反义线粒体非编码RNA-2在高糖环境下的肾小球系膜细胞中的表达及作用机制

Expression and mechanism of Long non-coding RNA ASncmtRNA-2 in high glucose treated human renal mesangial cells

目的:观察长链非编码RNA反义线粒体非编码RNA-2(ASncmtRNA-2)在高糖环境下肾小球系膜细胞中的表达,探讨ASncmtRNA-2在氧化应激介导的糖尿病肾病纤维化中的作用。方法:采用实时荧光定量聚合酶链反应(qRT-PCR)法检测各组细胞ASncmtRNA-2、TGF-β1和FN mRNA表达。DCFH-DA法测定细胞内活性氧(ROS)相对水平。结果:与0h组比较,8h组ASncmtRNA-2mRNA的相对表达量无明显变化,但是从16h起,高糖组与高糖+L-NAME组ASncmtRNA-2 mRNA相对表达量显著高于低糖组(P均<0.05),高糖+L-NAME组ASncmtRNA-2mRNA相对表达量显著低于高糖组(P均<0.05)。高糖组细胞内ROS相对水平显著高于低糖组(P<0.05),高糖+LNAME组细胞内ROS相对水平显著低于高糖组(P<0.05)。高糖组细胞ASncmtRNA-2、TGF-β1和FN mRNA相对表达量显著高于低糖组(P均<0.05),高糖+L-NAME组ASncmtRNA-2、TGF-β1和FN mRNA相对表达量显著低于高糖组(P均<0.05)。高糖+ASncmtRNA-2siRNA组细胞ASncmtRNA-2、TGF-β1和FN mRNA相对表达量显著低于高糖组(P<0.05)。结论:ASncmtRNA-2在高糖环境下的肾小球系膜细胞中高表达,并且可能通过正调节促纤维化因子的表达来促进糖尿病肾病中的肾小球纤维化,这就为糖尿病肾病治疗提供新的靶点。

Objective：To observe the expression of Long non-coding RNA antisense mitochondrial non-coding RNA-2（ASncmtRNA-2）in high glucose（HG）treated human renal mesangial cells（HRMCs）and explore the role of ASncmtRNA-2 in oxidative stress mediated diabetic nephropathy（DN）fibrosis.Methods：The expression levels of ASncmtRNA-2,transforming growth factor β1（TGF-β1）and fibronectin（FN）mRNA in cultured HRMCs were measured by qRT-PCR.In addition,relative reactive oxygen species（ROS）levels in HRMCs were detected with the non-fluorescent probe DCFH-DA assays.Results：Compared with 0 h,the expression of ASncmtRNA-2 mRNA remained unchanged in all groups at 8 hpost treatment.However,the level of ASncmtRNA-2 mRNA was increased significantly in HG and HG＋NG-nitro-L-Arginine methylester（L-NAME）treated cells compared with low glucose（LG）treated cells from 16 honwards（P〈0.05）,while the level of ASncmtRNA-2 mRNA in the HG＋L-NAME group was decreased compared with the HG group（P〈0.05）.Moreover,ROS fluorescence was significantly up-regulated in HG-treated cells compared with LG-treated cells（P〈0.05）,while the ROS fluorescence in HG＋L-NAME group was suppressed compared with HG-treated cells（P〈0.05）.In addition,levels of ASncmtRNA-2,TGF-β1 and FN mRNA were significantly up-regulated in HG treated cells compared with LG treated cells（P〈0.05）while levels of ASncmtRNA-2,TGF-β1 and FN mRNA in HG＋L-NAME group were down-regulated compared with HG group（P〈0.05）.Finally,the expression of ASncmtRNA-2,TGF-β1 and FN mRNA were significantly decreased in HG＋ASncmtRNA-2 siRNA group compared with HG group（P〈0.05）.Conclusion：ASncmtRNA-2 is up-regulated in HG treated cells and may promote glomerular fibrosis in DN via positively regulating the expression of pro-fibrotic factors.These findings may provide novel potential treatments for DN.