Role Of Long Non-coding RNA Uc.412 In Renal Fibrosis

Objective Chronic kidney disease threatens human health for a long time.However,the mechanism of renal fibrosis,the key pathological process of chronic kidney disease,has not been clear,and there is no effective target to inhibit fibrosis.In recent years,long non-codingRNA(LncRNA)has been a research hotspot as a key molecule that can regulate transcription,translation and other key functions.Previous studies of our group found that LncRNA uc.412 may be related to renal fibrosis.This experiment was to explore the expression and possible mechanism of LncRNA uc.412 in renal fibrosis.Methods 12 male C57 BL / 6 mice were randomly divided into two groups:control group(n=6)and unilateral ureteral obstruction(UUO)group(n=6).In UUO group,we separated and ligated the ureter in mice to make it obstructed and induce them to be a renal fibrosis model,while control group were treated with blunt ureteral separation but without ligation.The mice were sacrificed at 14 days after operation.We collected blood and renal tissue samples,and then measured serum creatinine and urea nitrogen.Besides,Masson staining was used to test the degree of renal fibrosis.The expression of TGF-β1,LncRNA uc.412,COL1 and α-SMA in renal tissues were detected by q PCR.The expression of COL1 was detected by Western Blot.Rat mesangial cells were cultured in vitro and treated with TGF-β 1 and siRNA.The expression level of LncRNA uc.412 was detected by q PCR.The expression level of α – SMA was detected by Western Blot.Rat mesangial cells were cultured in vitro and treated with TGF-β 1 and Smad3 inhibitor SIS3.We detected the level of LncRNA uc.412 by q PCR.Results In vivo study,compared with control group,the level of biochemical indexes(serum creatinine and urea nitrogen)of UUO group was significantly increased(P<0.05).Masson staining showed the degree of fibrosis in UUO group was significantly aggravated than control group.Compared with control group,the expression levels of LncRNA uc.412,COL1,α-SMA,and TGF-β1 in UUO group were significantly increased(P<0.05).In vitro study,compared with control group,the expression of COL1 protein in UUO group was significantly increased(P<0.05).The expression of LncRNA uc.412 in TGF-β 1 group was higher than that in control group,while the expression of LncRNA uc.412 in TGF-β 1 + 25 nmol / L siRNA group,TGF-β 1 + 50 nmol / L siRNA group and TGF-β 1 + 75 nmol / L siRNA group was lower than that in TGF-β 1 group(P<0.05).Compared with TGF-β 1 group,TGF-β 1 + siRNA group can significantly reduce the expression level of LncRNA uc.412(P < 0.05).Compared with TGF-β 1 group,TGF-β 1 + siRNA group could significantly reduce the expression level of α-SMA.Compared with control group,the level of the expression of LncRNA uc.412 in the TGF-β1 group was significantly increased(P<0.05),while it was significantly decreased in the SIS3 group compared with the TGF-β1 group(P<0.05).Conclusions The increased expression of LncRNA uc.412 may be involved in the development of renal fibrosis in UUO model.TGF-β1 may induce the increased expression of LncRNA uc.412 in mesangial cells through TGF-β1/Smad3 signaling pathway,and then induce fibrosis.