myocardial ischemia-reperfusion injury(MIRI).The mechanism of action is mainly oxidative stress,inflammatory response,calcium overload,ferroptosis,autophagy,and apop...PI3K/AKT/mTOR signaling pathway is a key pathway of myocardial ischemia-reperfusion injury(MIRI).The mechanism of action is mainly oxidative stress,inflammatory response,calcium overload,ferroptosis,autophagy,and apoptosis.MIRI belongs to the category of chest obstruction in traditional Chinese medicine,and its etiology and pathogenesis are mainly“Yang Wei Yin Xian.”Traditional Chinese medicine has the effect of multi-target and multi-component effect,and has played a significant role in the treatment of MIRI in recent years.At present,the monomers of traditional Chinese medicine mainly include saponins,flavonoids,alkaloids,terpenoids,and phenols,and the compounds mainly include Zhigancao Decoction,Zhenyuan Capsule,Jiawei Shenqibai Powder,Qili Qiangxin Capsule,Tongmai Yangxin Pill,Zhilong Huoxue Tongyu Capsule,Guizhi Tongluo Tablets,etc.This paper reviews the research on the improvement of MIRI by regulating PI3K/AKT/mTOR signaling pathway in recent years,and expounds the mechanism and advantages of traditional Chinese medicine in the treatment of MIRI.展开更多
Objective:To investigate the potential protective effect of Shexiang Tongxin dropping pills(STDP)on ischemia-reperfusion injury and its underlying mechanisms in improving endothelial cell function in coronary microvas...Objective:To investigate the potential protective effect of Shexiang Tongxin dropping pills(STDP)on ischemia-reperfusion injury and its underlying mechanisms in improving endothelial cell function in coronary microvascular disease(CMVD).Methods:A rat model of myocardial ischemia-reperfusion injury with CMVD was established using ligation and reperfusion of the left anterior descending artery.The effect of STDP(21.6 mg/kg)on cardiac function was evaluated using echocardiography,hematoxylin-eosin staining,and Evans blue staining.The effects of STDP on the microvascular endothelial barrier were assessed based on nitric oxide production,endothelial nitric oxide synthase expression,structural variety of tight junctions(TJs),and the expression of zonula occludens-1(ZO-1),claudin-5,occludin,and vascular endothelial(VE)-cadherin proteins.The mechanisms of STDP(50 and 100 ng/mL)were evaluated by examining the expression of sphingosine 1-phosphate receptor 2(S1PR2),Ras Homolog family member A(RhoA),and Rho-associated coiled-coil-containing protein kinase(ROCK)proteins and the distribution of ZO-1,VE-cadherin,and Factin proteins in an oxygen and glucose deprivation/reoxygenation model.Results:The administration of STDP on CMVD rat model significantly improved cardiac and microvascular endothelial cell barrier functions(all P<.05).STDP enhanced the structural integrity of coronary microvascular positioning and distribution by clarifying and completing TJs and increasing the expression of ZO-1,occludin,claudin-5,and VE-cadherin in vivo(all P<.05).The S1PR2/RhoA/ROCK pathway was inhibited by STDP in vitro,leading to the regulation of endothelial cell TJs,adhesion junctions,and cytoskeletal morphology.Conclusion:STDP showed protective effects on cardiac impairment and microvascular endothelial barrier injury in CMVD model rats induced by myocardial ischemia-reperfusion injury through the modulation of the S1PR2/RhoA/ROCK pathway.展开更多
Lung ischemia-reperfusion injury(LIRI),an acute lung injury syndrome triggered by lung transplantation or distal organ ischemia,has long been a difficult and hot issue in clinical research.In this study,we proposed a ...Lung ischemia-reperfusion injury(LIRI),an acute lung injury syndrome triggered by lung transplantation or distal organ ischemia,has long been a difficult and hot issue in clinical research.In this study,we proposed a simple and less invasive reversible LIRI surgical protocol,achieved by improving the mouse left hilar entrapment model,which significantly improved the operability of the experiment and the reproducibility of the results.The protocol achieves precise control of the ischemic and reperfusion processes by visualizing transoral intubation,using reversible ligation of live knots to clamp the left hilar,and temporary closure of the thoracic cavity during ischemia.The reversible survival model we constructed not only provides a reliable tool to study the cellular and molecular mechanisms of LIRI but also can be used to assess the stage of injury regression,experimental pneumonia,and survival in mice.In addition,it simplifies the lung portal separation clamping operation for reversible clamping and provides an easy-to-learn visual tracheal intubation method that can be quickly mastered and replicated by beginners for consistent and reliable results.展开更多
AIM To study the effects of warm ischemia-reperfusion(I/R) injury on hepatic morphology at the ultrastructural level and to analyze the expression of the thioredoxin(TRX)and glutaredoxin(GRX) systems.METHODS Eleven pa...AIM To study the effects of warm ischemia-reperfusion(I/R) injury on hepatic morphology at the ultrastructural level and to analyze the expression of the thioredoxin(TRX)and glutaredoxin(GRX) systems.METHODS Eleven patients undergoing liver resection were subjected to portal triad clamping(PTC). Liver biopsies were collected at three time points; first prior to PTC(baseline), 20 min after PTC(post-ischemia) and 20 min after reperfusion(post-reperfusion). Electron microscopy and morphometry were used to study and quantify ultrastructural changes, respectively. Additionally, gene expression analysis of TRX and GRX isoforms was performed by quantitative PCR. For further validation of redox protein status, immunogold staining was performed for the isoforms GRX1 and TRX1.RESULTS Post-ischemia, a significant loss of the liver sinusoidal endothelial cell(LSEC) lining was observed(P = 0.0003) accompanied by a decrease of hepatocyte microvilli in the space of Disse. Hepatocellular morphology was well preserved apart from the appearance of crystalline mitochondrial inclusions in 7 out of 11 patients. Postreperfusion biopsies had similar features as post-ischemia with the exception of signs of a reactivation of the LSECs. No changes in the expression of redox-regulatory genes could be observed at mR NA level of the isoforms of the TRX family but immunoelectron microscopy indicated a redistribution of TRX1 within the cell.CONCLUSION At the ultrastructural level, the major impact of hepatic warm I/R injury after PTC was borne by the LSECs with detachment and reactivation at ischemia and reperfusion, respectively. Hepatocytes morphology were well preserved. Crystalline inclusions in mitochondria were observed in the hepatocyte after ischemia.展开更多
