Hypoxia is one of the major environmental stressors,frequently resulting in serious yield losses for maricultured large yellow croaker(Larimichthys crocea).We know that pyroptosis contributes to tissue damage under hy...Hypoxia is one of the major environmental stressors,frequently resulting in serious yield losses for maricultured large yellow croaker(Larimichthys crocea).We know that pyroptosis contributes to tissue damage under hypoxic conditions.However,whether GSDMEs-mediated pyroptosis is involved in hypoxia-induced tissue damage in fish remains unclear.In the present study,two Lcgsdme genes,Lcgsdmea/b,were cloned and characterized from the large yellow croaker.Both genes contain a conserved N-terminalpore-forming domain,a C-terminal auto-inhibitory domain,and a flexible hinge region.After hypoxia stress,the expression of Lcgsdmea/b transcripts and proteins in the liver were significantly higher than in unstressed fish.The proteins of LcGSDMEa/b could be cleaved under hypoxic conditions.Compared to LcGSDMEb,the expression of LeGSDMEa was higher in both mRNA and protein levels,thus exhibiting a stronger response during hypoxia stress.Furthermore,after 48 h of hypoxia stress,approximately 65%liver cells exhibited abnormalities,with pyroptosis being detected using a transmission electron microscope.TUNEI/LeGSDMEa double staining assay revealed a high expression of LcGSDMEa in the dead cells.We observed a significant up-regulation of pyroptosis pathway genes(asc,caspase-3)and pro-inflammatory cytokine genes(il-1β,il-18).After simultaneous knockdown of Legsdmea/b in vivo,the liver exhibited better health compared to the control group,with less cell swelling and vacuolation.Taken together,these findings demonstrate that hypoxia stress could activate LcGSDMEa/b and induce pyroptosis in the liver of large yellow croakers,thereby contributing to tissue damage.Our study improves the understanding of hypoxia-induced tissue damage in fish,and provides new clues for protecting fish against hypoxia-induced damage.展开更多
Microglia,the resident monocyte of the central nervous system,play a crucial role in the response to spinal cord injury.However,the precise mechanism remains unclear.To investigate the molecular mechanisms by which mi...Microglia,the resident monocyte of the central nervous system,play a crucial role in the response to spinal cord injury.However,the precise mechanism remains unclear.To investigate the molecular mechanisms by which microglia regulate the neuroinflammatory response to spinal cord injury,we performed single-cell RNA sequencing dataset analysis,focusing on changes in microglial subpopulations.We found that the MG1 subpopulation emerged in the acute/subacute phase of spinal cord injury and expressed genes related to cell pyroptosis,sphingomyelin metabolism,and neuroinflammation at high levels.Subsequently,we established a mouse model of contusive injury and performed intrathecal injection of siRNA and molecular inhibitors to validate the role of ceramide synthase 5 in the neuroinflammatory responses and pyroptosis after spinal cord injury.Finally,we established a PC12-BV2 cell co-culture system and found that ceramide synthase 5 and pyroptosis-associated proteins were highly expressed to induce the apoptosis of neuron cells.Inhibiting ceramide synthase 5 expression in a mouse model of spinal cord injury effectively reduced pyroptosis.Furthermore,ceramide synthase 5-induced pyroptosis was dependent on activation of the NLRP3 signaling pathway.Inhibiting ceramide synthase 5 expression in microglia in vivo reduced neuronal apoptosis and promoted recovery of neurological function.Pla2g7 formed a“bridge”between sphingolipid metabolism and ceramide synthase 5-mediated cell death by inhibiting the NLRP3 signaling pathway.Collectively,these findings suggest that inhibiting ceramide synthase 5 expression in microglia after spinal cord injury effectively suppressed microglial pyroptosis mediated by NLRP3,thereby exerting neuroprotective effects.展开更多
