关键词: 长链非编码RNA, 表达质粒, 糖尿病, 糖尿病肾病

Abstract: Objective To construct the lncRNA-1700020I14Rikplasmid and detect its effect on the fibrosis of mice mesangial cell (MMC) cultured with high glucose medium. Methods RT-qPCR was used to measure the expression of 1700020I14Rikin MMC cultured with low glucose medium or high glucose. Total RNA was extracted from MMC and cDNA was got by RT-PCR. The whole fragment of lncRNA-1700020I14Rik amplified by PCR was constructed into plasmid pcDNA3.1(+) through PCR. Lipidosome 3000 was used to transfect the plasmid into the MMC cultured with high glucose medium and RT-qPCR was used to measure the expression level of 1700020I14Rik. Western blot was used to analysis the expressions of fibronectin, collagen Ⅳ and TGF-β1. Results 1700020I14Rik was significantly down-regulated in MMC cultured with high glucose and it was significantly up-expressed in the MMC after transfecting with pcDNA3.1(+)-1700020I14Rik. The expressions of fibronectin, collagen Ⅳ and TGF-β1 were down-regulated by 1700020I14Rik. Conclusion The plasmid pcDNA3.1(+)-1700020I14Rikis able to effectively express the lncRNA-1700020I14Rik. Over-expression of 1700020I14Rik may protect mesangial cells from fibrosis conduced under high glucose condition.

Key words: Long non-coding RNA, Expression plasmid, Diabetes mellitus, Diabetic nephropathy