Background Myocardial ischemia/reperfusion(I/R)injury remains a great challenge in clinical therapy.Recent studies indicated that circularRNA(circRNA)play an important role in the regulation of atherosclerotic heart d...Background Myocardial ischemia/reperfusion(I/R)injury remains a great challenge in clinical therapy.Recent studies indicated that circularRNA(circRNA)play an important role in the regulation of atherosclerotic heart disease.However,as a new circRNA,the effect of circRNA regulating ischemia-reperfusion through microRNA-1(CRIRTM)on myocardial apoptosis during myocardial ischemia/reperfusion is still unknown.The purpose of this study was to investigate the effect of CRIRTM on I/R-induced apoptosis and the expression of microRNA-1(miR-1)and calmodulin(CaM)in cultured neonatal rat ventricular cardiomyocytes(NRVCs)in vitro and SD rat hypoxia-reoxygenation model in vivo.Methods NRVCs were subjected to hypoxia for 30 minutes and then oxygen was added for 12 hours to establish the model of cell ischemia reperfusion.The SD rats were subjected to the surgery of ligating the anterior descending branch of the left coronary artery for 4 hours and then reperfused for 48 hours to establish myocardial ischemia animal model.The expression of CRIRTM,miIR-1 and CaM was detected by quantitative real-time polymerase chain reaction(qRT-PCR).TUNEL was used to analyze the apoptosis level of cells and animal myocardium after small interfering RNA(siRNA)down-regulation of CRIRTM.The expression of CRIRTM in the blood of patients with myocardial infarction was detected,and the downstream of CRIRTM was analyzed by Target Scan.Results The specific upregulation of CRIRTM was detected in the plasma of patients with ST-segment elevation myocardial infarction after reperfusion,and multiple binding sites between CRIRTM and miR-1 were found by Target Scan analysis.The expression of CRIRTM and CaM was up-regulated and the expression of miR-1 was down-regulated in cells and rats I/R model.Both in vivo and in vitro,cardiomyocyte apoptosis induced by myocardial ischemia-reperfusion will increase after down-regulation of CRIRTM.It may be due to the decreased inhibition of miR-1 by down-regulated CRIRTM,which increases the degradation of CaM by miR-1 and aggravates cardiomyocyte apoptosis.Conclusions Our data demonstrated that CRIRTM alleviates myocardial ischemia-reperfusion injury via suppressing miR-1 leading to enhanced CaM levels.[S Chin J Cardiol 2021;22(1):30-37]展开更多
基金supported in part by the National Key Research&Development Program of China,No.2022YFA1104900(to LS)the National Natural Science Foundation of China,Nos.82371175,82071535(both to LS),82101614(to YP)+5 种基金the International Science and Technology Cooperation Projects of Guangdong Province,No.2023A0505050121(to LS)Guangdong Basic and Applied Basic Research Foundation,Nos.2022B1515130007(to LS),2023A1515030012(to SZ),2022A1515010666(to WL)the Science and Technology Program of Guangzhou,Nos.202102070001(to LS),202201010041(to YP)Shenzhen Basic Research Grant,Nos.JCYJ20200109140414636,JCYJ20230807145103007(both to WL)awarded a Royal Society Newton Advanced Fellowship,No.AOMS-NAF0051003in collaboration with Zoltán Molnár,Department of Physiology,Anatomy and Genetics,University of Oxford(2017–2021)。
文摘Neuroserpin,a secreted protein that belongs to the serpin superfamily of serine protease inhibitors,is highly expressed in the central nervous system and plays multiple roles in brain development and pathology.As a natural inhibitor of recombinant tissue plasminogen activator,neuroserpin inhibits the increased activity of tissue plasminogen activator in ischemic conditions and extends the therapeutic windows of tissue plasminogen activator for brain ischemia.However,the neuroprotective mechanism of neuroserpin against ischemic stroke remains unclear.In this study,we used a mouse model of middle cerebral artery occlusion and oxygen-glucose deprivation/reperfusion-injured cortical neurons as in vivo and in vitro ischemia-reperfusion models,respectively.The models were used to investigate the neuroprotective effects of neuroserpin.Our findings revealed that endoplasmic reticulum stress was promptly triggered following ischemia,initially manifesting as the acute activation of endoplasmic reticulum stress transmembrane sensors and the suppression of protein synthesis,which was followed by a later apoptotic response.Notably,ischemic stroke markedly downregulated the expression of neuroserpin in cortical neurons.Exogenous neuroserpin reversed the activation of multiple endoplasmic reticulum stress signaling molecules,the reduction in protein synthesis,and the upregulation of apoptotic transcription factors.This led to a reduction in neuronal death induced by oxygen/glucose deprivation and reperfusion,as well as decreased cerebral infarction and neurological dysfunction in mice with middle cerebral artery occlusion.However,the neuroprotective effects of neuroserpin were markedly inhibited by endoplasmic reticulum stress activators thapsigargin and tunicamycin.Our findings demonstrate that neuroserpin exerts neuroprotective effects on ischemic stroke by suppressing endoplasmic reticulum stress.
