Clinical Medical College, North China University of Science and Technology, Tangshan 063210, PR China; Basic Medical College, North China University of Science and Technology, Tangshan 063210, PR China.
Basic Medical College, North China University of Science and Technology, Tangshan 063210, PR China; Department of Pharmacology, North China University of Science and Technology, Tangshan 063210, PR China.
Biomed Pharmacother. 2018 Nov;107:721-728. doi: 10.1016/j.biopha.2018.08.046. Epub 2018 Aug 20.
The aim of this study was to evaluate the protective effect of rutin on the liver of type 2 diabetic mice and explore the correlation mechanism. The db/db mice, selected as the type 2 diabetes mellitus (T2DM) models, have random blood glucose (RBG) and glucose level after 2 h of oral glucose loading of more than 16.7 mmol/L. After administration of 120 mg/kg or 60 mg/kg rutin, to T2DM mice, RBG, oral glucose tolerance, alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in serum, and advanced glycation end products (AGEs) in vivo and vitro of different groups were detected. The liver pathological changes were observed under light and electron microscopy. Western blotting was used to detect the protein expression of insulin receptor substrate 2 (IRS-2) and phosphorylation of phosphatidylinositol 3 kinase (PI3K) on p85, Akt on Ser473, glycogen synthase kinase 3β (GSK-3β) on Ser9, real-time quantitative PCR was used to detect IRS-2 mRNA expression. Moreover, dynamically observing the effect of rutin on the generation of AGEs in non-enzymatic protein glycosylated system, Cell Counting Kit-8 (CCK-8) method was used to detect the effect of rutin on proliferation activity of HepG2 liver cells. The results showed that RBG and glucose levels of oral glucose tolerance test (OGTT) of mice in model group were significantly higher than that of normal group, which were significantly reduced after the rutin treatment. Rutin could reduce the ALT, AST activities and AGEs level in serum and potentiate the expression of IRS-2, P-PI3K (p85), P-Akt (Ser473), P-GSK-3β (Ser9) protein and IRS-2 mRNA in the liver tissue of db/db mice. Moreover, rutin could significantly alleviate the structure disorder of liver, reduce the degeneration and necrosis of liver cells and formation of collagen fibers of db/db mice. The results in vitro also showed that rutin could obviously inhibit the generation of AGEs, and promoted the proliferation activity of high glucose-stimulating HepG2 cells. In general, the protective effect of rutin on the liver of T2DM may be mediated by facilitating signal transduction and activated state of insulin IRS-2/PI3K/Akt/GSK-3β signal pathway, promoting hepatocyte proliferation, reducing blood glucose level and generation of AGEs.
本研究旨在评估芦丁对 2 型糖尿病小鼠肝脏的保护作用,并探讨其相关机制。选择 db/db 小鼠作为 2 型糖尿病(T2DM)模型,其随机血糖(RBG)和口服葡萄糖耐量 2 小时后血糖水平均高于 16.7mmol/L。给予 120mg/kg 或 60mg/kg 芦丁后,检测 T2DM 小鼠体内外 RBG、口服葡萄糖耐量、血清丙氨酸氨基转移酶(ALT)和天冬氨酸氨基转移酶(AST)、晚期糖基化终产物(AGEs)的变化。在光镜和电镜下观察肝组织的病理变化。采用 Western blot 检测胰岛素受体底物 2(IRS-2)和磷酸化磷脂酰肌醇 3 激酶(PI3K)p85、Akt Ser473、糖原合成酶激酶 3β(GSK-3β)Ser9 蛋白的表达,实时定量 PCR 检测 IRS-2mRNA 的表达。此外,采用细胞计数试剂盒-8(CCK-8)法检测芦丁对 HepG2 肝细胞增殖活性的影响,动态观察芦丁对非酶蛋白糖基化体系中 AGEs 生成的影响。结果表明,模型组小鼠 RBG 和口服葡萄糖耐量试验(OGTT)血糖水平明显高于正常组,芦丁治疗后明显降低。芦丁能降低血清 ALT、AST 活性和 AGEs 水平,增强 db/db 小鼠肝组织 IRS-2、P-PI3K(p85)、P-Akt(Ser473)、P-GSK-3β(Ser9)蛋白和 IRS-2mRNA 的表达。此外,芦丁能明显改善肝组织结构紊乱,减轻 db/db 小鼠肝细胞变性坏死和胶原纤维形成。体外实验结果也表明,芦丁能明显抑制 AGEs 的生成,促进高糖刺激的 HepG2 细胞的增殖活性。综上所述,芦丁对 T2DM 肝脏的保护作用可能是通过促进胰岛素 IRS-2/PI3K/Akt/GSK-3β 信号通路的信号转导和激活状态,促进肝细胞增殖,降低血糖水平和 AGEs 生成来实现的。