Background:Intestinal ischemia/reperfusion(II/R)is a severe condition with high mortality and limited treatment options.Extracellular vesicles that are derived from bone marrow mesenchymal stem cells(BM-MSC-EVs)exhibi...Background:Intestinal ischemia/reperfusion(II/R)is a severe condition with high mortality and limited treatment options.Extracellular vesicles that are derived from bone marrow mesenchymal stem cells(BM-MSC-EVs)exhibit therapeutic potential in alleviating II/R injury.However,the mechanism by which BM-MSC-EVs fulfill this function requires further characterization.The ubiquitin-proteasome system plays an essential role in II/R,but the functions of individual ubiquitination regulators such as ubiquitin-specific proteases(USPs)in this process remain incompletely understood.Methods:An II/R cellular model was established by using IEC-6 intestinal epithelial cells with oxygen-glucose deprivation/reperfusion(OGD/R)treatment.The expression of USPs was evaluated by using quantitative polymerase chain reaction and Western blot.The role of USP38 on the viability,apoptosis,migration,and reactive oxygen species(ROS)levels in OGD/R-treated IEC-6 cells were measured by using CCK-8,Annexin V/PI staining,transwell assay,and 20,70-dichlorofluorescin diacetate(DCFDA)staining,respectively.The interaction between USP38 and BIRC5 was explored by using co-immunoprecipitation(Co-IP)and the ubiquitination level and stability of BIRC5 were examined by using Western blot.USP38-overexpressing BM-MSC-EVs were produced to treat OGD/R-treated IEC-6 cells.Results:USP38 expression was significantly downregulated in OGD/R-treated IEC-6 cells.Incubation of these cells with BM-MSC-EVs substantially elevated the USP38 expression,resulting in improved viability,reduced apoptosis,enhanced migration,and decreased ROS levels.Furthermore,overexpression of USP38 in BM-MSC-EVs further enhanced their protective effect on OGD/R-treated IEC-6 cells.At the molecular level,USP38 interacts with and stabilizes BIRC5 by decreasing its ubiquitination.Knock-down of BIRC5 abolished the protective effect of excessive USP38 on OGD/R-treated IEC-6 cells.Conclusion:USP38 protects intestinal epithelial cells from I/R injury by enhancing the stability of BIRC5.展开更多
基金supported by Natural Science Foundation of Fujian Province,China[2021J011357]。
文摘Background:Intestinal ischemia/reperfusion(II/R)is a severe condition with high mortality and limited treatment options.Extracellular vesicles that are derived from bone marrow mesenchymal stem cells(BM-MSC-EVs)exhibit therapeutic potential in alleviating II/R injury.However,the mechanism by which BM-MSC-EVs fulfill this function requires further characterization.The ubiquitin-proteasome system plays an essential role in II/R,but the functions of individual ubiquitination regulators such as ubiquitin-specific proteases(USPs)in this process remain incompletely understood.Methods:An II/R cellular model was established by using IEC-6 intestinal epithelial cells with oxygen-glucose deprivation/reperfusion(OGD/R)treatment.The expression of USPs was evaluated by using quantitative polymerase chain reaction and Western blot.The role of USP38 on the viability,apoptosis,migration,and reactive oxygen species(ROS)levels in OGD/R-treated IEC-6 cells were measured by using CCK-8,Annexin V/PI staining,transwell assay,and 20,70-dichlorofluorescin diacetate(DCFDA)staining,respectively.The interaction between USP38 and BIRC5 was explored by using co-immunoprecipitation(Co-IP)and the ubiquitination level and stability of BIRC5 were examined by using Western blot.USP38-overexpressing BM-MSC-EVs were produced to treat OGD/R-treated IEC-6 cells.Results:USP38 expression was significantly downregulated in OGD/R-treated IEC-6 cells.Incubation of these cells with BM-MSC-EVs substantially elevated the USP38 expression,resulting in improved viability,reduced apoptosis,enhanced migration,and decreased ROS levels.Furthermore,overexpression of USP38 in BM-MSC-EVs further enhanced their protective effect on OGD/R-treated IEC-6 cells.At the molecular level,USP38 interacts with and stabilizes BIRC5 by decreasing its ubiquitination.Knock-down of BIRC5 abolished the protective effect of excessive USP38 on OGD/R-treated IEC-6 cells.Conclusion:USP38 protects intestinal epithelial cells from I/R injury by enhancing the stability of BIRC5.