基金supported by Cuiying Scientific and Technological Innovation Program of Second Hospital of Lanzhou University,Nos.CY2023-QN-B18(to YD),2020QN-16(to YZ)the Natural Science Foundation of Gansu Province,No.22JR11RA082(to YZ)Key R&D Plan of Gansu Provincial Department of Science and Technology-Social Development Projects,No.23YFFA0043(to XK).
文摘Spinal cord ischemia-reperfusion injury,a severe form of spinal cord damage,can lead to sensory and motor dysfunction.This injury often occurs after traumatic events,spinal cord surgeries,or thoracoabdominal aortic surgeries.The unpredictable nature of this condition,combined with limited treatment options,poses a significant burden on patients,their families,and society.Spinal cord ischemia-reperfusion injury leads to reduced neuronal regenerative capacity and complex pathological processes.In contrast,mitophagy is crucial for degrading damaged mitochondria,thereby supporting neuronal metabolism and energy supply.However,while moderate mitophagy can be beneficial in the context of spinal cord ischemia-reperfusion injury,excessive mitophagy may be detrimental.Therefore,this review aims to investigate the potential mechanisms and regulators of mitophagy involved in the pathological processes of spinal cord ischemia-reperfusion injury.The goal is to provide a comprehensive understanding of recent advancements in mitophagy related to spinal cord ischemia-reperfusion injury and clarify its potential clinical applications.
基金supported by Yuan Du Scholars,Clinical Research Center of Affiliated Hospital of Shandong Second Medical University,No.2022WYFYLCYJ02Weifang Key Laboratory,Weifang Science and Technology Development Plan Project Medical Category,No.2022YX093.
文摘The cGAS–STING pathway plays an important role in ischemia-reperfusion injury in the heart,liver,brain,and kidney,but its role and mechanisms in cerebral ischemia-reperfusion injury have not been systematically reviewed.Here,we outline the components of the cGAS–STING pathway and then analyze its role in autophagy,ferroptosis,cellular pyroptosis,disequilibrium of calcium homeostasis,inflammatory responses,disruption of the blood–brain barrier,microglia transformation,and complement system activation following cerebral ischemia-reperfusion injury.We further analyze the value of cGAS–STING pathway inhibitors in the treatment of cerebral ischemia-reperfusion injury and conclude that the pathway can regulate cerebral ischemia-reperfusion injury through multiple mechanisms.Inhibition of the cGAS–STING pathway may be helpful in the treatment of cerebral ischemia-reperfusion injury.
基金Supported by Health Science and Technology Programme of Zhejiang Province,No.2022KY1391.
文摘Intestinal ischemia-reperfusion injury(IIRI)is a complex and severe pathophysiological process characterized by oxidative stress,inflammation,and apoptosis.In recent years,the critical roles of extracellular matrix(ECM)genes and microRNAs(miRNAs)in IIRI have garnered widespread attention.This review aims to systematically summarize the diagnostic and therapeutic potential of ECM gene sets and miRNA regulatory networks in IIRI.First,we review the molecular mechanisms of IIRI,focusing on the dual role of the ECM in tissue injury and repair processes.The expression changes and functions of ECM components such as collagen,elastin,and matrix metalloproteinases during IIRI progression are deeply analyzed.Second,we systematically summarize the regulatory roles of miRNAs in IIRI,particularly the mechanisms and functions of miRNAs such as miR-125b and miR-200a in regulating inflammation,apoptosis,and ECM remodeling.Additionally,this review discusses potential diagnostic biomarkers and treatment strategies based on ECM genes and miRNAs.We extensively evaluate the prospects of miRNA-targeted therapy and ECM component modulation in preventing and treating IIRI,emphasizing the clinical translational potential of these emerging therapies.In conclusion,the diagnostic and therapeutic potential of ECM gene sets and miRNA regulatory networks in IIRI provides new directions for further research,necessitating additional clinical and basic studies to validate and expand these findings for improving clinical outcomes in IIRI patients.