Background:Cisplatin triggers Gasdermin E(GSDME)cleavage,causing membrane bubble formation,content release,and inflammation.Caspase-3 activation initiates GSDME cleavage,and thus inhibiting this pathway mitigates cisp...Background:Cisplatin triggers Gasdermin E(GSDME)cleavage,causing membrane bubble formation,content release,and inflammation.Caspase-3 activation initiates GSDME cleavage,and thus inhibiting this pathway mitigates cisplatin-induced pyroptosis in hepatocytes.This study aimed to delve into how cisplatin induces liver injury via pyroptosis.Methods:For animal experiments,C57BL/6J mice were divided into three groups:control,liver injury model group,and Ac-DMLD-CMK(caspase-3 inhibitor)intervention group.The liver histology was evaluated by hematoxylin and eosin staining,immunohistochemistry,immunofluorescence and TUNEL staining.The mRNA and protein levels were detected by real-time polymerase chain reaction(PCR)and Western blot analysis.For in vitro experiments,HL-7702 cells were treated with cisplatin or GSDME siRNA.Cell pyroptosis was determined via cellular morphology,cytotoxicity and viability detection,flow cytometric assay,and Western blot detection for the expression of pyroptosis-related proteins.Results:Cisplatin-induced distinct liver morphological changes,hepatocellular injury,and inflammation in mice,along with elevated serum alanine aminotransferase(ALT)and aspartate aminotransferase(AST)levels and increased pro-inflammatory cytokine expression.Heightened macrophage infiltration and hepatocellular death indicated cisplatin-induced hepatotoxicity.Cisplatin upregulated GSDME activation,along with Bax-mediated caspase-3 cleavage both in vivo and in vitro,implicating caspase-3/GSDME-dependent pyroptosis in liver injury.Treatment with Ac-DMLD-CMK ameliorated cisplatin-induced liver injury,reducing hepatocellular lesions,serum ALT and AST levels,cytokine expression,macrophage infiltration,and hepatocyte death.Ac-DMLD-CMK also attenuated GSDME-dependent pyroptosis post-cisplatin induction,as evidenced by decreased GSDME expression,Bax upregulation,and cleaved caspase-3 activation.For HL-7702 cells,GSDME siRNA transfection reduced GSDME expression,attenuated typical signs of cisplatin-induced pyroptosis,partially restored cell viability,and significantly inhibited cytotoxicity and a decrease in the proportion of propidium iodide-positive cells,indicating protection against cisplatininduced hepatocyte pyroptosis.Conclusions:Our study underscores the role of the caspase-3/GSDME signaling pathway in mediating cisplatin-induced hepatotoxicity,particularly in cases of excessive or cumulative cisplatin exposure.These findings suggest that targeting GSDME could represent a promising therapeutic approach to mitigate cisplatin-induced liver damage.展开更多
BACKGROUND Damage associated molecular patterns(DAMPs)are vital for the immunogenic cell death of cancer cells and can enhance the anti-tumor activity of immune cells in colorectal cancer(CRC).Peroxiredoxin 1(Prdx1),a...BACKGROUND Damage associated molecular patterns(DAMPs)are vital for the immunogenic cell death of cancer cells and can enhance the anti-tumor activity of immune cells in colorectal cancer(CRC).Peroxiredoxin 1(Prdx1),an important DAMP,is highly expressed in various tumor tissues including CRC.However,the role of Prdx1 in CRC remains unknown.AIM To investigate the effect and mechanisms of Prdx1 on CRC.METHODS Patients diagnosed with CRC in our medical center were included in this study to verify the expression of Prdx1 in cancer tissues.Recombinant Prdx1(rPrdx1)was used to stimulate RKO and SW480 colon cancer cells.The cell survival rate,migration,proliferation and invasion ability were assessed.Transmission electron microscopy,TUNEL assay,lactate dehydrogenase release assay,and Western blot were used to determine the effect of Prdx1 on pyroptosis.NLRP3 inflammasome inhibitor and gasdermin D(GSDMD)inhibitor were used to explore the mechanism of Prdx1-induced pyroptosis.RESULTS The mRNA and protein levels of Prdx1 were significantly increased in the tumor tissues of patients with CRC.rPrdx1 inhibited the viability,proliferation,migration and invasion of RKO and SW480 colon cancer cells.Further study found that rPrdx1 inhibited the malignant biological behaviors of CRC cells by inducing pyroptosis rather than apoptosis and necroptosis.Mechanistically,rPrdx1 induces pyroptosis of CRC cells by activating the NLRP3 inflammasome/GSDMD pathway.CONCLUSION Prdx1 induces pyroptosis by activating the NLRP3 inflammasome/GSDMD pathway,thereby inhibiting the malignant biological behavior of RKO and SW480 colon cancer cells.展开更多
