BACKGROUND： Vascular endothelial cell apoptosis participates in cerebral ischemia/reperfusion injury, and the method of Qi-supplementation and blood-activation has remarkable neuroprotective effects against cerebral ...BACKGROUND： Vascular endothelial cell apoptosis participates in cerebral ischemia/reperfusion injury, and the method of Qi-supplementation and blood-activation has remarkable neuroprotective effects against cerebral ischemia/reperfusion injury. OBJECTIVE： To explore the molecular mechanism that Dingnaofang （Chinese herbs for supplementing Qi and activating blood circulation） inhibits vascular endothelial cell apoptosis induced by cerebral ischemia/reperfusion injury. DESIGN, TIME AND SETFING： A randomized controlled animal experiment was performed at the Department of Central Laboratory, Third Xiangya Hospital, Central South University, China, between October 2007 and December 2008. MATERIALS： Dingnaofang consisted of Huangqi （Milkvetch Root; Radix Astragah）, Chuanxiong （Szechwan Lovage Rhizome; Rhizoma Chuanxiong）, Yinxingye （ginkgo leaf; Fofium Ginkgo）, Dilong （earthworm; Pheretima）, Danggui （Chinese Angelica; Radix Angelicae Sinensis）, Tianqi （Radix Notoginseng）, and Gancao （Radix Glycyrrhizae; Radix Glycytthizae）, with a proportion of 5：2：2： 1： 1： 1： 1. METHODS： A total of 130 Sprague Dawley rats were randomly divided into sham-surgery （n = 10）, cerebral ischemia/reperfusion （n = 40）, cerebral ischemia pretreatment （n = 40） and Dingnaofang pretreatment groups （n = 40）. Middle cerebral artery occlusion was used to induce cerebral ischemic injury. The bilateral common carotid artery in the cerebral ischemia pretreatment group was blocked for 10 minutes on days 7, 3 and 1 prior to ischemia/reperfusion injury, while rats in Dingnaofang pretreatment group were intragastrically administrated with 4 g Dingnaofang 1 week prior to cerebral ischemia once per day, for 7 successive days. MAIN OUTCOME MEASURES： Apoptosis ratios in vascular endothelial cells were measured using Hoechst 33258 staining and flow cytometry; apoptosis was detected by monitoring DNA gradient bands and the activation of caspase-3, 8, 9 and Bid using Western blot. RESULTS： Following cerebral ischemiaJreperfusion injury, the number of apoptotic vascular endothelial cells in the middle cerebral artery significantly increased （P 〈 0.01）; however, cerebral ischemia pretreatment and Dingnaofang pretreatment groups significantly reduced apoptosis induced by cerebral ischemia/reperfusion injury （P 〈 0.01）. In particular, DNA gradient bands were not observed following Dingnaofang pretreatment. At 24 hours after cerebral ischemia/reperfusion injury, cleaved fragments of caspase-3, 8 and 9 were detected at 11 kD （P 11）, 20 kD （P 20） and 10 kD （P 10）, respectively, following Western blot. Bid was also cleaved into its truncated form （tBid; 15 kD）. Gray scale analysis indicated that P 11, P 20, P 10 and tBid band values in the Dingnaofang pretreatment group were significantly less than in the cerebral ischemia/reperfusion and cerebral ischemia pretreatment groups （P 〈 0.01 or P 〈 0.05）. CONCLUSION： Dingnaofang inhibits vascular endothelial cell apoptosis induced by cerebral ischemia/reperfusion injury via inhibition of apoptotic signal transduction pathways.展开更多
Ischemia–reperfusion injury is a common pathophysiological mechanism in retinal degeneration.PANoptosis is a newly defined integral form of regulated cell death that combines the key features of pyroptosis,apoptosis,...Ischemia–reperfusion injury is a common pathophysiological mechanism in retinal degeneration.PANoptosis is a newly defined integral form of regulated cell death that combines the key features of pyroptosis,apoptosis,and necroptosis.Oligomerization of mitochondrial voltage-dependent anion channel 1 is an important pathological event in regulating cell death in retinal ischemia–reperfusion injury.However,its role in PANoptosis remains largely unknown.In this study,we demonstrated that voltage-dependent anion channel 1 oligomerization-mediated mitochondrial dysfunction was associated with PANoptosis in retinal ischemia–reperfusion injury.Inhibition of voltage-dependent anion channel 1 oligomerization suppressed mitochondrial dysfunction and PANoptosis in retinal cells subjected to ischemia–reperfusion injury.Mechanistically,mitochondria-derived reactive oxygen species played a central role in the voltagedependent anion channel 1-mediated regulation of PANoptosis by promoting PANoptosome assembly.Moreover,inhibiting voltage-dependent anion channel 1 oligomerization protected against PANoptosis in the retinas of rats subjected to ischemia–reperfusion injury.Overall,our findings reveal the critical role of voltage-dependent anion channel 1 oligomerization in regulating PANoptosis in retinal ischemia–reperfusion injury,highlighting voltage-dependent anion channel 1 as a promising therapeutic target.展开更多
Several studies have shown that activation of unfolded protein response and endoplasmic reticulum(ER)stress plays a crucial role in severe cerebral ischemia/reperfusion injury.Autophagy occurs within hours after cereb...Several studies have shown that activation of unfolded protein response and endoplasmic reticulum(ER)stress plays a crucial role in severe cerebral ischemia/reperfusion injury.Autophagy occurs within hours after cerebral ischemia,but the relationship between ER stress and autophagy remains unclear.In this study,we established experimental models using oxygen-glucose deprivation/reoxygenation in PC12 cells and primary neurons to simulate cerebral ischemia/reperfusion injury.We found that prolongation of oxygen-glucose deprivation activated the ER stress pathway protein kinase-like endoplasmic reticulum kinase(PERK)/eukaryotic translation initiation factor 2 subunit alpha(e IF2α)-activating transcription factor 4(ATF4)-C/EBP homologous protein(CHOP),increased neuronal apoptosis,and induced autophagy.Furthermore,inhibition of ER stress using inhibitors or by si RNA knockdown of the PERK gene significantly attenuated excessive autophagy and neuronal apoptosis,indicating an interaction between autophagy and ER stress and suggesting PERK as an essential target for regulating autophagy.Blocking autophagy with chloroquine exacerbated ER stress-induced apoptosis,indicating that normal levels of autophagy play a protective role in neuronal injury following cerebral ischemia/reperfusion injury.Findings from this study indicate that cerebral ischemia/reperfusion injury can trigger neuronal ER stress and promote autophagy,and suggest that PERK is a possible target for inhibiting excessive autophagy in cerebral ischemia/reperfusion injury.展开更多
Nitric oxide(NO)is a gaseous molecule produced by 3 different NO synthase(NOS)isoforms:Neural/brain NOS(nNOS/bNOS,type 1),endothelial NOS(eNOS,type 3)and inducible NOS(type 2).Type 1 and 3 NOS are constitutively expre...Nitric oxide(NO)is a gaseous molecule produced by 3 different NO synthase(NOS)isoforms:Neural/brain NOS(nNOS/bNOS,type 1),endothelial NOS(eNOS,type 3)and inducible NOS(type 2).Type 1 and 3 NOS are constitutively expressed.NO can serve different purposes:As a vasoactive molecule,as a neurotransmitter or as an immunomodulator.It plays a key role in cerebral ischemia/reperfusion injury(CIRI).Hypoxic episodes simulate the production of oxygen free radicals,leading to mitochondrial and phospholipid damage.Upon reperfusion,increased levels of oxygen trigger oxide synthases;whose products are associated with neuronal damage by promoting lipid peroxidation,nitrosylation and excitotoxicity.Molecular pathways in CIRI can be altered by NOS.Neuroprotective effects are observed with eNOS activity.While nNOS interplay is prone to endothelial inflammation,oxidative stress and apoptosis.Therefore,nNOS appears to be detrimental.The interaction between NO and other free radicals develops peroxynitrite;which is a cytotoxic agent.It plays a main role in the likelihood of hemorrhagic events by tissue plasminogen activator(t-PA).Peroxynitrite scavengers are currently being studied as potential targets to prevent hemorrhagic transformation in CIRI.展开更多
Cerebral ischemia/reperfusion(I/R)injury is an important pathophysiological condition of ischemic stroke that involves a variety of physiological and pathological cell death pathways,including autophagy,apoptosis,necr...Cerebral ischemia/reperfusion(I/R)injury is an important pathophysiological condition of ischemic stroke that involves a variety of physiological and pathological cell death pathways,including autophagy,apoptosis,necroptosis,and phagoptosis,among which autophagy is the most studied.We have reviewed studies published in the past 5 years regarding the association between autophagy and cerebral I/R injury.To the best of our knowledge,this is the first review article summarizing potential candidates targeting autophagic pathways in the treatment of I/R injury post ischemic stroke.The findings of this review may help to better understand the pathogenesis and mechanisms of I/R events and bridge the gap between basic and translational research that may lead to the development of novel therapeutic approaches for I/R injury.展开更多