基金Wangjing Hospital Affiliated to China Academy of Chinese Medical Sciences,Grant/Award Number:WJYY-YJKT-2022-13。
文摘Background:Although decompression surgery is the optimal treatment for patients with severe degenerative cervical myelopathy(DCM),some individuals experience no improvement or even a decline in neurological function after surgery,with spinal cord ischemia–reperfusion injury(SCII)identified as the primary cause.Spinal cord compression results in local ischemia and blood perfusion following decompression is fundamental to SCII.However,owing to inadequate perioperative blood flow monitoring,direct evidence regarding the occurrence of SCII after decompression is lacking.The objective of this study was to establish a suitable animal model for investigating the underlying mechanism of spinal cord ischemia–reperfusion injury following decom-pression surgery for degenerative cervical myelopathy(DCM)and to elucidate alterations in neurological function and local blood flow within the spinal cord before and after decompression.Methods:Twenty-four Sprague–Dawley rats were allocated to three groups:the DCM group(cervical compression group,with implanted compression material in the spinal canal,n=8),the DCM-D group(cervical decompression group,with removal of compression material from the spinal canal 4 weeks after implantation,n=8),and the SHAM group(sham operation,n=8).Von Frey test,forepaw grip strength,and gait were assessed within 4 weeks post-implantation.Spinal cord compression was evaluated using magnetic resonance imaging.Local blood flow in the spinal cord was monitored during the perioperative decompression.The rats were sacrificed 1 week after decompression to observe morphological changes in the compressed or decompressed segments of the spinal cord.Additionally,NeuN expression and the oxidative damage marker 8-oxoG DNA were analyzed.Results:Following spinal cord compression,abnormal mechanical pain worsened,and a decrease in forepaw grip strength was observed within 1–4 weeks.Upon decompression,the abnormal mechanical pain subsided,and forepaw grip strength was restored;however,neither reached the level of the sham operation group.Decompression leads to an increase in the local blood flow,indicating improved perfusion of the spinal cord.The number of NeuN-positive cells in the spinal cord of rats in the DCM-D group exceeded that in the DCM group but remained lower than that in the SHAM group.Notably,a higher level of 8-oxoG DNA expression was observed,suggesting oxidative stress following spinal cord decompression.Conclusion:This model is deemed suitable for analyzing the underlying mechanism of SCII following decompressive cervical laminectomy,as we posit that the obtained results are comparable to the clinical progression of degenerative cervical myelopathy(DCM)post-decompression and exhibit analogous neurological alterations.Notably,this model revealed ischemic reperfusion in the spinal cord after decompression,concomitant with oxidative damage,which plausibly underlies the neurological deterioration observed after decompression.
文摘BACKGROUND Hepatic ischemia-reperfusion injury(HIRI)remains one of the major causes of postoperative liver dysfunction following extensive hepatectomy and liver transplantation.Owing to its progressive and dynamic nature,HIRI may lead to multiple organ failure and a worsened outcome.Treprostinil is a relatively new synthetic prostacyclin analog with a potential beneficial effect against HIRI.Ischemic preconditioning(IP)is a promising method to protect against HIRI.AIM To investigate HIRI biomarkers,their effects on liver and heart,and the effects of treprostinil and IP on these processes.METHODS Forty male Wistar albino rats aged 3-4 months were randomly assigned to four groups of ten,subjected to a 3-hour surgical intervention,and then sacrificed.Hepatic ischemia was induced by clamping the hepatoduodenal ligament for 30 minutes,followed by reperfusion for 120 minutes.Treprostinil(100 ng/kg/minute for 24 hours)or IP before HIRI,no protection,and a sham operation were applied accordingly in each group.Liver and heart histopathology and specific serum and hepatic tissue biomarkers were assessed.RESULTS HIRI deteriorated hepatocellular function and exacerbated liver and myocardial damage in the control group.Furthermore,HIRI triggered cytokine overexpression and protein carbonyl content(P<0.001).Compared with those in the HIRI group,lower troponin I,tumor necrosis factor-α,endothelin-1,and interleukin-1βin serum and liver tissue were significantly correlated with reduced cellular necrosis and improved hepatocellular function in the treprostinil group(P<0.001).Similar but less pronounced effects were observed in the IP group.Both treprostinil and IP had protective effects in hepatic and cardiac tissues.However,treprostinil showed slightly superior cardioprotective efficacy,as evidenced by a statistically significant difference in troponin I levels(P<0.05)and histopathological scoring of myocardium samples,but there were no differences in the other parameters.CONCLUSION HIRI results in oxidative stress and cytokine overexpression,which deteriorate hepatic function and accelerates myocardial damage.Treprostinil and IP are promising strategies for preventing reperfusion-induced cellular and systemic damage.
文摘Hepatic ischemia-reperfusion injury(IRI)is a prevalent pathophysiological phenomenon encountered during liver surgeries and transplantation,leading to hepatocyte damage and liver dysfunction,which significantly affects patient prognosis.In recent years,the role of mitophagy in hepatic IRI has garnered considerable attention.Mitochondria,known as the“powerhouses”of the cell,are crucial for maintaining normal cellular physiological functions.During the ischemia-reperfusion process,mitochondria are susceptible to damage,generating excessive harmful substances,such as reactive oxygen species(ROS),which further exacerbate cellular injury.Mitophagy is a selective cellular self-protection mechanism that maintains the quality and quantity balance of mitochondria within cells by clearing damaged or dysfunctional mitochondria.In the context of liver IRI,the activation of mitophagy is of significant importance.On one hand,mitophagy can rapidly remove damaged mitochondria,thereby reducing the release of harmful products and alleviating oxidative stress and cellular damage.Research has indicated that under ischemia-reperfusion conditions,mitophagy-related pathways are activated,promoting the clearance of damaged mitochondria.On the other hand,mitophagy also regulates cellular energy metabolism,providing essential energy support for cells under stress.With the continuous advancement of research,the understanding of the role of mitophagy in hepatic IRI has become increasingly clear.Numerous studies are dedicated to exploring the specific molecular mechanisms of mitophagy and its regulation,aiming to develop new therapeutic strategies to alleviate hepatic IRI.Although studies have demonstrated that mitophagy has a protective effect in hepatic ischemia-reperfusion injury,many issues still require further investigation.First,it is essential to further elucidate the mechanisms underlying the role of mitophagy in ischemia-reperfusion.Additionally,understanding how to mitigate liver ischemia-reperfusion injury through the modulation of mitophagy represents a key focus for future research.Future studies may encompass drug development,gene therapy,and cell therapy approaches aimed at improving the prognosis of patients affected by liver ischemia-reperfusion.
基金supported by the Foundation Project:National Natural Science Foundation of China(Nos.:82460249,82100417,81760094)The Foundation of Jiangxi Provincial Department of Science and Technology Outstanding Youth Fund Project(20242BAB23080).
