黄酮衍生物C45对高脂诱导肥胖小鼠的糖脂代谢改善作用及机制探讨

Effect and mechanism of flavonoid derivative C45 on the metabolism of glucose and lipid in high fat diet-induced obese mice

目的探讨黄酮衍生物C45对高脂诱导肥胖小鼠的糖脂代谢改善作用及机制。方法高脂饮食喂养建立胰岛素抵抗的肥胖小鼠模型,缺氧条件下共培养的脂肪细胞和巨噬细胞的条件培养基(HF/M)诱导AML12肝细胞胰岛素抵抗。给予肥胖小鼠和胰岛素抵抗的肝细胞黄酮衍生物C45处理,小鼠血清糖脂代谢和氧化应激指标分别用相应的试剂盒检测,流式细胞术检测小鼠外周血炎性细胞比例,天狼星红染色检测小鼠肝组织纤维化,免疫印迹法检测糖脂代谢和炎症相关分子的蛋白和磷酸化水平,荧光定量聚合酶链反应检测肝细胞脂代谢相关分子的mRNA水平。结果黄酮衍生物C45显著降低肥胖小鼠血清空腹血糖、空腹胰岛素、甘油三酯、总胆固醇、游离脂肪酸和丙二醛的水平,升高肥胖小鼠高密度脂蛋白-胆固醇/低密度脂蛋白-胆固醇比值和超氧化物歧化酶水平,降低外周血炎性中性粒细胞比例、肝组织胶原纤维数量和体积。黄酮衍生物C45逆转HF/M降低AML12细胞蛋白激酶B胰岛素敏感性的作用、降低过氧化物酶体增殖物激活受体γ和固醇调节元件结合蛋白1c的mRNA水平以及核因子-κB的磷酸化水平。结论黄酮衍生物C45调节糖脂代谢、氧化应激和炎症,改善胰岛素抵抗,具有抗糖尿病作用。

Objective To explore the effect and mechanism of flavonoid derivative C45(C45)on the metabolism of glucose and lipid in high fat diet-induced obese mice.Methods Insulin resistant obese mice model was induced by high fat diet(HFD)feeding.The condition medium from co-cultured adipocytes and macrophages under hypoxic condition was used to induce AML12 hepatocytes insulin resistance.Obese mice and insulin resistant hepatocytes were treated with C45.Glucose and lipid metabolism and oxidative stress related markers in mice serum were detected by commercial kits,respectively.The proportion of mice peripheral inflammatory cells was detected by flow cytometry.Mice liver fibrosis was detected by sirius red stain.The protein and phosphorylation levels of hepatic glucose metabolism-and inflammation-related molecules were detected by Western blot.The mRNA expressions related to lipid metabolism were measured by fluorescence quantitative PCR.Results C45 significantly decreased the serum levels of fasting glucose,insulin,TC,TG,NEFA and MDA,whereas increased levels of HDL/LDL and SOD of obese mice.C45 decreased the proportion of peripheral inflammatory neutrophils,the number and volume of collagen fibers in liver.C45 reversed HF/M-reduced insulin phosphorylated Akt level and increased the mRNA expressions of PPARγand SREBP1c in AML12 cells,as well as NF-κB phosphorylation.Conclusion Tiliroside derivative C45 regulates glucose and lipid metabolism,oxidative stress,inflammation and insulin resistance,which may prevent diabetes.