Background:Ex vivo lung perfusion(EVLP)has emerged as a critical technique for lung preservation and evaluation prior to transplantation.While conventional rat EVLP systems utilize closed-loop dual cannulation of pulm...Background:Ex vivo lung perfusion(EVLP)has emerged as a critical technique for lung preservation and evaluation prior to transplantation.While conventional rat EVLP systems utilize closed-loop dual cannulation of pulmonary artery(PA)and vein,the effect of the simplified model using single PA cannulation with passive venous drainage is unknown.Methods:We developed two EVLP models in rats:a semi-closed circuit with PA-only cannulation and left atrial incision for passive venous drainage(SC-EVLP),and a closed circuit employing both arterial and venous cannulation(C-EVLP).Donor lungs were perfused for a defined duration and subsequently orthotopically transplanted.We evaluated pulmonary function parameters,histopathological injury scores,inflammatory cytokine levels,and apoptotic marker expression at the end of perfusion and posttransplantation.Results:Compared to the conventional EVLP,the SC-EVLP group exhibited significantly lower PA pressure and improved dynamic lung compliance throughout perfusion.Although the levels of tumor necrosis factor-αin the perfusate were higher in the SC-EVLP group,other cytokine levels in the perfusate and bronchoalveolar lavage fluid exhibited no significant differences.Pulmonary edema was reduced in the SC-EVLP group,as indicated by a lower lung wet-to-dry ratio.After transplantation,lungs from the SC-EVLP group exhibited lower histological injury scores,reduced apoptosis,and decreased serum cytokine levels,suggesting attenuated inflammation and tissue damage.Conclusions:In a rat model,single PA cannulation with passive venous drainage reduced pulmonary edema during EVLP and reduced lung injury and systemic inflammation after transplantation.展开更多
Objective Cerebral venous outflow disorders(CVOD)can impair cerebral perfusion and produce diverse,often debilitating symptoms,substantially reducing quality of life.Intermittent hypoxiahyperoxia training(IHHT)has dem...Objective Cerebral venous outflow disorders(CVOD)can impair cerebral perfusion and produce diverse,often debilitating symptoms,substantially reducing quality of life.Intermittent hypoxiahyperoxia training(IHHT)has demonstrated therapeutic potential across various pathologies and may represent a promising non-pharmacological approach for CVOD management.Methods Patients with imaging-confirmed CVOD underwent 14 IHHT sessions,each comprising four cycles of 10-minute hypoxia(11%O_(2))stimulation and 20-minute hyperoxia(38%O_(2)).Physiological parameters and adverse events were monitored throughout the intervention.Clinical scales,24-hour ambulatory blood pressure,blood tests,jugular ultrasound,and perfusion imaging were assessed preand post-intervention.Results No participants experienced intolerable discomfort or severe adverse events;vital signs remained within normal ranges.No significant changes were observed in 24-hour blood pressure,blood cell counts,lipid profiles,or other blood markers.Notably,60%of patients(n=12)reported overall symptom improvement on the Patient Global Impression of Change scale.Headache severity,as measured by the visual analogue scale,significantly decreased(6.33±1.22 vs.4.89±2.03,P=0.016).In patients with internal jugular vein(IJV)stenosis,significant improvements were observed in regional cerebral blood flow(including the insula,occipital lobe,internal capsule,and lenticula)and left J3-segment IJV flow volume(107.27[47.50,160.00]vs.140.83[55.00,210.00]mL/min,P=0.011).Conclusion The current IHHT protocol is safe and well-tolerated in patients with CVOD.IHHT may alleviate CVOD-related symptoms by improving oxygen saturation,cerebral perfusion,and venous outflow pattern,supporting its potential as a non-invasive therapeutic strategy.展开更多
Objective:Acute kidney injury(AKI)is a clinical syndrome characterized by sudden deterioration of renal function,with ischemia reperfusion injury(IRI)being the most common cause.Long noncoding RNA(lncRNA)regulate cell...Objective:Acute kidney injury(AKI)is a clinical syndrome characterized by sudden deterioration of renal function,with ischemia reperfusion injury(IRI)being the most common cause.Long noncoding RNA(lncRNA)regulate cell fate through interactions with microRNA(miRNA)and messenger RNAs(mRNA),but the mechanisms and regulatory networks underlying lncRNA AK154753(AK154753)in IRI-induced AKI remain unclear.This study aims to investigate the role of AK154753 in acute renal IRI and to elucidate the molecular mechanism of the AK154753 via miR-345-3p/Bcl-2 homologous antagonist/killer(Bak)and miR-708-5p/Bcl-2 interacting mediator of cell death(Bim)axis.Methods:A bilateral renal artery ischemia model was established in mice(30 minutes ischemia followed by 24 hours and 48 hours reperfusion).Kidney tissues were analyzed using microarray-based transcriptomic sequencing to identify differentially expressed lncRNAs,miRNAs,and mRNAs.RNA levels of AK154753,miR-345-3p,miR-708-5p,Bak,and Bim were validated using real-time reverse transcription PCR(real-time RTPCR).Oxygen and glucose deprivation/reperfusion(OGD/R)models were constructed in mouse proximal renal tubular epithelial BUMPT cells to simulate in vitro IRI conditions.Adeno-associated virus(AAV)-mediated shRNA was used to silence AK154753 in vivo.Apoptosis was assessed using TUNEL staining and flow cytometry.Protein levels of Bak,Bim,and cleaved-caspase-3 were measured using Western blotting.Fluorescence in situ hybridization(FISH)was used to determine intracellular localization of AK154753.Binding relationships between AK154753 and miR-345-3p/Bak and miR-708-5p/Bim were verified using dual-luciferase reporter assays.MiRNA mimics and inhibitors were used to evaluate regulatory-network integrity.Results:IRI significantly elevated serum blood urea nitrogen(BUN)and serum creatinine(Scr),accompanied by tubular-structure damage and increased cell apoptosis(all P<0.05).Transcriptome profiling and real-time RT-PCR validation demonstrated that lncRNA AK154753,along with the pro-apoptotic proteins Bak and Bim,was significantly upregulated after IRI,whereas miR-345-3p and miR-708-5p were markedly downregulated(P<0.01).In vitro,OGD/R treatment significantly induced AK154753 expression in renal tubular epithelial cells and suppressed the expression of miR-345-3p and miR-708-5p,while markedly increasing the protein levels of Bak,Bim,and cleaved-caspase 3,resulting in a significant increase in apoptosis(all P<0.01).Silencing AK154753 significantly attenuated OGD/R-induced apoptosis,reduced the expression of Bak,Bim,and cleaved caspase 3,and decreased cell apoptosis(all P<0.01),while significantly upregulating miR-345-3p and miR-708-5p expression(P<0.01).In vivo,adeno-associated virus(AAV)-mediated knockdown of AK154753 significantly improved renal function in IRI mice,alleviated tubular injury,and suppressed renal tissue apoptosis,as evidenced by reduced BUN and Scr levels,improved histopathological injury scores,and decreased expression of Bak,Bim,and cleaved caspase-3(all P<0.01),accompanied by significant upregulation of miR-345-3p and miR-708-5p(all P<0.01).Luciferase reporter assays further confirmed that miR-345-3p directly binds to the 3'-untranslated region(3'-UTR)of AK154753 and Bak,whereas miR-708-5p directly binds to the 3'-UTRs of AK154753 and Bim.Inhibition of miR-345-3p or miR-708-5p abolished the anti-apoptotic effects induced by AK154753 silencing and restored Bak and Bim expression levels(all P<0.01).Conclusion:AK154753 is upregulated in acute renal IRI and promotes apoptosis by suppressing miR-345-3p and miR-708-5p,thereby upregulating Bak and Bim,and participates in the initiation and progression of acute renal IRI.展开更多