文摘ABSTRACT:Background:After ischemic stroke,neutrophils hyperactivate,increasing in number and worsening inflammation,causing neural damage.Prior scRNA-seq showed Lrg1 modulates cells subsentence to cerebral ischemiareperfusion injury,but its mechanism in regulating neutrophil accumulation/differentiation post-injury is unclear.Methods:Lrg1 knockout impact on neutrophil accumulation was assessed via immunofluorescence and western blot.Three-dimensional reconstruction of immunofluorescent staining analyzed cell-cell interactions among neutrophils and microglia.scRNA-seq of WT and Lrg1^(-/-)mice from GSE245386 and GSE279462 was conducted.Each group conducted oxidative phosphorylation scoring via Gene Set Enrichment Analysis(GSEA),while Metascape was employed to perform GO and KEGG enrichment analyses for elucidating functional mechanisms.CellChat exhibited cell-cell communication.Furthermore,alterations in microglial phagocytic activity were evaluated by immunostaining for CD68,a well-established marker of phagolysosomal activity in phagocytic cells.Brain energy metabolism was evaluated via glutamate dehydrogenase activity and ATP levels with ELISA,and enzyme expression was analyzed by immunofluorescence and western blot.Results:Lrg1 knockout decreased neutrophil accumulation and NET formation in mice.3D immunofluorescence reconstruction confirmed neutrophil co-localization with endothelial cells/microglia.scRNA-seq revealed that the oxidative phosphorylation score was significantly higher in the MCAO/R+WT group compared to both the Sham-operated+WT and Lrg1^(-/-)groups.Notably,the oxidative phosphorylation score was further elevated in the MCAO/R+Lrg1^(-/-)group.Immunostaining showed that Lrg1 knockout elevated CD68+lysosome expression post-MCAO/R,with TMEM119 colocalizing with these lysosomes.MCAO/R raised CD68 expression in ischemic brains,an effect further intensified by Lrg1 knockout.KEGG analysis linked differential genes to oxidative phosphorylation pathways.Validation in MCAO/R vs.sham groups revealed increased ROS production and reduced expression of complex enzymes I-V(NDUFB8,SDHB,UQCRC1,MTCO2,ATP5A1).Lrg1 intervention increased enzyme expression.Immunofluorescence and western blot in brain tissue showed similar patterns in microglia and enzymes I-V.Conclusions:Lrg1 knockout significantly enhances microglial phagocytic activity towards neutrophils subsequent to cerebral ischemia-reperfusion injury,through its regulatory effect on the oxidative phosphorylation pathway.This finding accentuates Lrg1 as a highly potential therapeutic target for intervening in and modulating post-ischemic inflammatory responses.
基金supported by the National Natural Science Foundation of China(Grant No.81971965)。
文摘Ischemia-reperfusion injury(IRI)remains an unavoidable challenge in liver surgery,with macrophages playing a critical role in its pathogenesis.However,the mechanisms by which macrophages regulate the pathogenesis of IRI are not well understood.Through a target-guided screening approach,we identified a small 3 k Da peptide(Sj DX5-271)from various schistosome egg-derived peptides that induced M2 macrophage polarization.SjDX5-271 treatment protected mice against liver IRI by promoting M2 macrophage polarization,and this protective effect was abrogated when the macrophages were depleted.Transcriptomic sequencing showed that the TLR signaling pathway was significantly inhibited in macrophages from the SjDX5-271 treatment group.We further identified that SjDX5-271 promoted M2 macrophage polarization by inhibiting the TLR4/My D88/NF-κB signaling pathway and alleviated hepatic inflammation in liver IRI.Collectively,SjDX5-271 exhibited some promising therapeutic effects in IRI and represented a novel therapeutic approach,potentially applicable to other immune-related diseases.The current study demonstrates the potential of new biologics from the parasite,enhances our understanding of host-parasite interplay,and provides a blueprint for future therapies for immunerelated diseases.
文摘BACKGROUND:Hepatic ischemia-reperfusion(I/R)injury is a major challenge in liver surgery and transplantation.Bromodomain protein 4(BRD4)has emerged as a promising target due to its role in oxidative stress and inflammation.JQ-1,a specific BRD4 inhibitor,has shown protective effects on organs suffering I/R injury.This study aims to investigate the expression of BRD4 in liver tissues after I/R injury and to explore its role in this process using JQ-1 both in vivo and in vitro.METHODS:Our study established a mouse model of hepatic I/R injury and investigated the protective effect of JQ-1.We compared the histological features,BRD4 expression,and liver enzyme levels between JQ-1-treated and untreated groups.Additionally,the antioxidant properties of JQ-1 were analyzed in RAW 264.7 cells by evaluating cytokine expression,NLRP3 inflammasome activity,and reactive oxygen species production.RESULTS:BRD4 was abundantly expressed in liver tissues after hepatic I/R injury,while JQ-1 treatment had antioxidant and hepatoprotective effects.JQ-1 also suppressed pro-inflammatory cytokine release in vitro.Furthermore,we clarified the mechanism by which JQ-1 enhances liver injury recovery through Kupffer cells by blocking the NOD-like receptor thermal protein domain-associated protein 3(NLRP3)/caspase-1 pathway.CONCLUSION:JQ-1 has potential as a pre-clinical emergency therapy for hepatic I/R injury.Its ability to inhibit BRD4 and modulate the inflammatory response in Kupffer cells offers a promising avenue for future clinical intervention.
