Protective effects of rutin on liver injury in type 2 diabetic db/db mice
Protective effects of rutin on liver injury in type 2 diabetic db/db mice
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DOI：
10.1016/j.biopha.2018.08.046
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发表时间：
2018-11-01
影响因子：
7.5
通讯作者：
Gou, Xiangbo
中科院分区：
文献类型：
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作者：
Liang, Weishi;Zhang, Dandan;Gou, Xiangbo
本研究旨在评价芦丁对2型糖尿病小鼠肝脏的保护作用并探讨其相关机制。选取db/db小鼠作为2型糖尿病（T2DM）模型，其随机血糖（RBG）和口服葡萄糖负荷2小时后的血糖水平超过16.7 mmol/L。给予120 mg/kg或60 mg/kg芦丁后，检测不同组体内外RBG、口服糖耐量、血清丙氨酸氨基转移酶（ALT）和天冬氨酸氨基转移酶（AST）以及晚期糖基化终末产物（AGEs）。光镜和电镜观察肝脏病理变化。 Western blotting检测胰岛素受体底物2（IRS-2）蛋白表达量、p85上磷脂酰肌醇3激酶（PI3K）、Ser473上Akt、Ser9上糖原合酶激酶3β（GSK-3β）磷酸化情况，实时定量PCR检测IRS-2 mRNA表达量。此外，动态观察芦丁对非酶蛋白糖基化体系中AGEs生成的影响，采用Cell Counting Kit-8（CCK-8）法检测芦丁对HepG2肝细胞增殖活性的影响。结果显示，模型组小鼠口服葡萄糖耐量试验（OGTT）的RBG和葡萄糖水平显着高于正常组，经芦丁治疗后明显降低。芦丁可降低db/db小鼠血清中的ALT、AST活性和AGEs水平，增强db/db小鼠肝组织中IRS-2、P-PI3K(p85)、P-Akt(Ser473)、P-GSK-3β(Ser9)蛋白和IRS-2 mRNA的表达。此外，芦丁能显着缓解db/db小鼠肝脏结构紊乱，减少肝细胞变性、坏死和胶原纤维形成。体外实验结果还表明，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。总的来说，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产生来介导的。
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 3 beta (GSK-3 beta) 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-3 beta (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-3 beta signal pathway, promoting hepatocyte proliferation, reducing blood glucose level and generation of AGEs.