circ_WBSCR17 attenuates high glucose-induced fibrosis and inflammation in human mesangial cells by regulating the miR-30a-5p /JAK1 axis / 安徽医科大学学报

ABSTRACT

Objective @# To investigate the influences of circ_WBSCR17 on high glucose-induced fibrosis and inflammation in human mesangial cells by regulating the miR-30a-5p /JAK1 axis．@*Methods @#Human mesangial cells HMCL were grouped into : NG group (5．5 mmol / L glucose-treated HMCL cells) ，HG group (30 mmol / L glucose- treated cells) ，si-NC group (30 mmol / L glucose + transfected with si-NC) ，si-circ_WBSCR17 group (30 mmol / L glucose + transfected with si-circ _ WBSCR17 ) ，si-circ _ WBSCR17 + inhibitor-NC group ( 30 mmol / L glucose + co-transfected with si-circ_WBSCR17 and inhibitor-NC) ，and si-circ_WBSCR17 + miR-30a-5p inhibitor group (30 mmol / L glucose + co-transfected with si-circ_WBSCR17 and miR-30a-5p inhibitor) ; RT-qPCR was performed to detect the expression of circ_WBSCR17 and miR-30a-5p in cells ; CCK-8 assay was performed to detect cell prolifer- ation ; flow cytometry was performed to detect apoptosis ; ELISA was performed to detect the expression levels of tumor necrosis factor-α (TNF-α) ，interleukin (IL) -6 and IL-8 ; Western blot was performed to detect the expression of JAK1，proliferating cell nuclear antigen ( PCNA) ，Bax，transforming growth factor-β1 ( TGF-β1 ) ，fibronectin (FN) ，collagen IV，and α-smooth muscle actin ( α-SMA) ; distribution of WBSCR17 was detected by fluorescence in situ hybridization (FISH) ; dual-luciferase reporter gene experiment was performed to verify the relationship between circ _ WBSCR17 and miR-30a-5p，miR-30a-5p and JAK1，respectively. @*Results @#Compared with the NG group，the HMCL cell proliferation ability of the HG group decreased，the levels of TNF-α , IL-6 and IL-8，the pro- tein expressions of p-JAK1 /JAK1，p-STAT1 / STAT1，p-STAT3 / STAT3，TGF-β1，FN，collagenIV and α-SMA，and the apoptosis ability increased (P＜0. 05) ; compared with HG group and si-NC group，the expression of miR-30a- 5p，OD450 value and PCNA expression in HMCL cells of si-circ_WBSCR17 group increased，the levels of TNF-α , IL- 6 and IL-8，the expressions of circ_WBSCR17，p-JAK1 /JAK1，p-STAT1 / STAT1，p-STAT3 / STAT3，Bax，TGF-β1， FN，collagenIV and α-SMA decreased ( P ＜0. 05 ) ; inhibition of miR-30a-5p attenuated the promoting effect of knockdown of circ_WBSCR17 on proliferation of HMCL cells，and enhanced apoptosis，cellular fibrosis and inflammatory responses ; FISH experiment confirmed that WBSCR17 was mainly distributed in the cytoplasm ; dual-luciferase reporter gene experiment confirmed that circ_WBSCR17，JAK1 and miR-30a-5p had a targeted regulatory rela- tionship．@*Conclusion @#Knockdown of circ_WBSCR17 can reduce high glucose-induced fibrosis and inflammation in human mesangial cells by regulating the miR-30a-5p /JAK1 axis.