摘要
目的探讨左西孟旦通过PTEN/Akt通路抑制脂多糖(LPS)引起C2C12细胞凋亡的机制。方法CCK-8法检测不同浓度左西孟旦对C2C12细胞存活率的影响,选择有保护作用的左西孟旦浓度。将C2C12细胞随机分为4组:空白对照组(CON组)、左西孟旦预处理后的对照组(CON+L组)、LPS处理组(LPS组)、左西孟旦预处理后的LPS处理组(LPS+L组)。采用CCK-8法检测C2C12细胞的存活率;hoechst33342染色细胞核观察细胞的凋亡情况;采用实时PCR、Western blotting检测C2C12细胞PTEN/Akt通路凋亡指标的mRNA及蛋白表达水平。siRNA-PTEN转染C2C12细胞,敲除PTEN基因,检测其对凋亡通路蛋白表达的影响。结果左西孟旦能够提高LPS损伤后C2C12细胞的存活率,降低凋亡率。siRNA-PTEN抑制了PTEN基因的表达,PTEN/Akt信号通路相关mRNA与蛋白发生相应改变,导致C2C12细胞凋亡率下降。结论左西孟旦可以通过PTEN/Akt通路抑制LPS引起的C2C12细胞凋亡。
Objective To investigate whether levosimendan can inhibit the apoptosis of C2C12 cells induced by lipopolysaccharide(LPS)through the PTEN/Akt pathway.Methods C2C12 cells were randomly divided into four groups:blank control group,control group comprising cells treated with levosimendan only,LPS-treated group,and a group comprising cells pretreated with levosimendan for 24 h a subsequently treated with LPS for 48 h.The survival rate of C2C12 cells was determined via the CCK-8 method,and cell apoptosis was assessed via Hoechst 33342 staining.The mRNA and protein expression levels of PTEN/Akt pathway components were evaluated via RT-qPCR and Western blotting,respectively.C2C12 cells were also transfected with siRNA to knockdown the PTEN gene,and the effect on the protein expression of apoptotic pathway components was determined.Results Levosimendan increased the survival rate and decreased the apoptosis rate of C2C12 cells after LPS treatment.PTEN gene expression was inhibited by siRNA and the mRNA and protein levels of PTEN/Akt signaling pathway components changed correspondingly.Furthermore,the apoptosis rate of C2C12 cells decreased.Conclusion Levosimendan can inhibit LPS-induced C2C12 cell apoptosis via the PTEN/Akt pathway.
作者
张苗苗
刘雨佳
张甦
焦光宇
ZHANG Miaomiao;LIU Yujia;ZHANG Su;JIAO Guangyu(Department of Pulmonary and Critical Care Medicine,Shengjing Hospital of China Medical University,Shenyang 110004,China;Department of Chinese Traditional Medicine,Liaoning University of Traditional Chinese Medicine,Shenyang 110032,China)
出处
《中国医科大学学报》
北大核心
2024年第2期121-126,共6页
Journal of China Medical University
基金
辽宁省应用基础研究计划联合计划项目(2022JH2/101500051)。