Journal of Shandong University (Health Sciences) ›› 2020, Vol. 1 ›› Issue (7): 7-14.doi: 10.6040/j.issn.1671-7554.0.2019.1007

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miR-21-5p targeted TIMP3 to inhibit proliferation and extracellular matrix accumulation of mesangial cells in Type II diabetic nephropathy mice

ZHANG Baowen1, LEI Xiangli1, LI Jinna2, LUO Xiangjun1, ZOU Rong1   

  1. 1. Department of Nephrology, Affiliated Nanhua Hospital, University of South China, Hengyang 421002, Hunan, China;
    2. Department of Nephrology, Hengyang First Peoples Hospital, Hengyang 421002, Hunan, China
  • Online:2020-07-20 Published:2020-07-10

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Abstract: Objective To investigate the effects of miR-21-5p on the expression of tissue inhibitor of metalloproteinases-3(TIMP3)in kidney tissue of type 2 diabetic nephropathy(T2DN)mice, and the proliferation and extracellular matrix accumulation of mesangial cells. Methods Ten db/m mice were selected as control group, and other 30 SPF male db/db T2DN mice were randomly divided into model(T2DN)group, miR-21-5p agomir group and miR-21-5p antagomir group with 10 mice in each group. Normal saline, miR-21-5p agomir and miR-21-5p antagomir were injected respectively into tail vein once every three days, 7 times in total. The expression of miR-21-5p was detected by qRT-PCR in kidney tissues of mice. The expression levels of Upro/24 h, Scr and BUN in urine of mice were detected by ELISA assay. The kidney pathological changes were observed by HE staining. The extracellular matrix accumulation was observed by PAS staining. The expression levels of TIMP3, Col IV and FN protein in renal tissue were detected by Western blotting method. Dual luciferase reporter system was used to detect the targeting relationship between miR-21-5p and TIMP3. Results In the T2DN group and miR-21-5p agomir group, the levels of Upro/24 h(t=84.67, P<0.001; t=100.44, P<0.001), Scr(t=16.81, P<0.001; t=36.76, P<0.001), and BUN(t=19.26, P<0.001; t=52.42, P<0.001)increased, mesangial proliferated, basement membrane thickened, relative area of mesangial matrix increased(t=9.10,P<0.001;t=14.16,P<0.001), TIMP3 expression was abnormally lower(t=8.51,P=0.001;t=12.66,P<0.001), and fibrosis protein Col IV expression(t=10.04,P<0.001;t=23.54,P<0.001)and FN expression(t=11.49,P<0.001;t=22.34,P<0.001)was abnormally higher than those in the control group. Compared with T2DN group, the levels of Upro/24 h, Scr and BUN decreased(tUpro/24 h=20.31,P<0.001;tScr=7.902,P<0.001;tBUN=8.913,P<0.001), mesangial hyperplasia and basement membrane thickening alleviated, the relative area of mesangial matrix decreased(t=7.96,P=0.001), TIMP3 expression increased(t=11.71,P<0.001), fibrosis proteins Col IV expression(tCol IV=6.58,P=0.003)and FN expression(tFN=6.27,P=0.003)decreased in the miR-21-5p antagomir group. The biochemical indexes and symptom of glomerular diseases in miR-21-5p antagomir group and miR-21-5p agomir group were opposite. Dual luciferase assay showed that miR-21-5p could target the expression of TIMP3 gene. Conclusion miR-21-5p targeting TIMP3 inhibits the proliferation of mesangial cells and the extracellular matrix accumulation in T2DN mice.

Key words: miR-21-5p, Tissue inhibitor of metalloproteinases-3, Type II diabetic nephropathy, Cell proliferation, Extracellular matrix

CLC Number: 

  • R574
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