Effects of Berberine Hydrochloride on Fibroblast-Like Synovial Cells of Ankylosing Spondylitis
Objective:To investigate effect and molecular mechanisms of berberine hydrochloride on the proliferation,migration,and invasion of fibroblast-like synovial(FLS)cells in lipopolysaccharide induced ankylosing spondylitis.Meth-ods:FLS cells were isolated from synovial tissue biopsy of patients with ankylosing spondylitis and cultured.After the FLS cells were transferred to the third generation,the cells were pre-treated with lipopolysaccharide for 24 h,and then dried with culture media containing different concentrations of berberine hydrochloride for 24 h.Subsequently,an inverted mi-croscope was used to observe the morphological changes of berberine hydrochloride on FLS cells.CCK-8 was used to de-tect the effect of berberine hydrochloride on the proliferation of FLS cells under lipopolysaccharide stimulation.Plate clone formation experiments and Calcein/PI staining experiments were used to examine the effect of berberine hydrochloride on the proliferation of FLS cells under lipopolysaccharide stimu-lation;Hoechst33342 staining experiment and Annexin-V/PI flow cytometry were used to detect the effect of berberine hydrochloride on apoptosis of FLS cells stimulated by lipopolysaccharide.The effects of berberine hydrochloride on the ex-pression levels of IL-6,TNF-α,IL-17 and IL-23 in the supernatant of FLS by lipopolysaccharide were detected by ELISA.Transwell chamber experiment was used to detect the effect of berberine hydrochloride on the invasive ability of FLS cells;scratch experiment was used to detect the effect of berberine hydrochloride on the migration ability of FLS cells,and protein blotting technology was used to detect the effect of berberine hydrochloride on apoptosis,migration,and pathway protein expression levels in FLS cells under lipopolysaccharide stimulation.Results:Compared with the blank group,the berberine hydrochloride treatment group had a certain inhibitory effect on the proliferation,invasion,migration,and inflam-matory factors of FLS cells under lipopolysaccharide stimulation,and induced apoptosis of FLS cells under lipopolysaccha-ride stimulation.Berberine hydrochloride can up-regulate the relative expression of Bax,cleared casease-3,and E-cadherin(P＜0.01),and down-regulated the expression levels of Bcl-2,IL-17,IL-23,c-Jun NH2 terminal kinase,extracellular sig-nal-regulated protein kinase,and N-cadherin(P＜0.01).Conclusion:BBR inhibits the proliferation,invasion,and migration ability of AS FLS cells and induces their apoptosis,which may be related to the inhibition of the p-JNK and p-ERK1/2 pathways.
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