补肺健脾方对香烟烟雾提取物诱导大鼠成肌细胞AKT/FOXO3a/Bim信号通路的影响

Effects of Bufei Jianpi Decoction on AKT/FOXO3a/Bim signaling pathway in L6 cell induced by cigarette smoke extract

目的:探讨补肺健脾方对香烟烟雾提取物(CSE)诱导大鼠成肌细胞(L6细胞)蛋白激酶B(AKT)/叉头转录因子O亚型3a(FOXO3a)/Bcl-2相互作用的死亡介质(Bim)信号通路的影响。方法:正常培养的L6细胞随机分为正常对照组、CSE组、补肺健脾方(FP)组,分别给予10%空白血清、10%空白血清+10%CSE、10%补肺健脾方血清+10%CSE培养基培养24h后,采用荧光定量PCR技术检测AKT、FOXO3a、Bim mRNA表达的变化;采用Western Blot技术检测AKT、p-AKT、FOXO3a、p-FOXO3a和Bim蛋白表达的变化。结果:与正常对照组比较,CSE组L6细胞FOXO3a和Bim mRNA及蛋白表达显著升高(P<0.01,P<0.05),p-AKT、p-FOXO3a蛋白表达显著下降(P<0.05);与CSE组比较,FP组FOXO3a mRNA表达显著下降(P<0.05),p-AKT、p-FOXO3a蛋白表达显著增加(P<0.05),Bim mRNA和蛋白表达均显著降低(P<0.05)。结论:补肺健脾方能够调控L6细胞AKT/FOXO3a/Bim信号通路,从而减少骨骼肌细胞凋亡,改善骨骼肌功能障碍。

Objective:To observe the effects of Bufei Jianpi Decoction on cigarette smoke extract(CSE)induced myoblast(L6)protein kinase B(AKT)/forkhead box O3a(FOXO3a)/Bcl-2 interacting mediator of cell death(Bim)signaling pathway.Methods:Normally cultured L6 cells were randomly divided into control group,cigarette smoke extract(CSE)group,and Bufei Jianpi Decoction(FP)group.The L6 cells were treated with 10%control serum(control group),10%control serum+10%CSE(CSE group),10%Bufei Jianpi granules serum+10%CSE(FP group).The expressions of AKT,FOXO3a,and Bim mRNA were examined by Real-time fluorescence quantitative polymerase chain reaction(qPCR);Western Blot was used to detect changes in AKT,p-AKT,FOXO3a,p-FOXO3a and Bim protein expression.Results:In CSE group,FOXO3a and Bim mRNA and protein were higher,while p-AKT and p-FOXO3a protein were markedly lower than in control group(P<0.01,P<0.05).Compared with CSE group,p-AKT and p-FOXO3a protein were significantly improved(P<0.05),while Bim mRNA and protein were observably lower in FP group(P<0.05).Conclusion:Bufei Jianpi Decoction can decrease cell apoptosis and improve skeletal muscle dysfunction by regulatting AKT/FOXO3a/Bim signaling pathway.