文摘With the wide application of thrombolytic drugs and the advancement of endovascular therapeutic techniques, the recanalization treatment of acute artery occlusion in ischemic stroke (IS) has made a leap forward, but ischemic brain tissues still face ischemia-reperfusion injury after recanalization. Nowadays, effective neurological protective agents still cannot completely resist the multiple damages of ischemia-reperfusion injury. As an iron-dependent mode of programmed cell death, ferroptosis occupies an important position in ischemia-reperfusion injury. Selenium plays a unique protective role in ischemia-reperfusion injury as an active site element in the center of glutathione peroxidase. Therefore, the study mainly aims to review the protective role of selenium in IS and the related mechanisms, as well as the effect of selenium on the risk factors of IS.
基金Supported by the National Natural Science Foundation of China(No.82071312).
文摘AIM:To explore the immune cell infiltration and molecular mechanisms of retinal ischemia-reperfusion injury(RIRI)to identify potential therapeutic targets.METHODS:In the bulk RNA-seq analysis,This study performed differential gene expression analysis,weighted gene co-expression network analysis,and protein-protein interaction network analysis to identify hub genes.QuanTIseq was used to determine the composition of infiltrating immune cells.Following the identification of hub genes,single-cell RNA-seq analysis was employed to pinpoint the specific immune cell types expressing these hub genes.Cell-cell communication analysis to explore signaling pathways and interactions between immune cells was further performed.Finally,the expression of these key immune regulators in vivo using quantitative real-time polymerase chain reaction(qRT-PCR)was validated.RESULTS:Bulk RNA-seq analysis identified Stat2,Irf7,Irgm1,Igtp,Parp9,Irgm2,Nlrc5,and Tap1 as hub genes,with strong correlations to immune cell infiltration.Single-cell RNA-seq analysis further revealed six immune cell clusters,showing Irf7 predominantly in microglia and Tap1 in dendritic cells(DCs).And cell-cell communication analysis showed that microglia and DCs play central roles in coordinating immune activity.qRT-PCR validated the upregulation of these genes.CONCLUSION:In the acute phase of RIRI,Irf7 and Tap1 may be the potential therapeutic targets to reduce inflammation and promote neurological function recovery.
文摘BACKGROUND Ferroptosis is a newly recognized form of regulated cell death characterized by iron-dependent accumulation of lipid reactive oxygen species.It has been extensively studied in various diseases,including cancer,Parkinson’s disease,and stroke.However,its precise role and underlying mechanisms in ischemia/reperfusion injury,particularly in the intestinal ischemia-reperfusion(IIR),remain unclear.In current work,we aimed to investigate the participation of histone lactylation during IIR progression.AIM To investigate the role of mitochondrial alanyl-tRNA synthetase 2(AARS2)in ferroptosis and its epigenetic regulation of acyl-CoA synthetase long-chain family member 4(ACSL4)through histone lactylation during IIR injury.METHODS We established a mouse model to mimic IIR and conducted AARS2 knockdown as treatment.The expression of AARS2 in intestinal tissues was measured by western blot.The integrity of intestinal tissues was detected by hematoxylin and eosin staining,serum fatty acid-binding protein,protein levels of ZO-1 and occluding.An in vitro hypoxia-reperfusion(H/R)cell model was established,and cell viability was measured by CCK-8.The in vitro and in vivo ferroptosis was determined by the accumulation of Fe2+and malondialdehyde(MDA).The epigenetic regulation of ACSL4 by AARS2 was detected by chromatin immunoprecipitation(ChIP)assay and luciferase reporter assay.RESULTS We observed a notable elevated AARS2 level in intestinal tissue of mice in IIR model group,which was reversed by shAARS2 treatment.Knockdown of AARS2 repressed alleviated intestinal barrier disruption and repressed the accumulation of ferroptosis biomarker Fe2+and MDA during IIR.The in vitro results showed that shAARS2 alleviated impaired cell viability caused by H/R,as well as repressed ferroptosis.Knockdown of AARS2 notably downregulated the RNA and protein expression of ACSL4.Mechanistically,knockdown of AARS2 downregulated the enrichment of H3K18 La modification on AARS2,as well as suppressed its promoter activity.Overexpression of AARS2 could abolish the protective effects of shACSL4 in vitro.CONCLUSION The elevation of AARS2 during IIR led to cell ferroptosis via epigenetically upregulating the expression of ACSL4.Our findings presented AARS2 as a promising therapeutic target for IIR.
基金Natural Science Foundation of Fujian Province,Grant/Award Number:2021J011262 and 2022J011095Youth Independent Innovation and Incubation Special Project,Grant/Award Number:2022QC07The project of 900th Hospital,Grant/Award Number:2021MS02 and 2023GK01。
文摘Background:The aim of the study was to explore a feasible method for alleviating limb ischemia-reperfusion injury(LI/RI)through the use of a high-concentration citrate solution(HC-A solution)for limb perfusion(LP).Methods:Eighteen pigs were divided into three groups:the Sham group,LI/RI group,and HCA group.The Sham group underwent exposure of the iliac artery and vein.The LI/RI group underwent tourniquet placement and clamping of the iliac artery and vein to simulate LI/RI.The HCA group received HC-A solution LP for 30 min through the left iliac artery below the level of blood flow occlusion based on the LI/RI group.Oxidative stress markers and inflammatory response markers were compared among the three groups.Results:Compared to the LI/RI group,the HCA group showed significantly lower levels of serum creatine kinase(CK),lactate dehydrogenase(LDH),malondialdehyde(MDA),tumor necrosis factor-α(TNF-α),aspartate aminotransferase(AST),and ala-nine aminotransferase(ALT),and significantly greater activities of serum superoxide dismutase(SOD)(p<0.05).There were no significant differences in serum interleukin-6(IL-6)or in muscle MDA,SOD,TNF-α,and IL-6 between the HCA group and the LI/RI group(p>0.05).Compared to the LI/RI group,MDA,TNF-α,and IL-6 levels in the liver were significantly lower in the HCA group(p<0.05),while SOD activities were not significantly different(p>0.05).Histopathological examination revealed reduced skeletal muscle and liver damage in the HCA group compared to the LI/RI group.Conclusions:HC-A solution LP can alleviate liver damage caused by LI/RI in pigs.
