运脾活络汤通过调控 Zucker 糖尿病肥胖大鼠肝脏 miR-29a-3p 改善胰岛素抵抗
Yunpi Heluo decoction attenuates insulin resistance by regulating liver miR-29a-3p in Zucker diabetic fatty rats.
机构信息
Zhejiang Chinese Medical University School of Pharmacy, Hangzhou, 310053, China.
Endocrinology Department of Ningbo Municipal TCM Hospital, Ningbo, 315010, China.
出版信息
J Ethnopharmacol. 2019 Oct 28;243:111966. doi: 10.1016/j.jep.2019.111966. Epub 2019 May 23.
BACKGROUND AND OBJECTIVE
Yunpiheluo (YPHL) decoction is a Chinese herbal formula with unique advantages for the treatment of type 2 diabetes mellitus (T2DM). The aim of the present study was to investigate changes in miRNA expression and downstream gene expression in Zucker diabetic fatty (ZDF) rats treated with YPHL to determine whether YPHL could be used as an adjuvant treatment of T2DM.
METHODS
Serum and liver total cholesterol (TC) and triglycerides (TG) levels, insulin resistance index (IR) and differentially expressed miRNAs were detected in a T2DM ZDF rat model. miRNA target prediction was based on bioinformatic algorithms and dual luciferase reporter assay. Protein expression of genes in the insulin receptor signaling pathway was detected by Western blot. The IR cell model was established and the effects of lyophilized YPHL powder on the protein expressions were observed by transfecting specific miRNA mimics and inhibitors.
RESULTS
The miR-29a-3p expression level was significantly increased in the liver of ZDF rats. Insulin receptor substrate 1 (IRS1) was the target gene of miR-29a-3p. IRS1 mRNA and protein expressions of IRS1, IRS1 (phospho S307), protein kinase B (Akt), Akt (phosphor ser473) and pyruvate dehydrogenase lipoamide kinase isozyme 1 (PDK1) were decreased significantly. miR-29a-3p over-expression decrease IRS1 and the others protein expressions in the HepG2 IR cell model while anti-miR-29a-3p showed the opposite result. The miR-29a-3p level was decreased, and the expressions of IRS1 mRNA and the above proteins were all increased after YPHL treatment.
CONCLUSION
miR-29a-3p played a functional role in insulin receptor signaling in the liver of ZDF rats. YPHL decoction attenuated IR in T2DM probably by down-regulating or maintaining the miR-29a-3p level, increasing the expression of IRS1 mRNA and its phosphorylated proteins, and regulating the expression of insulin receptor signaling-related proteins. YPHL may prove to be an alternative treatment for T2DM.
背景与目的
运脾活络方（YPHL）是一种具有治疗 2 型糖尿病（T2DM）独特优势的中药复方。本研究旨在观察 YPHL 对 ZDF 糖尿病大鼠模型 miRNA 表达及下游基因表达的变化，探讨 YPHL 是否可作为 T2DM 的辅助治疗方法。
方法
通过检测 T2DM ZDF 大鼠模型血清和肝脏总胆固醇（TC）、三酰甘油（TG）水平、胰岛素抵抗指数（IR）以及差异表达 miRNA，分析 YPHL 对 ZDF 糖尿病大鼠的作用。利用生物信息学算法和双荧光素酶报告基因实验预测 miRNA 靶基因。采用 Western blot 检测胰岛素受体信号通路中基因的蛋白表达。建立胰岛素抵抗细胞模型，转染特异性 miRNA 模拟物和抑制剂，观察冻干 YPHL 粉末对细胞蛋白表达的影响。
结果
ZDF 大鼠肝脏中 miR-29a-3p 表达水平显著升高，胰岛素受体底物 1（IRS1）是 miR-29a-3p 的靶基因。IRS1mRNA 及其蛋白表达、IRS1（磷酸化 S307）、蛋白激酶 B（Akt）、Akt（磷酸化 ser473）和丙酮酸脱氢酶激酶同工酶 1（PDK1）均显著降低。在 HepG2 胰岛素抵抗细胞模型中，过表达 miR-29a-3p 可降低 IRS1 及其它蛋白的表达，而抑制 miR-29a-3p 则显示出相反的结果。YPHL 治疗后 miR-29a-3p 水平降低，IRS1mRNA 及其上述蛋白表达均增加。
结论
miR-29a-3p 在 ZDF 大鼠肝脏胰岛素受体信号通路中发挥功能作用。YPHL 可能通过下调或维持 miR-29a-3p 水平、增加 IRS1mRNA 及其磷酸化蛋白的表达以及调节胰岛素受体信号相关蛋白的表达，减轻 T2DM 大鼠的胰岛素抵抗。YPHL 可能成为 T2DM 的一种替代治疗方法。
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