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目的:研究肝脏组蛋白H3表观修饰的变化在2型糖尿病小鼠发病过程中的作用,探讨其临床意义。方法:30只C57BL/6J小鼠分为正常组、高脂高糖组和糖尿病组,正常组和高脂高糖组小鼠分别给予正常饲料和高脂高糖饲料,糖尿病模型采用高脂高糖饲料联合注射链脲佐菌素(STZ)方法构建。采用HE染色、免疫印迹法、实时定量PCR和ChIP技术分别检测小鼠肝脏病理学变化、肝脏中总组蛋白H3的修饰状况及与糖代谢相关基因丙酮酸激酶(Pklr)、葡萄糖转运蛋白2(Glut2)、葡糖糖激酶(Gck)、过氧化物酶体增生物激活受体γ辅助活化因子1(Ppargc1a)和胰岛素受体(Insr)的表达水平和基因启动子区组蛋白的修饰变化。结果:糖尿病组小鼠肝脏病理学(HE染色)和组蛋白修饰模式均发生了明显变化。与正常组比较,高脂高糖组小鼠H3K23的乙酰化下降(P<0.01),H3K9Ac和H3K9Me2修饰水平上调(P<0.01),H3K4Me保持不变(P>0.05),糖代谢相关基因Pklr、Glut2和Gck的表达水平升高(P<0.01);糖尿病组小鼠H3K23Ac和H3K9Ac修饰水平下降(P<0.05或P<0.01),H3K9Me2和H3K4Me的修饰水平升高(P<0.01),糖代谢相关基因Pklr、Glut2、Gck、Ppargc1a和Insr表达水平明显下调(P<0.05或P<0.01)。Pklr、Glut2启动子区H3K23Ac、H3K9Ac、H3K9Me2和H3K4Me修饰水平变化与基因的表达水平变化相吻合。结论:肝脏组蛋白表观修饰变化可能参与了2型糖尿病的发生发展。
OBJECTIVE: To study the role of changes in histone H3 histone modification in the pathogenesis of type 2 diabetic mice and its clinical significance. Methods: Thirty C57BL / 6J mice were divided into normal group, high-fat and high-sucrose group and diabetic group. The normal group and high-fat and high-sucrose group were given normal diet and high-fat and high-sucrose diet respectively. Construction of sugar feedstains in combination with streptozotocin (STZ) injection. HE staining, Western blot, real-time quantitative PCR and ChIP were used to detect the changes of liver pathology, the modification of total histone H3 in liver and the relationship between Pklr and glucose transporter 2 Glut2, Gck, Ppargc1a and Insr and histone modifications of the promoter region of the gene. Results: Hepatic pathology (HE staining) and histone modifications in diabetic mice all changed significantly. Compared with the normal group, the acetylation of H3K23 was decreased (P <0.01), the changes of H3K9Ac and H3K9Me2 were up-regulated (P <0.01), H3K4Me was unchanged (P> 0.05), Pklr , Glut2 and Gck (P <0.01). The levels of H3K23Ac and H3K9Ac in diabetic mice decreased (P <0.05 or P <0.01), the levels of H3K9Me2 and H3K4Me increased (P <0.01) Metabolic related genes Pklr, Glut2, Gck, Ppargc1a and Insr expression levels were significantly down-regulated (P <0.05 or P <0.01). Pklr, Glut2 promoter H3K23Ac, H3K9Ac, H3K9Me2 and H3K4Me modification level changes consistent with changes in gene expression levels. Conclusion: The changes of histone modifications in liver may be involved in the development of type 2 diabetes mellitus.