Introduction: Renal ischemia-reperfusion (IR) is responsible for injuries such as destruction or dysfunction of tubular epithelial cells with inflammatory reaction and oxidative stress. Several therapeutic methods hav...Introduction: Renal ischemia-reperfusion (IR) is responsible for injuries such as destruction or dysfunction of tubular epithelial cells with inflammatory reaction and oxidative stress. Several therapeutic methods have been tested to alleviate ischemia-perfusion injury, ranging from using anti-inflammatory drugs, antioxidants, and plants from traditional pharmacopeia to administering RNA interference. However, there is currently no effective therapeutic option available for the treatment of renal IR injury, other than supportive therapies such as renal replacement therapy or hydration. Objective: This present study aimed to evaluate the effect of Guiera senegalensis on renal ischemia reperfusion, a recognized plant for its antioxidant and anti-inflammatory properties. Materials and Methods: Twenty-four (24) adult male Wistar rats were divided into four following groups: SLAM (subjected to a median laparotomy with simulated ischemia);GUIERRA (animals that received 250 mg/kg of guierra senegalensis orally, once a day, for 5 days, with simulated renal ischemia);IR (animals that underwent laparotomy followed by clamping of bilateral renal pedicles for 45 min and followed by reperfusion);GUIERRA + IR (animals given GUIERRA at the dosage of 250 mg/kg per day, for 5 days and then subjected to renal ischemia-reperfusion). Data analysis was performed by ANOVA, and a significance level of p Results: Compared with the I/R group, rats in the GUIERRA + IR group showed reduced histopathological damage scores (p Conclusion: The results of this preliminary work suggest that Guiera senegalensis decreases the degree of tissue damage in renal ischemia-reperfusion cases. This plant seems to be a promising therapeutic;further studies could help to precise the targets of its compounds on ischemia-reperfusion pathways.展开更多
Vascular endothelial growth factor and its mimic peptide KLTWQELYQLKYKGI(QK)are widely used as the most potent angiogenic factors for the treatment of multiple ischemic diseases.However,conventional topical drug deliv...Vascular endothelial growth factor and its mimic peptide KLTWQELYQLKYKGI(QK)are widely used as the most potent angiogenic factors for the treatment of multiple ischemic diseases.However,conventional topical drug delivery often results in a burst release of the drug,leading to transient retention(inefficacy)and undesirable diffusion(toxicity)in vivo.Therefore,a drug delivery system that responds to changes in the microenvironment of tissue regeneration and controls vascular endothelial growth factor release is crucial to improve the treatment of ischemic stroke.Matrix metalloproteinase-2(MMP-2)is gradually upregulated after cerebral ischemia.Herein,vascular endothelial growth factor mimic peptide QK was self-assembled with MMP-2-cleaved peptide PLGLAG(TIMP)and customizable peptide amphiphilic(PA)molecules to construct nanofiber hydrogel PA-TIMP-QK.PA-TIMP-QK was found to control the delivery of QK by MMP-2 upregulation after cerebral ischemia/reperfusion and had a similar biological activity with vascular endothelial growth factor in vitro.The results indicated that PA-TIMP-QK promoted neuronal survival,restored local blood circulation,reduced blood-brain barrier permeability,and restored motor function.These findings suggest that the self-assembling nanofiber hydrogel PA-TIMP-QK may provide an intelligent drug delivery system that responds to the microenvironment and promotes regeneration and repair after cerebral ischemia/reperfusion injury.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
Objective:To evaluate the cardioprotective effects of sinomenine using the ischemia/reperfusion(I/R)rat model.Methods:Wistar rats were randomly divided into 6 groups:group Ⅰ with reperfusion,group Ⅱ perfused with si...Objective:To evaluate the cardioprotective effects of sinomenine using the ischemia/reperfusion(I/R)rat model.Methods:Wistar rats were randomly divided into 6 groups:group Ⅰ with reperfusion,group Ⅱ perfused with sinomenine,group Ⅲ perfused with 5-hydroxydecanoate(5-HD),group Ⅳ perfused with 5-HD+sinomenine,group Ⅴ perfused with L-nitro arginine methyl ester(L-NAME),group Ⅵ perfused with L-NAME+sinomenine.Myocardial ischemia was induced by interrupting the aortic blood supply for 30 min,followed by reperfusion(55 min).Cardiac,hepatic,antioxidant,and inflammatory parameters were assessed.Additionally,endothelin,tissue factor,platelet-activating factor,plasminogen activator inhibitor,plasma fibrinogen,and thromboxane B2 were also analyzed.Results:Administration of 5-HD or L-NAME,used as the selective antagonist of mitoKATP and NO system,respectively,resulted in significantly increased levels of premature ventricular complexes,lactate dehydrogenase,ventricular fibrillation,ventricular tachycardia,and arrhythmia intensity(P<0.05).In contrast,sinomenine significantly reduced the level of troponin Ⅰ,lactate dehydrogenase,creatine kinase,and creatine kinase MB compared to the 5-HD group and the L-NAME group(P<0.05).Additionally,sinomenine significantly reduced malondialdehyde level and enhanced the levels of superoxide dismutase,glutathione peroxidase,catalase,and glutathione/glutathione disulfide ratio(P<0.05).It also significantly suppressed the levels of endothelin-1,platelet-activating factor,tissue factor,plasminogen activator inhibitor 1,thromboxane B2,and plasma fibrinogen(P<0.05).Conclusions:These results suggest that sinomenine exhibits significant cardioprotection effects against I/R-induced cardiac injury in rats.展开更多
Background:Astrocyte endfeet(AEF)serves as a key element of the blood-brain barrier and is important for the survival and maintenance of neuronal function.However,the immunohistochemical and ultrastructural changes of...Background:Astrocyte endfeet(AEF)serves as a key element of the blood-brain barrier and is important for the survival and maintenance of neuronal function.However,the immunohistochemical and ultrastructural changes of AEF in the CA1 and CA3 areas of the hippocampus over time following cerebral ischemia-reperfusion(IR)injury have not been well elucidated.Objectives:We investigated chronological changes in AEF in the gerbil hippocampal CA1 area from 3 h to 10 days following transient forebrain ischemia(TFI),and examined their association with neuronal death and tissue repair following IR injury.Changes in the CA3 area were also examined at 10 days post-TFI for comparative purposes.Methods:Neuronal death was confirmed using histochemistry,immunohistochemistry,and histofluorescence.Changes in AEF were examined by double immunofluorescence with glial fibrillary acidic protein(GFAP)and glucose transporter 1(GLUT1),and by transmission electron microscopy(TEM)for ultrastructural changes.Results:Significant TFI-induced neuronal death occurred in the CA1 area on day 5 following IR injury and persisted until 10 days after TFI,while no neuronal death(or loss)was found in the CA3 area after TFI.Looking at TFI-induced changes in AEF,at 3 and 6 h after TFI,GFAP-immunoreactive(+)AEF in the CA1 area appeared swollen and harbored enlarged,dark mitochondria,and the swelling was reduced by 1-day post-TFI.On 2 and 5 days following TFI,GFAP+AEF were markedly enlarged and fragmented,containing shrunken mitochondria,vacuolations,and sparse organelles.Ten days post-TFI,the ends of GFAP+astrocytic processes extended to microvessels,appeared edematous,and were filled with cellular debris.In the CA3 area,AEF was slightly dilated at 10 days after TFI.These findings indicate that damage to or disruption of AEF in the CA1 area occurs in the early phase after 5-min TFI but is rarely observed in the CA3 area.Conclusion:Taken together,damage to or disruption of AEF following ischemic insults may be strongly linked to neuronal death/loss.展开更多
BACKGROUND Myocardial ischemia/reperfusion(I/R)injury,which is associated with high morbidity and mortality,is a main cause of unexpected myocardial injury after acute myocardial infarction.However,the underlying mech...BACKGROUND Myocardial ischemia/reperfusion(I/R)injury,which is associated with high morbidity and mortality,is a main cause of unexpected myocardial injury after acute myocardial infarction.However,the underlying mechanism remains unclear.Circular RNAs(circRNAs),which are formed from protein-coding genes,can sequester microRNAs or proteins,modulate transcription and interfere with splicing.Authoritative studies suggest that circRNAs may play an important role in myocardial I/R injury.AIM To explore the role and mechanism of circRNAs in myocardial I/R injury.METHODS We constructed a myocardial I/R injury model using ligation of the left anterior descending coronary artery,and evaluated the success of the validated model using triphenyltetrazolium chloride and hematoxylin-eosin staining.Then,left ventricular samples from different groups were selected for mRNA-sequence,and differential gene screening was performed on the obtained results.The differentially obtained mRNAs were divided into up-regulated and down-regulated according to their expression levels,and Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)functional enrichment analysis were performed,respectively.Then,the obtained circRNA and microRNA(miRNA)were paired for analysis,and the binding sites of miRNA and mRNA were virtual screened.Finally,the obtained circRNA,miRNA and mRNA were constructed by ceRNA mutual most useful network.RESULTS We used an RNA sequencing array to investigate the expression signatures of circRNAs in myocardial I/R injury using three samples from the I/R group and three samples from the sham group.A total of 142 upregulated and 121 downregulated circRNAs were found to be differentially expressed(fold change≥2,P<0.05).GO and KEGG functional analyses of these circRNAs were performed.GO analysis revealed that these circRNAs were involved mainly in cellular and intracellular processes.KEGG analysis demonstrated that 6 of the top 20 pathways were correlated with cell apoptosis.Furthermore,a circRNA-miRNA coexpression network and ceRNA network based on these genes were constructed,revealing that mmu-circ-0001452,mmu-circ-0001637,and mmu-circ-0000870 might be key regulators of myocardial I/R injury.CONCLUSION This research provides new insights into the mechanism of myocardial I/R,which mmu-circ-0001452,mmu-circ-0001637,and mmu-circ-0000870 are expected to be new therapeutic targets for myocardial I/R injury.展开更多
