Background:Sepsis,a type of systemic disease,can impact nearly all organs,tissues and cells.Among them,endothelial cells are amongst thefirst to be affected and respond to the insult.In this study,we investigated the p...Background:Sepsis,a type of systemic disease,can impact nearly all organs,tissues and cells.Among them,endothelial cells are amongst thefirst to be affected and respond to the insult.In this study,we investigated the protective effects of microRNA-30a-5p(miR-30a-5p)on human pulmonary microvascular endothelial cells(HPMECs)treated with lipolysaccharide(LPS).Methods:An in vitro model of sepsis was established in HPMECs with the use of LPS.Transfecting with different tools(mimetic and inhibitor)to modify miR-30a-5p expression.Cell viability,proliferation and apoptosis were detected by the CCK-8 assay,the EdU kit andfluorescence staining,respectively.The autophagy-related protein and mRNA expression,the number of autophagosomes were separately examined through Western blot analysis,qPT-PCR and confocal microscopy.TargetScan and the luciferase reporter assays were used to probe target genes interacting with miR-30a-5p.Results:LPS caused a reduction in the viability and proliferation of HPMECs,as well as an elevation in the number of apoptotic cells.Subsequently,we observed that miR-30a-5p might play a role in preventing LPS-induced inhibition of cell damage and decreasing HPMEC apoptosis,suggesting the potential function of miR-30a-5p in this injury process.Finally,we confirmed that miR-30a-5p exerts its protective effect by regulating cell autophagy,possibly by targeting Beclin-1.Conclusion:Our study provided evidence that autophagy is a crucial aspect in the protective role of miR-30a-5p against LPS-induced HPMEC injury,identifying a promising target for sepsis-related endothelial cell injury.展开更多
基金This study is supported in part by Zhejiang Provincial Natural Science Foundation of China(Grant No.LTGD23H150001)the Medical and Health Research Project of Zhejiang Province(Grant No.2023KY212)the High-Level Talent Research Project of Hangzhou Vocational&Technical College(Grant No.HZYGCC202229).
文摘Background:Sepsis,a type of systemic disease,can impact nearly all organs,tissues and cells.Among them,endothelial cells are amongst thefirst to be affected and respond to the insult.In this study,we investigated the protective effects of microRNA-30a-5p(miR-30a-5p)on human pulmonary microvascular endothelial cells(HPMECs)treated with lipolysaccharide(LPS).Methods:An in vitro model of sepsis was established in HPMECs with the use of LPS.Transfecting with different tools(mimetic and inhibitor)to modify miR-30a-5p expression.Cell viability,proliferation and apoptosis were detected by the CCK-8 assay,the EdU kit andfluorescence staining,respectively.The autophagy-related protein and mRNA expression,the number of autophagosomes were separately examined through Western blot analysis,qPT-PCR and confocal microscopy.TargetScan and the luciferase reporter assays were used to probe target genes interacting with miR-30a-5p.Results:LPS caused a reduction in the viability and proliferation of HPMECs,as well as an elevation in the number of apoptotic cells.Subsequently,we observed that miR-30a-5p might play a role in preventing LPS-induced inhibition of cell damage and decreasing HPMEC apoptosis,suggesting the potential function of miR-30a-5p in this injury process.Finally,we confirmed that miR-30a-5p exerts its protective effect by regulating cell autophagy,possibly by targeting Beclin-1.Conclusion:Our study provided evidence that autophagy is a crucial aspect in the protective role of miR-30a-5p against LPS-induced HPMEC injury,identifying a promising target for sepsis-related endothelial cell injury.
