肾福康胶囊通过STAT3/YAP通路调控细胞串扰改善肾间质纤维化的机制OA

OBJECTIVE To investigate the mechanism by which Shenfukang capsule(SFK)ameliorates renal interstitial fibrosis(RIF)by regulating macrophage-fibroblast crosstalk via the signal transducer and activator of transcription 3(STAT3)/Yes-associated protein(YAP)pathway.METHODS RAW264.7 macrophages were induced to polarize with transforming growth factor-β1(TGF-β1).Subsequently,a non-contact co-culture system of macrophages with renal fibroblasts NRK-49F,as well as a non-contact co-culture system of macrophages with YAP-knockdown renal fibroblasts,were established.Cells were treated with low,medium,and high concentrations(4,8,16 μg/mL)of SFK,as well as a STAT3 inhibitor(STAT3-I,1.64 μg/mL)and losartan potassium tablets(positive control,0.28 μg/mL),for 48 h.After intervention,the protein and mRNA expression levels of CD86,CD163,and STAT3 in macrophages,as well as CD86,F4/80,and STAT3 mRNA,were detected.In co-cultured renal fibroblasts,the protein and mRNA expression levels of α-smooth muscle actin(α-SMA),Vimentin,collagen type Ⅰ(Col-Ⅰ),matrix metalloproteinase-1(MMP-1),and YAP were detected.In co-cultured YAP-knockdown renal fibroblasts,the protein and mRNA expression levels of α-SMA,Vimentin,and MMP-1 were also detected.RESULTS Following TGF-β1 induction,the protein expression levels of CD86,CD163,and STAT3,as well as the mRNA expression levels of CD86,F4/80,and STAT3 in macrophages were significantly increased(P＜0.05).The conditioned medium from polarized macrophages activated renal fibroblasts,as evidenced by significantly increased protein and mRNA expression levels of α-SMA,Vimentin,Col-Ⅰ,and YAP,and significantly decreased protein and mRNA expression levels of MMP-1 in renal fibroblasts(P＜0.05).Treatment with SFK and STAT3-I reversed the changes in the above indicators,with the medium concentration SFK group showing stronger effects on some indicators than the low and high concentration groups.In the non-contact co-culture experiment of macrophages and YAP-knockdown renal fibroblasts,there were no statistically significant differences in the protein and mRNA expression of α-SMA,Vimentin,and MMP-1 among the groups.CONCLUSIONS SFK can inhibit macrophage-renal fibroblast crosstalk by blocking the STAT3/YAP pathway,thereby delaying the progression of RIF.