Inflammatory bowel disease(IBD),including ulcerative colitis and Crohn’s disease,is a chronic intestinal inflammation with complex pathogenesis.Pyroptosis a pro-inflammatory programmed cell death mediated by gasdermi...Inflammatory bowel disease(IBD),including ulcerative colitis and Crohn’s disease,is a chronic intestinal inflammation with complex pathogenesis.Pyroptosis a pro-inflammatory programmed cell death mediated by gasdermin D(GSDMD)cleavage plays a pivotal role in disease progression through nucleotide-binding oligomerization domain-like receptor family pyrin domain-containing 3(NLRP-3)/caspase-1 classical and caspase-4/5/11 non-classical pathways.Targeting pyroptosis has emerged as a promising therapeutic strategy,with recent advances highlighting the potential of pyroptosis inhibitors such as small-molecule compounds,biologics,and repurposed drugs that specifically target NLRP3,caspases,or GSDMD to suppress inflammasome activation,block pore formation,and mitigate downstream inflammation.This review systematically summarizes the mechanisms and therapeutic effects of these inhibitors,while addressing critical challenges including drug specificity,delivery efficiency,and long-term safety,and explores their potential in combination therapies with existing IBD treatments to enhance clinical efficacy.By integrating preclinical and clinical evidence,we provide valuable insights into the translational prospects of pyroptosis-targeted therapies for precision management of IBD.展开更多
基金supported by grants from the National Key Research and Development Program of China(2022YFD2401001)Central Public-interest Scientific Institution Basal Research Fund,CAFS(NO.2023TD81 and NO.20603022023018)China Agriculture Research System(CARS47-G19).
文摘Hypoxia is one of the major environmental stressors,frequently resulting in serious yield losses for maricultured large yellow croaker(Larimichthys crocea).We know that pyroptosis contributes to tissue damage under hypoxic conditions.However,whether GSDMEs-mediated pyroptosis is involved in hypoxia-induced tissue damage in fish remains unclear.In the present study,two Lcgsdme genes,Lcgsdmea/b,were cloned and characterized from the large yellow croaker.Both genes contain a conserved N-terminalpore-forming domain,a C-terminal auto-inhibitory domain,and a flexible hinge region.After hypoxia stress,the expression of Lcgsdmea/b transcripts and proteins in the liver were significantly higher than in unstressed fish.The proteins of LcGSDMEa/b could be cleaved under hypoxic conditions.Compared to LcGSDMEb,the expression of LeGSDMEa was higher in both mRNA and protein levels,thus exhibiting a stronger response during hypoxia stress.Furthermore,after 48 h of hypoxia stress,approximately 65%liver cells exhibited abnormalities,with pyroptosis being detected using a transmission electron microscope.TUNEI/LeGSDMEa double staining assay revealed a high expression of LcGSDMEa in the dead cells.We observed a significant up-regulation of pyroptosis pathway genes(asc,caspase-3)and pro-inflammatory cytokine genes(il-1β,il-18).After simultaneous knockdown of Legsdmea/b in vivo,the liver exhibited better health compared to the control group,with less cell swelling and vacuolation.Taken together,these findings demonstrate that hypoxia stress could activate LcGSDMEa/b and induce pyroptosis in the liver of large yellow croakers,thereby contributing to tissue damage.Our study improves the understanding of hypoxia-induced tissue damage in fish,and provides new clues for protecting fish against hypoxia-induced damage.
