槲皮素调控AKT1/AMPK/Foxo3a信号通路抑制C2C12细胞凋亡  

Quercetin Regulates the AKT1/AMPK/Foxo3a Signaling Pathway to Inhibit Apoptosis in C2C12 Cells

作　　者：张祥生 牛晓莹 刘源 叶茂林 柴爽 马江涛 ZHANG Xiangsheng;NIU Xiaoying;LIU Yuan;YE Maolin;CHAI Shuang;MA Jiangtao(Luoyang Orthopedic-Traumatological Hospital of Henan Province(Henan Provincial Orthopedic Hospital),Zhengzhou 450046,China)

机构地区：[1]河南省洛阳正骨医院(河南省骨科医院),河南郑州450046

出　　处：《中医药信息》2024年第11期15-22,共8页Information on Traditional Chinese Medicine

基　　金：河南省自然科学基金项目(222300420198);河南省洛阳正骨医院博士科研启动项目(2023BS03,2024BS04)。

摘　　要：目的:基于AKT1/AMPK/Foxo3a信号通路探讨槲皮素对C2C12细胞增殖、成肌分化和凋亡的影响。方法:体外培养C2C12细胞进行成肌诱导,分别采用CCK8法(Cell Counting Kit-8)、ATP染色法及TUNEL法检测槲皮素对C2C12细胞增殖活力、成肌能力和凋亡的影响;实时荧光定量PCR(qRT-PCR)和蛋白免疫印迹(Western blot)分别检测MyoD1和Myogenin基因和蛋白表达,筛选合适浓度。再采用合适浓度的槲皮素或AKT1通路抑制剂(LY294002)干预成肌诱导后的C2C12细胞,ATP法和TUNEL法分别检测C2C12细胞成肌能力和凋亡水平,Western blot检测p-AKT1、p-AMPK和p-Foxo3a蛋白表达水平。结果:①在一定范围内,槲皮素呈浓度依赖性促进C2C12增殖,12.5μmol/L的槲皮素显著促进C2C12细胞成肌分化,能显著提高成肌标志物MyoD1和Myogenin基因和蛋白表达,因此本研究选择12.5μmol/L的槲皮素用于后续研究。②10μmol/L LY294002显著抑制C2C12细胞成肌分化,促进细胞凋亡,抑制p-AKT1和p-AMPK蛋白表达,促进p-Foxo3a蛋白表达;12.5μmol/L槲皮素联合10μmol/L LY294002显著促进C2C12细胞成肌分化,抑制细胞凋亡,促进p-AKT1和p-AMPK蛋白表达,抑制p-Foxo3a蛋白表达。结论:槲皮素通过调控AKT1/AMPK/Foxo3a信号通路的磷酸化促进C2C12细胞增殖和分化,抑制细胞凋亡。Objective:To explore the effects of quercetin on proliferation,myogenic differentiation,and apoptosis of C2C12 cells through the AKT1/AMPK/Foxo3a signaling pathway.Methods:C2C12 cells were cultured in vitro and induced for myogenic differentiation.Cell Counting Kit-8(CCK-8),ATP assay,and TUNEL assay were used to evaluate the effects of quercetin on cell proliferation,myogenic capacity,and apoptosis.Real-time quantitative PCR(qRT-PCR)and Western blot were employed to assess the expression of MyoD1 and Myogenin genes and proteins to determine the appropriate concentration.Subsequently,C2C12 cells induced for myogenesis were treated with quercetin at the appropriate concentration or AKT1 pathway inhibitor(LY294002).ATP assay and TUNEL assay were used to evaluate myogenic capacity and apoptosis levels,respectively.Western blot was performed to measure the expression levels of p-AKT1,p-AMPK,and p-Foxo3a proteins.Results:(1)Quercetin promoted C2C12 proliferation in a concentration-dependent manner within a certain range.At 12.5μmol/L,quercetin significantly enhanced myogenic differentiation of C2C12 cells and upregulated the expression of myogenic markers MyoD1 and Myogenin genes and proteins.Therefore,12.5μmol/L quercetin was selected for subsequent studies.(2)LY294002 at 10μmol/L significantly inhibited myogenic differentiation of C2C12 cells,promoted apoptosis,inhibited the expression of p-AKT1 and p-AMPK proteins,and promoted p-Foxo3a protein expression.Combination treatment with 12.5μmol/L quercetin and 10μmol/L LY294002 significantly promoted myogenic differentiation,inhibited apoptosis of C2C12 cells,promoted the expression of p-AKT1 and p-AMPK proteins,and inhibited p-Foxo3a protein expression.Conclusion:Quercetin promotes proliferation and differentiation of C2C12 cells and inhibits apoptosis by regulating the phosphorylation of the AKT1/AMPK/Foxo3a signaling pathway.

关 键 词：槲皮素 AKT1/AMPK/Foxo3a信号通路 C2C12细胞 成肌分化 凋亡 

分 类 号：R285[医药卫生—中药学]