芪骨胶囊含药血清对地塞米松诱导C2C12细胞萎缩的保护作用  

Protective effects of Qigu Capsule-medicated serum on C2C12 cells atrophy induced by dexamethasone

作　　者：石金玉 杨宗睿 葛海雅 郭海玲[1,2] 詹红生[1,2] SHI Jin-yu;YANG Zong-rui;GE Hai-ya;GUO Hai-ling;ZHAN Hong-sheng(Shi s Center of Orthopedics and Traumatology,Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine,Shanghai 200120,China;Institute of Traumatology&Orthopedics,Shanghai Academy of Traditional Chinese Medicine,Shanghai 200120,China)

机构地区：[1]上海中医药大学附属曙光医院石氏伤科医学中心,上海200120 [2]上海市中医药研究院骨伤科研究所,上海200120

出　　处：《中成药》2023年第12期3914-3921,共8页Chinese Traditional Patent Medicine

基　　金：国家中医药管理局项目(2019);上海市科学技术委员会项目(20MC1920600);上海市卫生健康委员会项目(shslczdzk03901);上海市中医药发展办公室项目[ZY(2018-2020)-CCCX-1010];上海市申康医院发展中心项目(SHDC2020CR3090B);上海中医药大学项目(2018)。

摘　　要：目的观察芪骨胶囊含药血清对地塞米松诱导的C2C12肌管细胞萎缩的保护作用。方法制备芪骨胶囊含药血清;体外培养C2C12细胞,分为正常对照组、地塞米松组、10%空白血清组、10%芪骨胶囊含药血清组,给予相应药物处理48 h后,除正常对照组外再给予10μmol/L地塞米松继续培养24 h,CCK-8法检测各组细胞活力。用2%马血清诱导C2C12细胞分化6~7 d,按上述分组干预肌管细胞,测量各组肌管细胞直径,Western blot法检测MyHC、MuRF-1、Atrogin-1及PI3K/Akt信号通路相关蛋白表达。加入PI3K抑制剂LY294002,测量肌管细胞直径,检测MuRF-1、Atorgin-1、PI3K/Akt信号通路相关蛋白表达。结果与正常对照组比较,地塞米松组及空白血清组C2C12细胞活力降低(P<0.01),肌管细胞直径减少(P<0.01),MuRF-1、Atrogin-1蛋白表达升高(P<0.01),MyHC蛋白表达及PI3K、Akt、mTOR、FoxO3a磷酸化水平降低(P<0.01);与地塞米松组比较,芪骨胶囊含药血清组细胞活力升高(P<0.01),肌管细胞直径增加(P<0.01),MuRF-1、Atrogin-1蛋白表达降低(P<0.01),MyHC蛋白表达及PI3K、Akt、mTOR、FoxO3a的磷酸化水平升高(P<0.05,P<0.01)。芪骨胶囊含药血清联合LY294002干预后,芪骨胶囊含药血清的抗肌管萎缩效应消失(P<0.01),MuRF-1、Atorgin-1蛋白表达升高(P<0.01),而PI3K、Akt、mTOR、FoxO3a磷酸化水平降低(P<0.05,P<0.01)。结论芪骨胶囊含药血清能减轻地塞米松诱导的C2C12肌管细胞萎缩,其机制可能与激活PI3K/Akt信号通路相关。AIM To observe the protective effects of Qigu Capsule-medicated serum on C2C12 myotube cell atrophy induced by dexamethasone.METHODS The Qigu Capsule-medicated serum was prepared.C2C12 cells cultured in vitro were divided into the normal control group,the dexamethasone group,the 10%blank serum group and the 10%Qigu Capsule-medicated serum group for 48 h,corresponding drug treatment,followed by 24 h culture with 10μmol/L dexamethasone except the control group,and had cell viability detection by CCK-8 method.With their myotube cells intervened accordingly by the aforementioned regime following 6-7 days differentiation with 2%horse serum induction,the C2C12 cells had their myotube cells diameter measured;and the expressions of proteins related to MyHC,MuRF-1,Atrogin-1 and PI3K/Akt signaling pathway detected by Western blot.The impact of PI3K inhibitor LY294002 on the diameter of myotube cells and the expression of MuRF-1,Atorgin-1 and PI3K/Akt signaling pathway related proteins were detected as well.RESULTS Compared with the normal control group,the dexamethasone group and the blank serum group were observed with decreased viability of C2C12 cells(P<0.01),decreased diameter of myotube cells(P<0.01),increased protein expressions of MuRF-1 and Atrogin-1(P<0.01),and decreased expression of MyHC protein and phosphorylation levels of PI3K,Akt,mTOR and FoxO3a(P<0.01).Compared with the dexamethasone group,the Qigu Capsule-medicated serum group showed increased viability of C2C12 cells(P<0.01);increased diameter of myotube cells(P<0.01);decreased protein expressions of MuRF-1 and Atrogin-1(P<0.01);and increased expression of MyHC protein and phosphorylation levels of PI3K,Akt,mTOR and FoxO3a(P<0.05,P<0.01).The intervention of LY294002 upon the Qigu Capsule-medicated serum group offset the anti-muscular tube atrophy effect(P<0.01),increased the protein expressions of MuRF-1 and Atorgin-1(P<0.01),and decreased the phosphorylation levels of PI3K,Akt,mTOR and FoxO3a(P<0.05,P<0.01).CONCLUSION The Qigu Capsule-medicated serum ca

关 键 词：芪骨胶囊 C2C12成肌细胞 细胞萎缩 PI3K/AKT信号通路 

分 类 号：R285.5[医药卫生—中药学]