基金supported by grants from the National Key Research and Development Program of China,No.2017YFA0105400(to LR)the Key Research and Development Program of Guangdong Province,No.2019B020236002(to LR)the National Natural Science Foundation of China,Nos.81972111(to LZ),81772349(to BL).
文摘Microglia,the resident monocyte of the central nervous system,play a crucial role in the response to spinal cord injury.However,the precise mechanism remains unclear.To investigate the molecular mechanisms by which microglia regulate the neuroinflammatory response to spinal cord injury,we performed single-cell RNA sequencing dataset analysis,focusing on changes in microglial subpopulations.We found that the MG1 subpopulation emerged in the acute/subacute phase of spinal cord injury and expressed genes related to cell pyroptosis,sphingomyelin metabolism,and neuroinflammation at high levels.Subsequently,we established a mouse model of contusive injury and performed intrathecal injection of siRNA and molecular inhibitors to validate the role of ceramide synthase 5 in the neuroinflammatory responses and pyroptosis after spinal cord injury.Finally,we established a PC12-BV2 cell co-culture system and found that ceramide synthase 5 and pyroptosis-associated proteins were highly expressed to induce the apoptosis of neuron cells.Inhibiting ceramide synthase 5 expression in a mouse model of spinal cord injury effectively reduced pyroptosis.Furthermore,ceramide synthase 5-induced pyroptosis was dependent on activation of the NLRP3 signaling pathway.Inhibiting ceramide synthase 5 expression in microglia in vivo reduced neuronal apoptosis and promoted recovery of neurological function.Pla2g7 formed a“bridge”between sphingolipid metabolism and ceramide synthase 5-mediated cell death by inhibiting the NLRP3 signaling pathway.Collectively,these findings suggest that inhibiting ceramide synthase 5 expression in microglia after spinal cord injury effectively suppressed microglial pyroptosis mediated by NLRP3,thereby exerting neuroprotective effects.
基金supported by grants from the National Natural Science Foundation of China(8170060495 and 82170682)。
文摘Background:Cisplatin triggers Gasdermin E(GSDME)cleavage,causing membrane bubble formation,content release,and inflammation.Caspase-3 activation initiates GSDME cleavage,and thus inhibiting this pathway mitigates cisplatin-induced pyroptosis in hepatocytes.This study aimed to delve into how cisplatin induces liver injury via pyroptosis.Methods:For animal experiments,C57BL/6J mice were divided into three groups:control,liver injury model group,and Ac-DMLD-CMK(caspase-3 inhibitor)intervention group.The liver histology was evaluated by hematoxylin and eosin staining,immunohistochemistry,immunofluorescence and TUNEL staining.The mRNA and protein levels were detected by real-time polymerase chain reaction(PCR)and Western blot analysis.For in vitro experiments,HL-7702 cells were treated with cisplatin or GSDME siRNA.Cell pyroptosis was determined via cellular morphology,cytotoxicity and viability detection,flow cytometric assay,and Western blot detection for the expression of pyroptosis-related proteins.Results:Cisplatin-induced distinct liver morphological changes,hepatocellular injury,and inflammation in mice,along with elevated serum alanine aminotransferase(ALT)and aspartate aminotransferase(AST)levels and increased pro-inflammatory cytokine expression.Heightened macrophage infiltration and hepatocellular death indicated cisplatin-induced hepatotoxicity.Cisplatin upregulated GSDME activation,along with Bax-mediated caspase-3 cleavage both in vivo and in vitro,implicating caspase-3/GSDME-dependent pyroptosis in liver injury.Treatment with Ac-DMLD-CMK ameliorated cisplatin-induced liver injury,reducing hepatocellular lesions,serum ALT and AST levels,cytokine expression,macrophage infiltration,and hepatocyte death.Ac-DMLD-CMK also attenuated GSDME-dependent pyroptosis post-cisplatin induction,as evidenced by decreased GSDME expression,Bax upregulation,and cleaved caspase-3 activation.For HL-7702 cells,GSDME siRNA transfection reduced GSDME expression,attenuated typical signs of cisplatin-induced pyroptosis,partially restored cell viability,and significantly inhibited cytotoxicity and a decrease in the proportion of propidium iodide-positive cells,indicating protection against cisplatininduced hepatocyte pyroptosis.Conclusions:Our study underscores the role of the caspase-3/GSDME signaling pathway in mediating cisplatin-induced hepatotoxicity,particularly in cases of excessive or cumulative cisplatin exposure.These findings suggest that targeting GSDME could represent a promising therapeutic approach to mitigate cisplatin-induced liver damage.