Objectives:Skeletal muscle ischemia/reperfusion injury(IRI)occurs as a result of a marked reduction in arterial perfusion to a limb and can lead to tissue death and threaten limb viability.This work assessed the effec...Objectives:Skeletal muscle ischemia/reperfusion injury(IRI)occurs as a result of a marked reduction in arterial perfusion to a limb and can lead to tissue death and threaten limb viability.This work assessed the effects of 20-hydroxyecdysone(20E)on hindlimb skeletal tissue following tourniquet-induced ischemia/reperfusion injury.Methods:Animals were divided into 4 groups—control group(Control),Control+20E(C+20E),mice with IRI(IRI),and mice with IRI+20E(IRI+20E).IRI was modeled by applying a tourniquet to the hind limb for 2 h with reperfusion for 1 h.5 mg/kg of 20E was administered intraperitoneally for 14 days.Afterward,the physical activity of mice,the histological structure of the quadriceps femoris,the expression of genes encoding proteins induced by hypoxia and involved in tissue adaptation to ischemia,and the functional parameters of skeletal muscle mitochondria were assessed.Results:It was shown that IRI of the limbs leads to functional disorders,depression of muscle function,accumulation of malondialdehyde(MDA)in mitochondria,and a decrease in their Ca2+buffering capacity,as well as an increase in the expression of HIF-1α,VGEF-A,PGC1αand PDGF-BB genes associated with adaptation to ischemia.20E reduced the intensity of degenerative processes in skeletal muscles,which was expressed in a decrease in the number of centrally nucleated fibers.Analysis of gene expression levels indicated a high degree of adaptation of animals to IRI.20E reduced the level of MDA in mitochondria,but did not affect the rate of respiration and calcium retention capacity of organelles both in normal conditions and during IRI.Conclusion:20E partially alleviates the skeletal muscle damage caused by IRI and can be used as part of combination therapy.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
Hepatic ischemia/reperfusion injury(IRI)remains a critical complication contributing to graft dysfunction following liver surgery.As part of an ongoing search for hepatoprotective natural products,five previously unre...Hepatic ischemia/reperfusion injury(IRI)remains a critical complication contributing to graft dysfunction following liver surgery.As part of an ongoing search for hepatoprotective natural products,five previously unreported homoadamantane-type polycyclic polyprenylated acylphloroglucinols(PPAPs),named hyperhomanoons A-E(1-5),and one known analog,hypersampsone O(6),were isolated from Hypericum patulum.Among these,compound 6 demonstrated potent protective effects against CoCl_(2)-induced hypoxic injury in hepatocytes.Furthermore,in a murine model of hepatic IRI induced by vascular occlusion,pretreatment with 6 markedly alleviated liver damage and reduced hepatocyte apoptosis.This study is the first to identify PPAPs as promising scaffolds for the development of therapeutic agents targeting hepatic IRI,underscoring their potential as lead compounds in drug discovery efforts for ischemic liver diseases.展开更多
Increasing evidence of the significant clinical value of protection against ischemia/reperfusion injury has contributed to the realization of the independent importance of this approach in improving prognosis and redu...Increasing evidence of the significant clinical value of protection against ischemia/reperfusion injury has contributed to the realization of the independent importance of this approach in improving prognosis and reducing cardiovascular mortality.Extracellular vesicles(EVs)derived by adipose mesenchymal stem cells may mediate the paracrine effects of stem cells and provide regenerative and anti-inflammatory properties,which are enhanced byγ-aminobutyric acid.The protective effects on cardiac myocytes may result from the EV embarked by miR-21-5p,which is a target for thioredoxin-interacting protein,regulating the formation of thioredoxin-interacting protein-thioredoxin complexes and subsequently enhancing the antioxidant activity of thioredoxin.It has been found thatγ-aminobutyric acid governs myocardial bioenergetics through suppressing inflammation and supporting mitochondrial structure.Finally,stem cell-based cell-free therapy based on adipose-derived stem cell EVs is considered a promising approach to individualized management of ischemia-induced cardiomyopathy.展开更多
Objective MicroRNA-1(miR-1)aggravates myocardial ischemia–reperfusion(I/R)injury,whereas insulin-like growth factor-1(IGF-1)maintains cardiomyocyte homeostasis.In this study,the aim is to investigate whether miR-1 ca...Objective MicroRNA-1(miR-1)aggravates myocardial ischemia–reperfusion(I/R)injury,whereas insulin-like growth factor-1(IGF-1)maintains cardiomyocyte homeostasis.In this study,the aim is to investigate whether miR-1 can exacerbate I/R injury through the regulation of IGF-1.Methods The infarct area,lactate dehydrogenase,miR-1 level,and apoptosis level were examined in the Langendorff isolated rat I/R model.The hypoxia–reoxygenation model of rat cardiacmyocytes and H9c2 cells were developed to determine the levels of miR-1,IGF-1 mRNA,and IGF-1 protein.Furthermore,the dual-luciferase assay was used to verify the relationship between miR-1 and IGF-1.Results Overexpression of miR-1 increased the level of apoptosis and decreased the IGF-1 expression.However,inhibition of miR-1 expression decreased the level of apoptosis,alleviated the degree of injury,and increased the IGF-1 expression.Overexpression of IGF-1 also reduced the degree of cellular damage and level of apoptosis caused by the overexpression of miR-1.When IGF-1 was knocked down,myocardial cells displayed more severe damage and a higher apoptosis level,even with decreased levels of miR-1.Conclusion miR-1 promotes apoptosis and aggravates I/R injury by downregulating IGF-1.展开更多
BACKGROUND Hepatic ischemia reperfusion(HIR)injury is a major complication affecting various major liver surgeries,including liver transplantation.Aprepitant(APRE),a neurokinin-1 receptor antagonist,is commonly used a...BACKGROUND Hepatic ischemia reperfusion(HIR)injury is a major complication affecting various major liver surgeries,including liver transplantation.Aprepitant(APRE),a neurokinin-1 receptor antagonist,is commonly used as an antiemetic to prevent chemotherapy-induced nausea and vomiting.AIM To assess the potential protective effect of APRE against HIR-induced liver injury via targeting the nucleotide-binding oligomerization domain-,leucine-rich repeat-,and pyrin domain-containing receptor 3/interleukin(IL)-1beta signaling pathway.METHODS Six groups of adult male Wistar albino rats were divided as follows:Sham group,Sham/APRE10 group(APRE 10 mg/kg),HIR group,HIR/APRE5 group(APRE 5 mg/kg),HIR/APRE10 group(APRE 10 mg/kg),and HIR/APRE20 group(APRE 20 mg/kg).Serum alanine transaminase,aspartate transaminase,liver malondialdehyde,total antioxidant capacity levels,as well as IL-6,sirtuin 1(Sirt1),caspase-3,cleaved caspase-3,and tumor necrosis factor alpha biomarkers,were evaluated.Hepatic specimens were examined histopathologically and immunohistochemically for nuclear factor erythroid-2-related factor 2(Nrf2)immunoexpression.RESULTS HIR resulted in hepatic damage,as evidenced by histopathological changes and a significant increase in serum alanine transaminase,aspartate transaminase,hepatic malondialdehyde,caspase-3,and tumor necrosis factor alpha levels.Additionally,there were significant increases in hepatic total antioxidant capacity and reductions in IL-6 and cleaved caspase-3 protein levels,as demonstrated by Western blot analysis,along with enhanced immunoexpression of Sirt1 and Nrf2.APRE has significantly reduced various parameters of oxidative stress,inflammation,and apoptosis,and a significant increase in liver Nrf2 immunoexpression,leading to a significant improvement in the histopathological changes.CONCLUSION In conclusion,targeting the Sirt1/Nrf2 signaling pathway,as demonstrated by APRE in our model,could present a promising therapeutic target to protect against HIR-induced liver injury during major liver surgeries.展开更多
