Study on the effects and mechanism of Liqi Huoxue Drop Pill-containing serum on ISO-induced transdif-ferentiation of suckling rat cardiac fibroblasts
OBJECTIVE To investigate the effect of Liqi Huoxue Drop Pill-containing serum on isoprenaline(ISO)induced transdifferentiation of suckling rat cardiac fibroblasts(CFs)and its mechanism.METHODS In vitro culture of SD suckling mouse cardiac fibroblasts,and the CCK-8 method was used to screen the optimal working concentration of Liqi Huoxue Drop Pilll-containing serum.The cells were divided into normal control group(NC group),blank rat serum group(BRS group),ISO induced model group(NC+ISO group),blank rat serum+ISO induced model group(BRS+ISO group),Liqi Huoxue Drop Pill-containing serum+ISO induced model group(LQHXDP+ISO group).The levels of hydroxyproline(Hyp)and TGF-β1 were detected by biochemical assay and ELISA,respectively,and the cell cycle distribution and protein expression of CollagenⅠ,Collagen Ⅲ,α-SMA,IL-6,p-STAT3 and STAT3 were detected by flow cytometry and Western blot,respectively.RESULTS The cells identified by Vimentin immunofluorescence as CFs,and the drug-containing serum prepared by 10％Liqi Huoxue Drop Pill at a dose of 175.0 mg·kg-1 was the optimal working concentration.Compared with the NC and BRS groups,the proliferation viability of ISO-induced CFs was significantly increased,the contents of Hyp and TGF-β1 in the cell culture supernatant were significantly increased,the number of S phase cells was significantly increased,and the expression of CollagenⅠ,Collagen Ⅲ,α-SMA,IL-6 and p-STAT3 proteins was significantly increased.Compared with the BRS+ISO group,the proliferation activity of CFs in the LQHXDP+ISO group decreased,the content of Hyp and TGF-β1 was significantly reduced,the S phase cells were significantly reduced,and the expression of Collagen Ⅰ,Collagen Ⅲ,α-SMA,IL-6 and p-STAT3 pro-teins were significantly reduced.CONCLUSION Liqi Huoxue Drop Pill-containing serum can significantly inhibit the transdiffer-entiation of ISO-induced CFs,which is related to its inhibition of ISO-induced proliferation and activity of CFs,reduce the secre-tion of Hyp and TGF-β1,reduce collagen production and inhibit IL-6/STAT3 signaling pathway.
Liqi Huoxue Drop Pillmyocardial fibrosiscardiac fibroblastsIL-6/STAT3 signaling pathway
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