MiroRNA-21 mediates myocardial fibroblasts paracrine stimulated by indoxyl sulfate in vitro
10.3760/cma.j.issn.1001-7097.2017.12.009
- VernacularTitle:硫酸吲哚酚通过上调微小RNA-21表达参与心肌成纤维细胞的旁分泌
- Author:
Jianguang GONG
1
;
Yiwen LI
;
Juan JIN
;
Li ZHAO
;
Yiqiao LI
;
Qiang HE
Author Information
1. 浙江省人民医院杭州医学院附属人民医院肾脏病科
- Keywords:
Uremia;
MicroRNA;
Paracrine communication;
Indican
- From:
Chinese Journal of Nephrology
2017;33(12):935-939
- CountryChina
- Language:Chinese
-
Abstract:
Objective To explore the expression of miroRNA-21 (miRNA-21) in myocardial fibroblasts stimulated by indoxyl sulfate (IS) and its role on paracrine factors of myocardial fibroblasts.Methods Myocardial fibroblasts which derived from C57BL/6J mice were divided into control group and IS group,and their expressions of miRNA-21 were detected by real time PCR after 48 h.MiRNA-21 inhibitor transfection was applied to silence miRNA-21 expression.Myocardial fibroblasts were divided into creatinine (Scr) group (Scr treated for 48 h),Scr+IS group (Scr and 50 μmol/L IS treated for 48 h),Scr+miRNA-21 inhibitor group (miRNA-21 inhibitor treated for 24 h and then Scr treated for 48 h) and Scr+miRNA-21 inhibitor+ IS group (miRNA-21 inhibitor pretreated for 24 h and then Scr and IS treated for 48 h).Enzyme-linked immunosorbent assay was performed to evaluate the expressions of interleukin-1 (IL-1),interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α).Western blotting was applied to detect the expressions of transforming growth factor-β (TGF-β),matrix metalloproteinases 2 (MMP2),matrix metalloproteinases 9 (MMP9) and tissue inhibitor of metalloproteinases 1 (TIMP1).Results The expression of miRNA-21 was obviously increased in IS group than that in control group (P ＜ 0.01).Compared with those in Scr group,the expressions of IL-1,IL-6,TNF-α,TGF-β,MMP2 and MMP9 significantly increased (all P ＜ 0.05),while the expression of TIMP1 decreased (P ＜ 0.05).When the expression of miRNA-21 was inhibited,the expressions of IL-1,IL-6,TNF-α,TGF-β and MMP2 in Scr+miRNA-21 inhibitor+IS group significantly decreased than those in Scr+IS group (all P ＜0.05),and the expression of TIMP1 decreased further (P ＜ 0.05).Conclusions IS can promote the expression of miRNA-21 in myocardial fibroblasts.MiRNA-21 plays an important role in regulating inflammatory factors and pro-fibrogenic cytokines in myocardial fibroblasts.