基金the National Natural Science Foundation of China,No. 30672035Scientific and Technological Foundation of Traditional Chinese Medicine of Public Health Bureau of Hunan Province,No. 620230
文摘BACKGROUND： Vascular endothelial cell apoptosis participates in cerebral ischemia/reperfusion injury, and the method of Qi-supplementation and blood-activation has remarkable neuroprotective effects against cerebral ischemia/reperfusion injury. OBJECTIVE： To explore the molecular mechanism that Dingnaofang （Chinese herbs for supplementing Qi and activating blood circulation） inhibits vascular endothelial cell apoptosis induced by cerebral ischemia/reperfusion injury. DESIGN, TIME AND SETFING： A randomized controlled animal experiment was performed at the Department of Central Laboratory, Third Xiangya Hospital, Central South University, China, between October 2007 and December 2008. MATERIALS： Dingnaofang consisted of Huangqi （Milkvetch Root; Radix Astragah）, Chuanxiong （Szechwan Lovage Rhizome; Rhizoma Chuanxiong）, Yinxingye （ginkgo leaf; Fofium Ginkgo）, Dilong （earthworm; Pheretima）, Danggui （Chinese Angelica; Radix Angelicae Sinensis）, Tianqi （Radix Notoginseng）, and Gancao （Radix Glycyrrhizae; Radix Glycytthizae）, with a proportion of 5：2：2： 1： 1： 1： 1. METHODS： A total of 130 Sprague Dawley rats were randomly divided into sham-surgery （n = 10）, cerebral ischemia/reperfusion （n = 40）, cerebral ischemia pretreatment （n = 40） and Dingnaofang pretreatment groups （n = 40）. Middle cerebral artery occlusion was used to induce cerebral ischemic injury. The bilateral common carotid artery in the cerebral ischemia pretreatment group was blocked for 10 minutes on days 7, 3 and 1 prior to ischemia/reperfusion injury, while rats in Dingnaofang pretreatment group were intragastrically administrated with 4 g Dingnaofang 1 week prior to cerebral ischemia once per day, for 7 successive days. MAIN OUTCOME MEASURES： Apoptosis ratios in vascular endothelial cells were measured using Hoechst 33258 staining and flow cytometry; apoptosis was detected by monitoring DNA gradient bands and the activation of caspase-3, 8, 9 and Bid using Western blot. RESULTS： Following cerebral ischemiaJreperfusion injury, the number of apoptotic vascular endothelial cells in the middle cerebral artery significantly increased （P 〈 0.01）; however, cerebral ischemia pretreatment and Dingnaofang pretreatment groups significantly reduced apoptosis induced by cerebral ischemia/reperfusion injury （P 〈 0.01）. In particular, DNA gradient bands were not observed following Dingnaofang pretreatment. At 24 hours after cerebral ischemia/reperfusion injury, cleaved fragments of caspase-3, 8 and 9 were detected at 11 kD （P 11）, 20 kD （P 20） and 10 kD （P 10）, respectively, following Western blot. Bid was also cleaved into its truncated form （tBid; 15 kD）. Gray scale analysis indicated that P 11, P 20, P 10 and tBid band values in the Dingnaofang pretreatment group were significantly less than in the cerebral ischemia/reperfusion and cerebral ischemia pretreatment groups （P 〈 0.01 or P 〈 0.05）. CONCLUSION： Dingnaofang inhibits vascular endothelial cell apoptosis induced by cerebral ischemia/reperfusion injury via inhibition of apoptotic signal transduction pathways.
基金supported by the National Natural Science Foundation of China,Nos.82172196(to KX),82372507(to KX)the Natural Science Foundation of Hunan Province,China,No.2023JJ40804(to QZ)the Key Laboratory of Emergency and Trauma(Hainan Medical University)of the Ministry of Education,China,No.KLET-202210(to QZ)。
文摘Ischemia–reperfusion injury is a common pathophysiological mechanism in retinal degeneration.PANoptosis is a newly defined integral form of regulated cell death that combines the key features of pyroptosis,apoptosis,and necroptosis.Oligomerization of mitochondrial voltage-dependent anion channel 1 is an important pathological event in regulating cell death in retinal ischemia–reperfusion injury.However,its role in PANoptosis remains largely unknown.In this study,we demonstrated that voltage-dependent anion channel 1 oligomerization-mediated mitochondrial dysfunction was associated with PANoptosis in retinal ischemia–reperfusion injury.Inhibition of voltage-dependent anion channel 1 oligomerization suppressed mitochondrial dysfunction and PANoptosis in retinal cells subjected to ischemia–reperfusion injury.Mechanistically,mitochondria-derived reactive oxygen species played a central role in the voltagedependent anion channel 1-mediated regulation of PANoptosis by promoting PANoptosome assembly.Moreover,inhibiting voltage-dependent anion channel 1 oligomerization protected against PANoptosis in the retinas of rats subjected to ischemia–reperfusion injury.Overall,our findings reveal the critical role of voltage-dependent anion channel 1 oligomerization in regulating PANoptosis in retinal ischemia–reperfusion injury,highlighting voltage-dependent anion channel 1 as a promising therapeutic target.
基金supported by the National Natural Science Foundation of China,Nos.82260245(to YX),81660207(to YX),81960253(to YL),82160268(to YL),U1812403(to ZG)Science and Technology Projects of Guizhou Province,Nos.[2019]1440(to YX),[2020]1Z067(to WH)+1 种基金Cultivation Foundation of Guizhou Medical University,No.[20NSP069](to YX)Excellent Young Talents Plan of Guizhou Medical University,No.(2022)101(to WH)。
文摘Several studies have shown that activation of unfolded protein response and endoplasmic reticulum(ER)stress plays a crucial role in severe cerebral ischemia/reperfusion injury.Autophagy occurs within hours after cerebral ischemia,but the relationship between ER stress and autophagy remains unclear.In this study,we established experimental models using oxygen-glucose deprivation/reoxygenation in PC12 cells and primary neurons to simulate cerebral ischemia/reperfusion injury.We found that prolongation of oxygen-glucose deprivation activated the ER stress pathway protein kinase-like endoplasmic reticulum kinase(PERK)/eukaryotic translation initiation factor 2 subunit alpha(e IF2α)-activating transcription factor 4(ATF4)-C/EBP homologous protein(CHOP),increased neuronal apoptosis,and induced autophagy.Furthermore,inhibition of ER stress using inhibitors or by si RNA knockdown of the PERK gene significantly attenuated excessive autophagy and neuronal apoptosis,indicating an interaction between autophagy and ER stress and suggesting PERK as an essential target for regulating autophagy.Blocking autophagy with chloroquine exacerbated ER stress-induced apoptosis,indicating that normal levels of autophagy play a protective role in neuronal injury following cerebral ischemia/reperfusion injury.Findings from this study indicate that cerebral ischemia/reperfusion injury can trigger neuronal ER stress and promote autophagy,and suggest that PERK is a possible target for inhibiting excessive autophagy in cerebral ischemia/reperfusion injury.
文摘Nitric oxide(NO)is a gaseous molecule produced by 3 different NO synthase(NOS)isoforms:Neural/brain NOS(nNOS/bNOS,type 1),endothelial NOS(eNOS,type 3)and inducible NOS(type 2).Type 1 and 3 NOS are constitutively expressed.NO can serve different purposes:As a vasoactive molecule,as a neurotransmitter or as an immunomodulator.It plays a key role in cerebral ischemia/reperfusion injury(CIRI).Hypoxic episodes simulate the production of oxygen free radicals,leading to mitochondrial and phospholipid damage.Upon reperfusion,increased levels of oxygen trigger oxide synthases;whose products are associated with neuronal damage by promoting lipid peroxidation,nitrosylation and excitotoxicity.Molecular pathways in CIRI can be altered by NOS.Neuroprotective effects are observed with eNOS activity.While nNOS interplay is prone to endothelial inflammation,oxidative stress and apoptosis.Therefore,nNOS appears to be detrimental.The interaction between NO and other free radicals develops peroxynitrite;which is a cytotoxic agent.It plays a main role in the likelihood of hemorrhagic events by tissue plasminogen activator(t-PA).Peroxynitrite scavengers are currently being studied as potential targets to prevent hemorrhagic transformation in CIRI.