基金Supported by the National Natural Science Foundation of China,No.82302454Natural Science Foundation of Hunan Province,No.2025JJ60797,No.2025JJ60734 and No.2022JJ40718Guangdong Basic and Applied Basic Research Foundation,No.2022A1515111063.
文摘BACKGROUND Damage associated molecular patterns(DAMPs)are vital for the immunogenic cell death of cancer cells and can enhance the anti-tumor activity of immune cells in colorectal cancer(CRC).Peroxiredoxin 1(Prdx1),an important DAMP,is highly expressed in various tumor tissues including CRC.However,the role of Prdx1 in CRC remains unknown.AIM To investigate the effect and mechanisms of Prdx1 on CRC.METHODS Patients diagnosed with CRC in our medical center were included in this study to verify the expression of Prdx1 in cancer tissues.Recombinant Prdx1(rPrdx1)was used to stimulate RKO and SW480 colon cancer cells.The cell survival rate,migration,proliferation and invasion ability were assessed.Transmission electron microscopy,TUNEL assay,lactate dehydrogenase release assay,and Western blot were used to determine the effect of Prdx1 on pyroptosis.NLRP3 inflammasome inhibitor and gasdermin D(GSDMD)inhibitor were used to explore the mechanism of Prdx1-induced pyroptosis.RESULTS The mRNA and protein levels of Prdx1 were significantly increased in the tumor tissues of patients with CRC.rPrdx1 inhibited the viability,proliferation,migration and invasion of RKO and SW480 colon cancer cells.Further study found that rPrdx1 inhibited the malignant biological behaviors of CRC cells by inducing pyroptosis rather than apoptosis and necroptosis.Mechanistically,rPrdx1 induces pyroptosis of CRC cells by activating the NLRP3 inflammasome/GSDMD pathway.CONCLUSION Prdx1 induces pyroptosis by activating the NLRP3 inflammasome/GSDMD pathway,thereby inhibiting the malignant biological behavior of RKO and SW480 colon cancer cells.
基金Supported by the Science and Technology Program of Gansu Province,No.23JRRA1015.
文摘Inflammatory bowel disease(IBD),including ulcerative colitis and Crohn’s disease,is a chronic intestinal inflammation with complex pathogenesis.Pyroptosis a pro-inflammatory programmed cell death mediated by gasdermin D(GSDMD)cleavage plays a pivotal role in disease progression through nucleotide-binding oligomerization domain-like receptor family pyrin domain-containing 3(NLRP-3)/caspase-1 classical and caspase-4/5/11 non-classical pathways.Targeting pyroptosis has emerged as a promising therapeutic strategy,with recent advances highlighting the potential of pyroptosis inhibitors such as small-molecule compounds,biologics,and repurposed drugs that specifically target NLRP3,caspases,or GSDMD to suppress inflammasome activation,block pore formation,and mitigate downstream inflammation.This review systematically summarizes the mechanisms and therapeutic effects of these inhibitors,while addressing critical challenges including drug specificity,delivery efficiency,and long-term safety,and explores their potential in combination therapies with existing IBD treatments to enhance clinical efficacy.By integrating preclinical and clinical evidence,we provide valuable insights into the translational prospects of pyroptosis-targeted therapies for precision management of IBD.