基金Supported by the National Natural Science Foundation of China,No.82200658.
文摘BACKGROUND Hepatic ischemia-reperfusion(I/R)injury related to liver transplantation and hepatic resection remains a challenge in clinical practice.Accumulating evidence indicates that mitochondrial dysfunction is a critical cause of I/R injury.The protein 4-nitrophenylphosphatase domain and non-neuronal SNAP25-like protein homolog 1(NIPSNAP1)is involved in the regulation of mitophagy and the recruitment of autophagy receptor proteins independent of PTEN induced putative kinase 1.AIM To clarify the protective mechanism of NIPSNAP1 against hepatic I/R,with a focus on mitophagy and mitochondrial dynamics,as well as the potential mechanism by which n6-methyladenosine(m6A)modification regulates NIPSNAP1.METHODS Mice were administered an adeno-associated virus in vivo and a hepatic I/R model was established via portal vein interruption followed by reperfusion to explore the effect of NIPSNAP1 on hepatic I/R.HepG2 cells were subjected to hypoxia/reoxygenation treatment in vitro.RESULTS We observed a significant downregulation of both NIPSNAP1 and insulin-like growth factor 2 mRNA-binding protein 2(IGF2BP2)expression in vivo and in vitro.NIPSNAP1 knockdown impaired mitophagy and disrupted mitochondrial dynamics;in contrast,NIPSNAP1 overexpression resulted in the opposite effects.Further studies revealed that IGF2BP2 functions as an m6A reader that targets and binds NIPSNAP1,thereby regulating its mRNA stability.CONCLUSION NIPSNAP1 prevents hepatic I/R injury by promoting mitophagy and maintaining mitochondrial homeostasis,serving as a novel target of the m6A reader IGF2BP2.Therefore,targeting the IGF2BP2/NIPSNAP1 axis may facilitate the development of better therapeutics for hepatic I/R.
基金Natural Science Foundation of Guangxi(Grant No.2021JJD140147)。
文摘PI3K/AKT/mTOR signaling pathway is a key pathway of myocardial ischemia-reperfusion injury(MIRI).The mechanism of action is mainly oxidative stress,inflammatory response,calcium overload,ferroptosis,autophagy,and apoptosis.MIRI belongs to the category of chest obstruction in traditional Chinese medicine,and its etiology and pathogenesis are mainly“Yang Wei Yin Xian.”Traditional Chinese medicine has the effect of multi-target and multi-component effect,and has played a significant role in the treatment of MIRI in recent years.At present,the monomers of traditional Chinese medicine mainly include saponins,flavonoids,alkaloids,terpenoids,and phenols,and the compounds mainly include Zhigancao Decoction,Zhenyuan Capsule,Jiawei Shenqibai Powder,Qili Qiangxin Capsule,Tongmai Yangxin Pill,Zhilong Huoxue Tongyu Capsule,Guizhi Tongluo Tablets,etc.This paper reviews the research on the improvement of MIRI by regulating PI3K/AKT/mTOR signaling pathway in recent years,and expounds the mechanism and advantages of traditional Chinese medicine in the treatment of MIRI.
基金supported the National Natural Science Foundation of China(81930113).
文摘Objective:To investigate the potential protective effect of Shexiang Tongxin dropping pills(STDP)on ischemia-reperfusion injury and its underlying mechanisms in improving endothelial cell function in coronary microvascular disease(CMVD).Methods:A rat model of myocardial ischemia-reperfusion injury with CMVD was established using ligation and reperfusion of the left anterior descending artery.The effect of STDP(21.6 mg/kg)on cardiac function was evaluated using echocardiography,hematoxylin-eosin staining,and Evans blue staining.The effects of STDP on the microvascular endothelial barrier were assessed based on nitric oxide production,endothelial nitric oxide synthase expression,structural variety of tight junctions(TJs),and the expression of zonula occludens-1(ZO-1),claudin-5,occludin,and vascular endothelial(VE)-cadherin proteins.The mechanisms of STDP(50 and 100 ng/mL)were evaluated by examining the expression of sphingosine 1-phosphate receptor 2(S1PR2),Ras Homolog family member A(RhoA),and Rho-associated coiled-coil-containing protein kinase(ROCK)proteins and the distribution of ZO-1,VE-cadherin,and Factin proteins in an oxygen and glucose deprivation/reoxygenation model.Results:The administration of STDP on CMVD rat model significantly improved cardiac and microvascular endothelial cell barrier functions(all P<.05).STDP enhanced the structural integrity of coronary microvascular positioning and distribution by clarifying and completing TJs and increasing the expression of ZO-1,occludin,claudin-5,and VE-cadherin in vivo(all P<.05).The S1PR2/RhoA/ROCK pathway was inhibited by STDP in vitro,leading to the regulation of endothelial cell TJs,adhesion junctions,and cytoskeletal morphology.Conclusion:STDP showed protective effects on cardiac impairment and microvascular endothelial barrier injury in CMVD model rats induced by myocardial ischemia-reperfusion injury through the modulation of the S1PR2/RhoA/ROCK pathway.