基金Shanghai Rehabilitation Medical Association,Grant/Award Number:2023JGKT24China Rehabilitation Medical Association,Grant/Award Number:KFKT-2023Shanghai“14th Five-Year Plan”Traditional Chinese Medicine Specialty and Traditional Chinese Medicine Emergency Capacity Improvement Project,Grant/Award Number:ZYTSZK2-7。
文摘Cerebral ischemia/reperfusion(I/R)injury is an important pathophysiological condition of ischemic stroke that involves a variety of physiological and pathological cell death pathways,including autophagy,apoptosis,necroptosis,and phagoptosis,among which autophagy is the most studied.We have reviewed studies published in the past 5 years regarding the association between autophagy and cerebral I/R injury.To the best of our knowledge,this is the first review article summarizing potential candidates targeting autophagic pathways in the treatment of I/R injury post ischemic stroke.The findings of this review may help to better understand the pathogenesis and mechanisms of I/R events and bridge the gap between basic and translational research that may lead to the development of novel therapeutic approaches for I/R injury.
基金Key Science and Technology Program of Shaanxi Province,Grant/Award Number:2024SF2-GJHX-45National Natural Science Foundation of China,Grant/Award Number:82472191The Natural Science Foundation of Shaanxi Province,Grant/Award Number:2024JC-ZDXM-49。
文摘Background:Ex vivo lung perfusion(EVLP)has emerged as a critical technique for lung preservation and evaluation prior to transplantation.While conventional rat EVLP systems utilize closed-loop dual cannulation of pulmonary artery(PA)and vein,the effect of the simplified model using single PA cannulation with passive venous drainage is unknown.Methods:We developed two EVLP models in rats:a semi-closed circuit with PA-only cannulation and left atrial incision for passive venous drainage(SC-EVLP),and a closed circuit employing both arterial and venous cannulation(C-EVLP).Donor lungs were perfused for a defined duration and subsequently orthotopically transplanted.We evaluated pulmonary function parameters,histopathological injury scores,inflammatory cytokine levels,and apoptotic marker expression at the end of perfusion and posttransplantation.Results:Compared to the conventional EVLP,the SC-EVLP group exhibited significantly lower PA pressure and improved dynamic lung compliance throughout perfusion.Although the levels of tumor necrosis factor-αin the perfusate were higher in the SC-EVLP group,other cytokine levels in the perfusate and bronchoalveolar lavage fluid exhibited no significant differences.Pulmonary edema was reduced in the SC-EVLP group,as indicated by a lower lung wet-to-dry ratio.After transplantation,lungs from the SC-EVLP group exhibited lower histological injury scores,reduced apoptosis,and decreased serum cytokine levels,suggesting attenuated inflammation and tissue damage.Conclusions:In a rat model,single PA cannulation with passive venous drainage reduced pulmonary edema during EVLP and reduced lung injury and systemic inflammation after transplantation.
基金sponsored by the National Natural Science Foundation of China(Nos.82027802,82101389,82274401,and 81971114)Beijing Nova Program(No.20230484286)+1 种基金Beijing Natural Science Foundation(7254366)the General Project of Science and Technology of Beijing Municipal Education Commission(No.KM202110025018).
文摘Objective Cerebral venous outflow disorders(CVOD)can impair cerebral perfusion and produce diverse,often debilitating symptoms,substantially reducing quality of life.Intermittent hypoxiahyperoxia training(IHHT)has demonstrated therapeutic potential across various pathologies and may represent a promising non-pharmacological approach for CVOD management.Methods Patients with imaging-confirmed CVOD underwent 14 IHHT sessions,each comprising four cycles of 10-minute hypoxia(11%O_(2))stimulation and 20-minute hyperoxia(38%O_(2)).Physiological parameters and adverse events were monitored throughout the intervention.Clinical scales,24-hour ambulatory blood pressure,blood tests,jugular ultrasound,and perfusion imaging were assessed preand post-intervention.Results No participants experienced intolerable discomfort or severe adverse events;vital signs remained within normal ranges.No significant changes were observed in 24-hour blood pressure,blood cell counts,lipid profiles,or other blood markers.Notably,60%of patients(n=12)reported overall symptom improvement on the Patient Global Impression of Change scale.Headache severity,as measured by the visual analogue scale,significantly decreased(6.33±1.22 vs.4.89±2.03,P=0.016).In patients with internal jugular vein(IJV)stenosis,significant improvements were observed in regional cerebral blood flow(including the insula,occipital lobe,internal capsule,and lenticula)and left J3-segment IJV flow volume(107.27[47.50,160.00]vs.140.83[55.00,210.00]mL/min,P=0.011).Conclusion The current IHHT protocol is safe and well-tolerated in patients with CVOD.IHHT may alleviate CVOD-related symptoms by improving oxygen saturation,cerebral perfusion,and venous outflow pattern,supporting its potential as a non-invasive therapeutic strategy.
基金supported by the National Natural Science Foundation of China(81770692)。
文摘Objective:Acute kidney injury(AKI)is a clinical syndrome characterized by sudden deterioration of renal function,with ischemia reperfusion injury(IRI)being the most common cause.Long noncoding RNA(lncRNA)regulate cell fate through interactions with microRNA(miRNA)and messenger RNAs(mRNA),but the mechanisms and regulatory networks underlying lncRNA AK154753(AK154753)in IRI-induced AKI remain unclear.This study aims to investigate the role of AK154753 in acute renal IRI and to elucidate the molecular mechanism of the AK154753 via miR-345-3p/Bcl-2 homologous antagonist/killer(Bak)and miR-708-5p/Bcl-2 interacting mediator of cell death(Bim)axis.Methods:A bilateral renal artery ischemia model was established in mice(30 minutes ischemia followed by 24 hours and 48 hours reperfusion).Kidney tissues were analyzed using microarray-based transcriptomic sequencing to identify differentially expressed lncRNAs,miRNAs,and mRNAs.RNA levels of AK154753,miR-345-3p,miR-708-5p,Bak,and Bim were validated using real-time reverse transcription PCR(real-time RTPCR).Oxygen and glucose deprivation/reperfusion(OGD/R)models were constructed in mouse proximal renal tubular epithelial BUMPT cells to simulate in vitro IRI conditions.Adeno-associated virus(AAV)-mediated shRNA was used to silence AK154753 in vivo.Apoptosis was assessed using TUNEL staining and flow cytometry.Protein levels of Bak,Bim,and cleaved-caspase-3 were measured using Western blotting.Fluorescence in situ hybridization(FISH)was used to determine intracellular localization of AK154753.Binding relationships between AK154753 and miR-345-3p/Bak and miR-708-5p/Bim were verified using dual-luciferase reporter assays.MiRNA mimics and inhibitors were used to evaluate regulatory-network integrity.Results:IRI significantly elevated serum blood urea nitrogen(BUN)and serum creatinine(Scr),accompanied by tubular-structure damage and increased cell apoptosis(all P<0.05).Transcriptome profiling and real-time RT-PCR validation demonstrated that lncRNA AK154753,along with the pro-apoptotic proteins Bak and Bim,was significantly upregulated after IRI,whereas miR-345-3p and miR-708-5p were markedly downregulated(P<0.01).In vitro,OGD/R treatment significantly induced AK154753 expression in renal tubular epithelial cells and suppressed the expression of miR-345-3p and miR-708-5p,while markedly increasing the protein levels of Bak,Bim,and cleaved-caspase 3,resulting in a significant increase in apoptosis(all P<0.01).Silencing AK154753 significantly attenuated OGD/R-induced apoptosis,reduced the expression of Bak,Bim,and cleaved caspase 3,and decreased cell apoptosis(all P<0.01),while significantly upregulating miR-345-3p and miR-708-5p expression(P<0.01).In vivo,adeno-associated virus(AAV)-mediated knockdown of AK154753 significantly improved renal function in IRI mice,alleviated tubular injury,and suppressed renal tissue apoptosis,as evidenced by reduced BUN and Scr levels,improved histopathological injury scores,and decreased expression of Bak,Bim,and cleaved caspase-3(all P<0.01),accompanied by significant upregulation of miR-345-3p and miR-708-5p(all P<0.01).Luciferase reporter assays further confirmed that miR-345-3p directly binds to the 3'-untranslated region(3'-UTR)of AK154753 and Bak,whereas miR-708-5p directly binds to the 3'-UTRs of AK154753 and Bim.Inhibition of miR-345-3p or miR-708-5p abolished the anti-apoptotic effects induced by AK154753 silencing and restored Bak and Bim expression levels(all P<0.01).Conclusion:AK154753 is upregulated in acute renal IRI and promotes apoptosis by suppressing miR-345-3p and miR-708-5p,thereby upregulating Bak and Bim,and participates in the initiation and progression of acute renal IRI.
