微阵列法评价丹蛭降糖胶囊对糖尿病大鼠棕色脂肪miRNAs表达谱的影响

Effect of Danzhi Jiangtang Capsule on Expression of miRNAs in Brown Adipose Tissue of Hyperglycemia Rats Based on Microarray Analysis

目的:基于微阵列分析法研究益气活血中药丹蛭降糖胶囊对糖尿病大鼠棕色脂肪mi RNA表达谱的影响。方法:将30只大鼠随机分为正常对照组、模型组、丹蛭降糖胶囊组,每组各10只。动物造模及给药结束后采集3组大鼠的棕色脂肪组织,并提取mi RNA,分别与mi RNA芯片进行杂交,进一步扫描荧光信号,对模型组、丹蛭胶囊组与正常对照组进行差异基因的筛选;并采用实时定量RT-PCR法验证部分差异表达的mi RNA。结果:此次实验丹蛭降糖胶囊组共筛选出17个差异基因;其中上调的有mi RNA-10b、mi RNA-26a、mi RNA-99a、mi RNA-196a、mi RNA-375、mi RNA-101b、mi RNA-151、mi RNA-152、mi RNA-98、mi RNA-328;下调的有mi RNA-103、mi RNA-27b、mi RNA-29a、mi RNA-133、mi RNA-155、mi RNA-143,荧光定量RT-PCR结果与mi RNAs芯片的相符合。Pathway分析显示,差异基因参与2条与血糖调节密切相关的通路。结论:具有益气活血功效的丹蛭降糖胶囊可通过影响棕色脂肪组织的mi RNA表达,减轻胰岛B细胞负担,改善胰岛素敏感性,从而调控血糖水平。

Objective：To evaluate the effect of Danzhi Jiangtang capsule on expression of miRNAs in brown adipose tissue（BAT） of hyperglycemia rats base on microarray analysis. Methods ： 30 healthy rats were randomly allocated into three groups as control group, model group, and Danzhi Jiangtang capsule group, with ten rats in each group. Brown adipose tissue were collected after model establish- ment and drug administration, extracted the miRNA, made hybridization with the miRNA chip respectively, scanned the fluorescent sig- nals and screened out differentially expressed genes among three groups, partial differential expression of miRNA were verified by RT- PCR. Results ： A total of 17 differentially expressed genes were screened, miRNA-10b, miRNA-26a, miRNA-99a, miRNA-196a, miRNA- 375, miRNA- 10 1b, miRNA- 151, miRNA- 152, miRNA-98, miRNA-328 were up-regulated; while miRNA- 103, miRNA-27b, miRNA-29a, miRNA-133,miRNA-155,miRNA-143 were down-regulated in the Danzhi Jiangtang capsule group, applied to the target gene analysis and RT-PCR verification, and found the similar result. The data analyzed by Pathway software indicated that the differentially expressed genes participated in two signal paths which closely related to blood glucose regulation. Conclusion：Danzhi Jiangtang capsule can regulate the expression of miRNAs in brown adipose tissue, reduce the burden of islet B cells, improve insulin sensitivity, consequently, adjust and control the blood glucose.