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

目的：通过小干扰RNA(small interfering RNA，siRNA)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siRNAHDAC5 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siRNA-HDAC5，siRNA-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。

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

Objective: To evaluate therapeutic eff ect of siRNA-HDAC5 on non-obese diabetic (NOD) mice by using small interference RNA (siRNA) 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, siRNA-Control or siRNA-HDAC5 through caudal vein injection. The spleens and other samples were collected at the 18th, 24th or 30th 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 mRNA levels of CD11a, 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 siRNA-HDAC5 group showed a significant decrease in blood glucose, urine albumin excretion rate, serum cytokine and the mRNA levels of CD11a, CCR5, and CX3CR1, consist with the decrease in protein level of HDAC5. Conclusion: Inhibition ofHDAC5 expression in NOD mice could effectively alleviate the onset and development of kidney damage caused by diabetes.