文摘Introduction: Renal ischemia-reperfusion (IR) is responsible for injuries such as destruction or dysfunction of tubular epithelial cells with inflammatory reaction and oxidative stress. Several therapeutic methods have been tested to alleviate ischemia-perfusion injury, ranging from using anti-inflammatory drugs, antioxidants, and plants from traditional pharmacopeia to administering RNA interference. However, there is currently no effective therapeutic option available for the treatment of renal IR injury, other than supportive therapies such as renal replacement therapy or hydration. Objective: This present study aimed to evaluate the effect of Guiera senegalensis on renal ischemia reperfusion, a recognized plant for its antioxidant and anti-inflammatory properties. Materials and Methods: Twenty-four (24) adult male Wistar rats were divided into four following groups: SLAM (subjected to a median laparotomy with simulated ischemia);GUIERRA (animals that received 250 mg/kg of guierra senegalensis orally, once a day, for 5 days, with simulated renal ischemia);IR (animals that underwent laparotomy followed by clamping of bilateral renal pedicles for 45 min and followed by reperfusion);GUIERRA + IR (animals given GUIERRA at the dosage of 250 mg/kg per day, for 5 days and then subjected to renal ischemia-reperfusion). Data analysis was performed by ANOVA, and a significance level of p Results: Compared with the I/R group, rats in the GUIERRA + IR group showed reduced histopathological damage scores (p Conclusion: The results of this preliminary work suggest that Guiera senegalensis decreases the degree of tissue damage in renal ischemia-reperfusion cases. This plant seems to be a promising therapeutic;further studies could help to precise the targets of its compounds on ischemia-reperfusion pathways.
基金supported by the Natural Science Foundation of Shandong Province,No.ZR2023MC168the National Natural Science Foundation of China,No.31670989the Key R&D Program of Shandong Province,No.2019GSF107037(all to CS).
文摘Vascular endothelial growth factor and its mimic peptide KLTWQELYQLKYKGI(QK)are widely used as the most potent angiogenic factors for the treatment of multiple ischemic diseases.However,conventional topical drug delivery often results in a burst release of the drug,leading to transient retention(inefficacy)and undesirable diffusion(toxicity)in vivo.Therefore,a drug delivery system that responds to changes in the microenvironment of tissue regeneration and controls vascular endothelial growth factor release is crucial to improve the treatment of ischemic stroke.Matrix metalloproteinase-2(MMP-2)is gradually upregulated after cerebral ischemia.Herein,vascular endothelial growth factor mimic peptide QK was self-assembled with MMP-2-cleaved peptide PLGLAG(TIMP)and customizable peptide amphiphilic(PA)molecules to construct nanofiber hydrogel PA-TIMP-QK.PA-TIMP-QK was found to control the delivery of QK by MMP-2 upregulation after cerebral ischemia/reperfusion and had a similar biological activity with vascular endothelial growth factor in vitro.The results indicated that PA-TIMP-QK promoted neuronal survival,restored local blood circulation,reduced blood-brain barrier permeability,and restored motor function.These findings suggest that the self-assembling nanofiber hydrogel PA-TIMP-QK may provide an intelligent drug delivery system that responds to the microenvironment and promotes regeneration and repair after cerebral ischemia/reperfusion injury.
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.
文摘Objective:To evaluate the cardioprotective effects of sinomenine using the ischemia/reperfusion(I/R)rat model.Methods:Wistar rats were randomly divided into 6 groups:group Ⅰ with reperfusion,group Ⅱ perfused with sinomenine,group Ⅲ perfused with 5-hydroxydecanoate(5-HD),group Ⅳ perfused with 5-HD+sinomenine,group Ⅴ perfused with L-nitro arginine methyl ester(L-NAME),group Ⅵ perfused with L-NAME+sinomenine.Myocardial ischemia was induced by interrupting the aortic blood supply for 30 min,followed by reperfusion(55 min).Cardiac,hepatic,antioxidant,and inflammatory parameters were assessed.Additionally,endothelin,tissue factor,platelet-activating factor,plasminogen activator inhibitor,plasma fibrinogen,and thromboxane B2 were also analyzed.Results:Administration of 5-HD or L-NAME,used as the selective antagonist of mitoKATP and NO system,respectively,resulted in significantly increased levels of premature ventricular complexes,lactate dehydrogenase,ventricular fibrillation,ventricular tachycardia,and arrhythmia intensity(P<0.05).In contrast,sinomenine significantly reduced the level of troponin Ⅰ,lactate dehydrogenase,creatine kinase,and creatine kinase MB compared to the 5-HD group and the L-NAME group(P<0.05).Additionally,sinomenine significantly reduced malondialdehyde level and enhanced the levels of superoxide dismutase,glutathione peroxidase,catalase,and glutathione/glutathione disulfide ratio(P<0.05).It also significantly suppressed the levels of endothelin-1,platelet-activating factor,tissue factor,plasminogen activator inhibitor 1,thromboxane B2,and plasma fibrinogen(P<0.05).Conclusions:These results suggest that sinomenine exhibits significant cardioprotection effects against I/R-induced cardiac injury in rats.
基金supported by 2024 Research Grant from Kangwon National University(M.C.S)and the Basic Science Research Program through the National Research Foundation of Korea(NRF)funded by the Ministry of Education(NRF-2021R1A2C1094224)(J.H.A).
文摘Background:Astrocyte endfeet(AEF)serves as a key element of the blood-brain barrier and is important for the survival and maintenance of neuronal function.However,the immunohistochemical and ultrastructural changes of AEF in the CA1 and CA3 areas of the hippocampus over time following cerebral ischemia-reperfusion(IR)injury have not been well elucidated.Objectives:We investigated chronological changes in AEF in the gerbil hippocampal CA1 area from 3 h to 10 days following transient forebrain ischemia(TFI),and examined their association with neuronal death and tissue repair following IR injury.Changes in the CA3 area were also examined at 10 days post-TFI for comparative purposes.Methods:Neuronal death was confirmed using histochemistry,immunohistochemistry,and histofluorescence.Changes in AEF were examined by double immunofluorescence with glial fibrillary acidic protein(GFAP)and glucose transporter 1(GLUT1),and by transmission electron microscopy(TEM)for ultrastructural changes.Results:Significant TFI-induced neuronal death occurred in the CA1 area on day 5 following IR injury and persisted until 10 days after TFI,while no neuronal death(or loss)was found in the CA3 area after TFI.Looking at TFI-induced changes in AEF,at 3 and 6 h after TFI,GFAP-immunoreactive(+)AEF in the CA1 area appeared swollen and harbored enlarged,dark mitochondria,and the swelling was reduced by 1-day post-TFI.On 2 and 5 days following TFI,GFAP+AEF were markedly enlarged and fragmented,containing shrunken mitochondria,vacuolations,and sparse organelles.Ten days post-TFI,the ends of GFAP+astrocytic processes extended to microvessels,appeared edematous,and were filled with cellular debris.In the CA3 area,AEF was slightly dilated at 10 days after TFI.These findings indicate that damage to or disruption of AEF in the CA1 area occurs in the early phase after 5-min TFI but is rarely observed in the CA3 area.Conclusion:Taken together,damage to or disruption of AEF following ischemic insults may be strongly linked to neuronal death/loss.
基金Supported by Zhejiang Provincial Natural Science Foundation of China,No.LQ23H020004The Medical and Health Research Project of Zhejiang province,No.2024KY983Basic Medical Health Technology Project of Wenzhou Science and Technology Bureau,No.Y20210818 and No.Y20210140.
