Effect of lncRNA-1700020I14Rikon the fibrosis in mouse mesangial cells in high glucose concentration / 基础医学与临床

ABSTRACT

Objective To construct the lncRNA-1700020I14Rik plasmid 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 1700020I14Rik in 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 analyze the expression 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.Conclusions The plasmidpcDNA3.1(+)-1700020I14Rik is able to effectively express the lncRNA-1700020I14Rik.Over-expression of 1700020I14Rik may protect mesangial cells from fibrosis conduced in high glucose medium.