滋肾丸对KKay小鼠肝脏胰岛素抵抗以及PI3K-Akt-FOXO1通路的影响  

Effects of Zishen pill on hepatic insulin resistance and PI3K-AKT-FOXO1 pathway in KKay mice

作　　者：陈源 吴悠 吴丽丽[2] 秦灵灵 刘铜华[2] CHEN Yuan;WU You;WU Lili;QIN Lingling;LIU Tonghua(Dongfang Hospital of Beijing University of Chinese Medicine,Beijing 100078,China)

机构地区：[1]北京中医药大学东方医院内分泌科,100078 [2]北京中医药大学教育部中医养生学重点实验室,100078 [3]北京中医药大学科技处,100078

出　　处：《环球中医药》2024年第3期377-383,共7页Global Traditional Chinese Medicine

基　　金：中医药防治糖尿病国际联合研究中心(2015B01022)。

摘　　要：目的 探究不同剂量滋肾丸水提物对于糖尿病小鼠降糖效果及该药对肝脏胰岛素抵抗的作用机制。方法 21只KKay小鼠随机分为模型组、滋肾丸高剂量组和滋肾丸低剂量组,每组7只;7只C57BL/6J小鼠作为正常组。连续灌胃给药6周,滋肾丸高剂量组予2.8 g/(kg·d),滋肾丸低剂量组予1.4 g/(kg·d),模型组、正常组予等体积蒸馏水。观察每周空腹血糖(fasting blood glucose, FBG),最后一周进行口服葡萄糖耐量试验(oral glucose tolerance test, OGTT),酶联免疫吸附法(enzyme-linked immunosorbent assay, ELISA)检测各组小鼠血清中的空腹胰岛素(fasting insulin, FINS)并计算胰岛素抵抗指数(homeostasis model assessment-insulin resistance, HOMA-IR),qPCR法检测胰岛素受体底物(insulin receptor substrate, IRS)、磷脂酰肌醇3-激酶(phosphoinositide 3-kinase, PI3K)、蛋白激酶B(protein kinase B,PKB/Akt)、叉头框蛋白1(forkhead box O1,FOXO1)、磷酸烯醇丙酮酸羧激酶(phosphoenolpyruvate carboxykinase, PEPCK)、葡萄糖-6-磷酸酶(glucose-6-phosphatase, G6pase)、葡萄糖激酶(glucokinase, GCK)基因表达水平;苏木素-伊红(hematoxylin-eosin staining, HE)染色和糖原过碘酸雪夫染色(periodic Acid-Schiff staining, PAS)观察肝脏病理情况和糖原分布。结果 与模型组相比,滋肾丸高剂量组和滋肾丸低剂量组小鼠FBG、HOMA-IR、OGTT曲线下面积明显下降,肝脏形态学改变部分减少,脂滴减少,糖原分布增加,FOXO1、PEPCK、G6pase基因表达均明显下降,滋肾丸高剂量组GCK基因表达水平显著上升,差异有统计学意义。结论 滋肾丸对于KKay小鼠降糖效果显著,能明显改善肝脏胰岛素抵抗,其机制可能是降低FOXO1、PEPCK、G6pase的转录水平,提高GCK的转录水平,从而抑制糖异生,促进糖原合成有关。Objective To evaluate the hypoglycemic effect of the traditional Chinese medicine decoction Zishen pill on diabetic mice and the mechanism of the drug on hepatic insulin resistance.Methods 21 KKay mice were randomly divided into Zishen pill high-dosage group and Zishen pill low-dosage group,a model group,and 7 C57 mice as the normal group.After continuous administration for 6 weeks,the fasting blood glucose was observed weekly,and the oral glucose tolerance test(OGTT)was performed in the last week of treatment.The enzyme-linked immunosorbent assay(ELISA)was used to detect the fasting insulin(FINS)in the serum of the mice in each group and homeostasis model assessment-insulin resistance(HOMA-IR)was calculated.RT-qPCR was applied to detect the mRNA expression of insulin receptor substrate(IRS),phosphoinositide 3-kinase(PI3K),protein kinase B(PKB/AKT),forkhead box O1(FOXO1),phosphoenolpyruvate carboxykinase(PEPCK),glucose-6-phosphatase(G6pase),glucokinase(GCK).Hematoxylin-eosin staining(HE)and periodic acid Schiff staining(PAS)were used to observe the liver morphology and glycogen distribution.Results Compared with the C group,the FBG,HOMA-IR,and area under curve(AUC)of OGTT in groups H and L decreased significantly,the changes in liver morphology of C group was partially diminished in Zishen pill treated groups,with fewer lipid droplets and more glycogen distribution.The gene expression of FOXO1,PEPCK,and G6pase decreased significantly,and the expression of GCK gene in group H increased significantly.Conclusion The Zishen pill has obvious hypoglycemic effect on KKay mice,and can significantly improve hepatic insulin resistance.The mechanism may be due to its effect of reducing the transcription of FOXO1,PEPCK,G6pase and increase the transcription of GCK,thereby inhibiting gluconeogenesis and promoting glycogen synthesis.

关 键 词：滋肾丸 小鼠 肝胰岛素抵抗 磷脂酰肌醇3-激酶通路 糖代谢 

分 类 号：R285.5[医药卫生—中药学]