文摘BACKGROUND Myocardial ischemia/reperfusion(I/R)injury,which is associated with high morbidity and mortality,is a main cause of unexpected myocardial injury after acute myocardial infarction.However,the underlying mechanism remains unclear.Circular RNAs(circRNAs),which are formed from protein-coding genes,can sequester microRNAs or proteins,modulate transcription and interfere with splicing.Authoritative studies suggest that circRNAs may play an important role in myocardial I/R injury.AIM To explore the role and mechanism of circRNAs in myocardial I/R injury.METHODS We constructed a myocardial I/R injury model using ligation of the left anterior descending coronary artery,and evaluated the success of the validated model using triphenyltetrazolium chloride and hematoxylin-eosin staining.Then,left ventricular samples from different groups were selected for mRNA-sequence,and differential gene screening was performed on the obtained results.The differentially obtained mRNAs were divided into up-regulated and down-regulated according to their expression levels,and Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)functional enrichment analysis were performed,respectively.Then,the obtained circRNA and microRNA(miRNA)were paired for analysis,and the binding sites of miRNA and mRNA were virtual screened.Finally,the obtained circRNA,miRNA and mRNA were constructed by ceRNA mutual most useful network.RESULTS We used an RNA sequencing array to investigate the expression signatures of circRNAs in myocardial I/R injury using three samples from the I/R group and three samples from the sham group.A total of 142 upregulated and 121 downregulated circRNAs were found to be differentially expressed(fold change≥2,P<0.05).GO and KEGG functional analyses of these circRNAs were performed.GO analysis revealed that these circRNAs were involved mainly in cellular and intracellular processes.KEGG analysis demonstrated that 6 of the top 20 pathways were correlated with cell apoptosis.Furthermore,a circRNA-miRNA coexpression network and ceRNA network based on these genes were constructed,revealing that mmu-circ-0001452,mmu-circ-0001637,and mmu-circ-0000870 might be key regulators of myocardial I/R injury.CONCLUSION This research provides new insights into the mechanism of myocardial I/R,which mmu-circ-0001452,mmu-circ-0001637,and mmu-circ-0000870 are expected to be new therapeutic targets for myocardial I/R injury.
基金supported by a grant from the Russian Science Foundation(23-75-01061)。
文摘Objectives:Skeletal muscle ischemia/reperfusion injury(IRI)occurs as a result of a marked reduction in arterial perfusion to a limb and can lead to tissue death and threaten limb viability.This work assessed the effects of 20-hydroxyecdysone(20E)on hindlimb skeletal tissue following tourniquet-induced ischemia/reperfusion injury.Methods:Animals were divided into 4 groups—control group(Control),Control+20E(C+20E),mice with IRI(IRI),and mice with IRI+20E(IRI+20E).IRI was modeled by applying a tourniquet to the hind limb for 2 h with reperfusion for 1 h.5 mg/kg of 20E was administered intraperitoneally for 14 days.Afterward,the physical activity of mice,the histological structure of the quadriceps femoris,the expression of genes encoding proteins induced by hypoxia and involved in tissue adaptation to ischemia,and the functional parameters of skeletal muscle mitochondria were assessed.Results:It was shown that IRI of the limbs leads to functional disorders,depression of muscle function,accumulation of malondialdehyde(MDA)in mitochondria,and a decrease in their Ca2+buffering capacity,as well as an increase in the expression of HIF-1α,VGEF-A,PGC1αand PDGF-BB genes associated with adaptation to ischemia.20E reduced the intensity of degenerative processes in skeletal muscles,which was expressed in a decrease in the number of centrally nucleated fibers.Analysis of gene expression levels indicated a high degree of adaptation of animals to IRI.20E reduced the level of MDA in mitochondria,but did not affect the rate of respiration and calcium retention capacity of organelles both in normal conditions and during IRI.Conclusion:20E partially alleviates the skeletal muscle damage caused by IRI and can be used as part of combination therapy.
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 for Distinguished Young Scholars(No.81725021)the National Natural Science Foundation of China(Nos.82003633 and 82173705)。
文摘Hepatic ischemia/reperfusion injury(IRI)remains a critical complication contributing to graft dysfunction following liver surgery.As part of an ongoing search for hepatoprotective natural products,five previously unreported homoadamantane-type polycyclic polyprenylated acylphloroglucinols(PPAPs),named hyperhomanoons A-E(1-5),and one known analog,hypersampsone O(6),were isolated from Hypericum patulum.Among these,compound 6 demonstrated potent protective effects against CoCl_(2)-induced hypoxic injury in hepatocytes.Furthermore,in a murine model of hepatic IRI induced by vascular occlusion,pretreatment with 6 markedly alleviated liver damage and reduced hepatocyte apoptosis.This study is the first to identify PPAPs as promising scaffolds for the development of therapeutic agents targeting hepatic IRI,underscoring their potential as lead compounds in drug discovery efforts for ischemic liver diseases.
文摘Increasing evidence of the significant clinical value of protection against ischemia/reperfusion injury has contributed to the realization of the independent importance of this approach in improving prognosis and reducing cardiovascular mortality.Extracellular vesicles(EVs)derived by adipose mesenchymal stem cells may mediate the paracrine effects of stem cells and provide regenerative and anti-inflammatory properties,which are enhanced byγ-aminobutyric acid.The protective effects on cardiac myocytes may result from the EV embarked by miR-21-5p,which is a target for thioredoxin-interacting protein,regulating the formation of thioredoxin-interacting protein-thioredoxin complexes and subsequently enhancing the antioxidant activity of thioredoxin.It has been found thatγ-aminobutyric acid governs myocardial bioenergetics through suppressing inflammation and supporting mitochondrial structure.Finally,stem cell-based cell-free therapy based on adipose-derived stem cell EVs is considered a promising approach to individualized management of ischemia-induced cardiomyopathy.
基金supported by the National Natural Science Foundation of China(81473453,81673800)the Projects of International Science and Technology Cooperation in Henan(182102410084).
文摘Objective MicroRNA-1(miR-1)aggravates myocardial ischemia–reperfusion(I/R)injury,whereas insulin-like growth factor-1(IGF-1)maintains cardiomyocyte homeostasis.In this study,the aim is to investigate whether miR-1 can exacerbate I/R injury through the regulation of IGF-1.Methods The infarct area,lactate dehydrogenase,miR-1 level,and apoptosis level were examined in the Langendorff isolated rat I/R model.The hypoxia–reoxygenation model of rat cardiacmyocytes and H9c2 cells were developed to determine the levels of miR-1,IGF-1 mRNA,and IGF-1 protein.Furthermore,the dual-luciferase assay was used to verify the relationship between miR-1 and IGF-1.Results Overexpression of miR-1 increased the level of apoptosis and decreased the IGF-1 expression.However,inhibition of miR-1 expression decreased the level of apoptosis,alleviated the degree of injury,and increased the IGF-1 expression.Overexpression of IGF-1 also reduced the degree of cellular damage and level of apoptosis caused by the overexpression of miR-1.When IGF-1 was knocked down,myocardial cells displayed more severe damage and a higher apoptosis level,even with decreased levels of miR-1.Conclusion miR-1 promotes apoptosis and aggravates I/R injury by downregulating IGF-1.
文摘BACKGROUND Hepatic ischemia reperfusion(HIR)injury is a major complication affecting various major liver surgeries,including liver transplantation.Aprepitant(APRE),a neurokinin-1 receptor antagonist,is commonly used as an antiemetic to prevent chemotherapy-induced nausea and vomiting.AIM To assess the potential protective effect of APRE against HIR-induced liver injury via targeting the nucleotide-binding oligomerization domain-,leucine-rich repeat-,and pyrin domain-containing receptor 3/interleukin(IL)-1beta signaling pathway.METHODS Six groups of adult male Wistar albino rats were divided as follows:Sham group,Sham/APRE10 group(APRE 10 mg/kg),HIR group,HIR/APRE5 group(APRE 5 mg/kg),HIR/APRE10 group(APRE 10 mg/kg),and HIR/APRE20 group(APRE 20 mg/kg).Serum alanine transaminase,aspartate transaminase,liver malondialdehyde,total antioxidant capacity levels,as well as IL-6,sirtuin 1(Sirt1),caspase-3,cleaved caspase-3,and tumor necrosis factor alpha biomarkers,were evaluated.Hepatic specimens were examined histopathologically and immunohistochemically for nuclear factor erythroid-2-related factor 2(Nrf2)immunoexpression.RESULTS HIR resulted in hepatic damage,as evidenced by histopathological changes and a significant increase in serum alanine transaminase,aspartate transaminase,hepatic malondialdehyde,caspase-3,and tumor necrosis factor alpha levels.Additionally,there were significant increases in hepatic total antioxidant capacity and reductions in IL-6 and cleaved caspase-3 protein levels,as demonstrated by Western blot analysis,along with enhanced immunoexpression of Sirt1 and Nrf2.APRE has significantly reduced various parameters of oxidative stress,inflammation,and apoptosis,and a significant increase in liver Nrf2 immunoexpression,leading to a significant improvement in the histopathological changes.CONCLUSION In conclusion,targeting the Sirt1/Nrf2 signaling pathway,as demonstrated by APRE in our model,could present a promising therapeutic target to protect against HIR-induced liver injury during major liver surgeries.
