两色金鸡菊黄酮成分对高糖高脂诱导的细胞模型能量代谢紊乱的影响

目的观察两色金鸡菊黄酮成分对高糖高脂诱导大鼠肾小球系膜细胞HBZY-1能量代谢紊乱的影响,探讨其保护作用机制。方法采用高糖高脂诱导HBZY-1细胞建立体外糖尿病肾病代谢紊乱细胞模型,将HBZY-1细胞分为正常组、模型组和两色金鸡菊黄酮成分不同剂量组。采用MTS实验检测模型细胞增殖水平,倒置显微镜观察模型细胞形态变化,细胞划痕实验观察模型细胞细胞外基质分泌,Westernblot检测细胞能量代谢调控因子5’腺苷-磷酸活化蛋白激酶α(p-AMPKα)、乙酰辅酶a羧化酶1(ACC1)、甾醇调节元件结合蛋白1(SREBP1)及氧化应激因子一氧化氮合酶(e NOS)、尼克酰胺腺嘌呤二核苷酸磷酸氧化酶4(NOX4)的蛋白表达。结果两色金鸡菊黄酮成分能抑制高糖高脂诱导的HBZY-1细胞增殖,细胞间隙增大、密度显著降低;两色金鸡菊黄酮成分促进模型细胞p-AMPKα、p-ACC1、e NOS的蛋白表达,抑制模型细胞SREBP1、NOX4、ACC1的蛋白表达;划痕实验结果显示,两色金鸡菊黄酮成分对模型细胞细胞外基质分泌有显著抑制作用,同时α-平滑肌肌动蛋白的蛋白表达显著降低。结论两色金鸡菊黄酮成分可能通过调控AMPK/SREBP1/ACC1能量代谢通路,抑制模型细胞增殖、氧化应激,进而改善肾脏纤维化病变。

Effects of Flavonoids of Coreopsis tinctoria Nutt. on Energy Metabolism Disorder of High Glucose and High Fat-induced Cell Model

Objective To observe the effects of flavonoids of Coreopsis tinctoria Nutt. on energy metabolism disorder of high glucose and high fat-induced cell model;To explore its protective mechanism. Methods Rat diabetic nephropathy metabolic disorder cell model was established by high glucose and high fat-induced HBZY-1 cell. HBZY-1 cells were divided into normal group, model group and flavonoids of Coreopsis tinctoria Nutt. different dosage groups. MTS assay was used to detect the cell proliferation level of the model cells, and the morphology of the cells was observed under an inverted microscope. Cell scratch assay was used to observe the secretion of extracellular matrix in model cells. Western blot was used to detect the protein expressions of cellular energy metabolism regulator factor p-AMPKα, ACC1, SREBP1 and oxidative stress factor e NOS, and NOX4. Results Flavonoids of Coreopsis tinctoria Nutt. could inhibit cell proliferation of high glucose and high fat-induced HBZY-1 cells. Bigger cell space and lower cell density were detected after Flavonoids of Coreopsis Tinctoria Nutt treatment;FC could increase the expressions of p-AMPKα, p-ACC1, and eNOS and inhibit the expressions of SREBP1, NOX4, ACC1, and α-SMA;Cell scratch assay results showed that flavonoids of Coreopsis tinctoria Nutt. significantly inhibited ECM excretion, and α-SMA protein expression decreased. Conclusion Flavonoids of Coreopsis tinctoria Nutt. can inhibit cell proliferation and oxidative stress via regulating AMPK/SREBP1/ACC1 signaling pathway then improve the renal fibrosis.