藤茶总黄酮调节C2C12细胞胰岛素抵抗的作用和机制研究  

Study on Effects and Mechanisms of Regulation of Total Flavonoids from Ampelopsis Grossedentata on Insulin Resistance of C2C12 Cells

作　　者：李可 吴丽丽[3] 秦灵灵 胡玉立 吴悠 张程斐[2] 张秀媛 刘铜华[2,3] LI Ke;WU Lili;QIN Lingling;HU Yuli;WU You;ZHANG Chengfei;ZHANG Xiuyuan;LIU Tonghua(Shunyi Hospital,Beijing Traditional Chinese Medicine Hospital,Beijing 101300,China;Dongfang Hospital Affiliated to Beijing University of Chinese Medicine,Beijing 100078,China;Key Laboratory of Chinese Medicine Health-cultivation of The Ministry of Education,Beijing University of Chinese Medicine,Beijing 100029,China;Department of Science & Technology,Beijing University of Chinese Medicine,Beijing 100029,China)

机构地区：[1]北京中医医院顺义医院,北京101300 [2]北京中医药大学东方医院,北京100078 [3]北京中医药大学教育部中医养生学重点实验室,北京100029 [4]北京中医药大学科技处,北京100029

出　　处：《世界中医药》2021年第19期2885-2890,共6页World Chinese Medicine

基　　金：国家自然科学基金青年科学基金项目(82004338);中医药防治糖尿病及其并发症学科创新引智基地项目(B20055)

摘　　要：目的:观察藤茶总黄酮对棕榈酸诱导的C2C12肌管细胞胰岛素抵抗模型葡萄糖利用的影响,并从IRS-1/PI3K p85/AKT/GLUT4信号转导通路探讨其作用机制。方法:用含2%马血清的高糖培养基(DMEM)培养小鼠C2C12成肌细胞,诱导为成熟的肌管细胞后,用0.5 mmol/L棕榈酸诱导培养16 h建立细胞胰岛素抵抗模型。用CCK-8法测定不同浓度的藤茶总黄酮(5μg/mL、10μg/mL、20μg/mL、30μg/mL、50μg/mL、80μg/mL)对C2C12细胞活性的影响,并确定后续实验的藤茶总黄酮高、中、低剂量。将C2C12细胞诱导成熟的肌管细胞分为正常组(NC)、高糖模型组(DM)和藤茶总黄酮高、中、低剂量组(AGH、AGM、AGL),检测各组细胞的葡萄糖消耗量,用Western Blotting法检测藤茶总黄酮对IRS-1/PI3K p85/AKT/GLUT4信号转导通的蛋白表达的影响。结果:藤茶总黄酮浓度在50μg/mL时,细胞成活率最高。据C2C12细胞活性检测,得到用于后续细胞实验的藤茶总黄酮高、中、低浓度分别为80μg/mL、50μg/mL、30μg/mL。藤茶总黄酮高、中、低剂量组葡萄糖消耗量与高糖模型组比较显著增加(P<0.01)。藤茶总黄酮高、中剂量组显著提升IRS-1/PI3K p85/AKT/GLUT4信号通路的蛋白表达(P<0.01)。结论:藤茶总黄酮可改善C2C12细胞葡萄糖消耗量从而改善胰岛素抵抗,其作用机制可能通过调节IRS-1/PI3K p85/AKT/GLUT4信号通路实现。Objective:To observe the effects of total flavonoids from Ampelopsis grossedentata on glucose utilization in insulin resistance models of C2C12 myotube cells induced by palmitic acid,and to explore its mechanism from IRS-1/PI3K p85/AKT/GLUT4 signaling transduction pathways.Methods:Mouse C2C12 myoblasts were cultured in high glucose medium(DMEM)containing 2%horse serum,and then induced into mature myotube cells,which were cultured for 16 hours with 0.5mmol/L palmitic acid to establish the cellular insulin resistance models.The CCK-8 method was used to determine the effects of the total flavonoids from Ampelopsis grossedentata at different concentrations(5μg/mL,10μg/mL,20μg/mL,30μg/mL,50μg/mL,80μg/mL)on the activity of C2C12 cells,and determine the high,medium and low doses of total flavonoids from Ampelopsis grossedentata in subsequent experiments.The myotube cells induced by C2C12 cells were divided into normal group(NC),high glucose model group(DM)and total flavonoids from Ampelopsis grossedentata in high,medium,and low dose groups(AGH,AGM,AGL).The glucose consumption of each group cells was measured.Western Blotting was used to detect the effect of total flavonoids from Ampelopsis grossedentata on protein expressions of IRS-1/PI3K p85/AKT/GLUT4 signaling transduction.Results:When the total flavonoids from Ampelopsis grossedentata was 50μg/mL,the cell survival rate was the highest.According to the detection of C2C12 cell viability,the high,medium,and low concentrations of total flavonoids from Ampelopsis grossedentata used in subsequent cell experiments were 80μg/mL,50μg/mL,and 30μg/mL,respectively.Compared with the high glucose model group,the glucose consumption of total flavonoids from Ampelopsis grossedentata in the high,medium and low dose groups increased significantly(P<0.01).The high and medium dose groups of total flavonoids from Ampelopsis grossedentata significantly increased the protein expression of IRS-1/PI3K p85/AKT/GLUT4 signaling pathway(P<0.01).Conclusion:Total flavonoids of Ampelopsi

关 键 词：藤茶总黄酮 C2C12细胞 骨骼肌 胰岛素抵抗 IRS-1/PI3K p85/AKT/GLUT4信号通路 

分 类 号：R587.1[医药卫生—内分泌]