SiRNA-HDAC5对非肥胖型糖尿病鼠组蛋白修饰异常的干预及其治疗作用  

Intervention and therapeutic effect of siRNA-HDAC5 on abnormal histone modification in non-obese diabetic mice

作　　者：欧阳琳[1] 王燕飞[2] 刘玲娇[2] 彭佑铭[3] 侯粲[1] 

机构地区：[1]中南大学湘雅二医院重症医学科,长沙410011 [2]中南大学湘雅二医院内分泌科糖尿病免疫学教育部重点实验室国家代谢性疾病临床医学研究中心,长沙410011 [3]中南大学湘雅二医院肾脏病研究所,长沙410011

出　　处：《中南大学学报（医学版）》2015年第5期464-470,共7页Journal of Central South University ：Medical Science

基　　金：国家自然科学基金青年基金(81200580);教育部博士点基金新教师类(20120162120090);湖南省自然科学基金(14JJ3042);中央高校青年教师助推专项资助(2012QNZT157);湖南省发改委基金(湘发改高技[2013]1199)~~

摘　　要：目的:通过小干扰RNA(small interfering RNA,si RNA)干预非肥胖型糖尿病(non-obese diabetic,NOD)鼠脾CD4+T细胞组蛋白去乙酰化酶5(histone deactylase 5,HDAC5)的表达,检测免疫、炎症及肾功能相关指标,评估si RNAHDAC5对NOD鼠的治疗作用,探讨HDAC5与糖尿病肾病之间的关系。方法:随机将NOD鼠分为3组,在12周龄时分别于尾静脉注射si RNA-HDAC5,si RNA-Control或生理盐水,于18,24和30周龄取样本检测,以血糖仪检测各组NOD鼠血糖,ELISA检测尿白蛋白排泄率及血清细胞因子IL-1,IL-6,IL-18和TNF-α水平。实时定量PCR检测各组NOD鼠CD4+T细胞CD11a,CCR5以及CX3CR1转录水平,Western印迹检测NOD鼠脾脏CD4+T细胞HDAC5蛋白水平。结果:与未干预对照组相比,HDAC5干预组小鼠血糖和尿白蛋白排泄率均显著降低,血清IL-1,IL-6,IL-18及TNF-α细胞因子水平下降,脾CD4+T细胞CD11a,CCR5以及CX3CR1明显降低。结论:阻断NOD鼠的HDAC5表达能减缓糖尿病鼠肾损伤的发生和发展。Objective: To evaluate therapeutic effect of si RNA-HDAC5 on non-obese diabetic(NOD) mice by using small interference RNA(si RNA) technique to knock down the expression of HDAC5 in spleen CD4+ T cells. Methods: NOD mice, 12-weeks old, were randomly divided into 3 groups and were given normal saline, si RNA-Control or si RNA-HDAC5 through caudal vein injection. The spleens and other samples were collected at the 18 th, 24 th or 30 th week. The blood glucose was tested by blood glucose meter. The urinary albumin and serum levels of IL-1, IL-6, IL-18, and TNF-α were detected by ELISA. The m RNA levels of CD11 a, CCR5, and CX3CR1 in spleen CD4+ T cells were measured by quantitative Real-time PCR. The HDAC5 protein level in spleen CD4+ T cell was detected by Western blot. Results: Compared with the control group, the si RNA-HDAC5 group showed a significant decrease in blood glucose, urine albumin excretion rate, serum cytokine and the m RNA levels of CD11 a, CCR5, and CX3CR1, consist with the decrease in protein level of HDAC5. Conclusion: Inhibition of HDAC5 expression in NOD mice could effectively alleviate the onset and development of kidney damage caused by diabetes.

关 键 词：组蛋白去乙酰化 组蛋白去乙酰化酶5 细胞因子 尿白蛋白/尿肌酐比率 

分 类 号：R587.1[医药卫生—内分泌] R589.2[医药卫生—内科学]