基金Natural Science Foundation of Anhui ProvinceGrant/Award Number:1908085MH241。
文摘Lung ischemia-reperfusion injury(LIRI),an acute lung injury syndrome triggered by lung transplantation or distal organ ischemia,has long been a difficult and hot issue in clinical research.In this study,we proposed a simple and less invasive reversible LIRI surgical protocol,achieved by improving the mouse left hilar entrapment model,which significantly improved the operability of the experiment and the reproducibility of the results.The protocol achieves precise control of the ischemic and reperfusion processes by visualizing transoral intubation,using reversible ligation of live knots to clamp the left hilar,and temporary closure of the thoracic cavity during ischemia.The reversible survival model we constructed not only provides a reliable tool to study the cellular and molecular mechanisms of LIRI but also can be used to assess the stage of injury regression,experimental pneumonia,and survival in mice.In addition,it simplifies the lung portal separation clamping operation for reversible clamping and provides an easy-to-learn visual tracheal intubation method that can be quickly mastered and replicated by beginners for consistent and reliable results.
基金Supported by Swedish Cancer society(Cancerfonden)the Swedish Cancer and Allergy fund(Cancer-och Allergifonden)
文摘AIM To study the effects of warm ischemia-reperfusion(I/R) injury on hepatic morphology at the ultrastructural level and to analyze the expression of the thioredoxin(TRX)and glutaredoxin(GRX) systems.METHODS Eleven patients undergoing liver resection were subjected to portal triad clamping(PTC). Liver biopsies were collected at three time points; first prior to PTC(baseline), 20 min after PTC(post-ischemia) and 20 min after reperfusion(post-reperfusion). Electron microscopy and morphometry were used to study and quantify ultrastructural changes, respectively. Additionally, gene expression analysis of TRX and GRX isoforms was performed by quantitative PCR. For further validation of redox protein status, immunogold staining was performed for the isoforms GRX1 and TRX1.RESULTS Post-ischemia, a significant loss of the liver sinusoidal endothelial cell(LSEC) lining was observed(P = 0.0003) accompanied by a decrease of hepatocyte microvilli in the space of Disse. Hepatocellular morphology was well preserved apart from the appearance of crystalline mitochondrial inclusions in 7 out of 11 patients. Postreperfusion biopsies had similar features as post-ischemia with the exception of signs of a reactivation of the LSECs. No changes in the expression of redox-regulatory genes could be observed at mR NA level of the isoforms of the TRX family but immunoelectron microscopy indicated a redistribution of TRX1 within the cell.CONCLUSION At the ultrastructural level, the major impact of hepatic warm I/R injury after PTC was borne by the LSECs with detachment and reactivation at ischemia and reperfusion, respectively. Hepatocytes morphology were well preserved. Crystalline inclusions in mitochondria were observed in the hepatocyte after ischemia.
基金supported by National Natural Science Foundation of China(No.81670334/No.81800262)Science and Technology Planning Project of Guangzhou(No.201903010005)+1 种基金Science and Technology Planning Project of Guangzhou(No.201906010089)Natural Science Foundation of Guangdong Province(No.2018A030313029)
文摘Background Myocardial ischemia/reperfusion(I/R)injury remains a great challenge in clinical therapy.Recent studies indicated that circularRNA(circRNA)play an important role in the regulation of atherosclerotic heart disease.However,as a new circRNA,the effect of circRNA regulating ischemia-reperfusion through microRNA-1(CRIRTM)on myocardial apoptosis during myocardial ischemia/reperfusion is still unknown.The purpose of this study was to investigate the effect of CRIRTM on I/R-induced apoptosis and the expression of microRNA-1(miR-1)and calmodulin(CaM)in cultured neonatal rat ventricular cardiomyocytes(NRVCs)in vitro and SD rat hypoxia-reoxygenation model in vivo.Methods NRVCs were subjected to hypoxia for 30 minutes and then oxygen was added for 12 hours to establish the model of cell ischemia reperfusion.The SD rats were subjected to the surgery of ligating the anterior descending branch of the left coronary artery for 4 hours and then reperfused for 48 hours to establish myocardial ischemia animal model.The expression of CRIRTM,miIR-1 and CaM was detected by quantitative real-time polymerase chain reaction(qRT-PCR).TUNEL was used to analyze the apoptosis level of cells and animal myocardium after small interfering RNA(siRNA)down-regulation of CRIRTM.The expression of CRIRTM in the blood of patients with myocardial infarction was detected,and the downstream of CRIRTM was analyzed by Target Scan.Results The specific upregulation of CRIRTM was detected in the plasma of patients with ST-segment elevation myocardial infarction after reperfusion,and multiple binding sites between CRIRTM and miR-1 were found by Target Scan analysis.The expression of CRIRTM and CaM was up-regulated and the expression of miR-1 was down-regulated in cells and rats I/R model.Both in vivo and in vitro,cardiomyocyte apoptosis induced by myocardial ischemia-reperfusion will increase after down-regulation of CRIRTM.It may be due to the decreased inhibition of miR-1 by down-regulated CRIRTM,which increases the degradation of CaM by miR-1 and aggravates cardiomyocyte apoptosis.Conclusions Our data demonstrated that CRIRTM alleviates myocardial ischemia-reperfusion injury via suppressing miR-1 leading to enhanced CaM levels.[S Chin J Cardiol 2021;22(1):30-37]
