Effects of berberine hydrochloride on proliferation and osteogenic differentiation of fibroblast-like synoviocytes derived from patients with ankylosing spondylitis in vitro
Objective:To investigate the effects of berberine hydrochloride(BBH)on proliferation and osteogenic differentiation of fi-broblast-like synoviocytes(FLSs)derived from patients with ankylosing spondylitis(AS)in vitro.Methods:The FLSs were isolated from the hip synovial tissues of AS patients and cultured,and then identified by immunofluorescence staining for subsequent experiments.The third-generation FLSs were collected and pretreated with interleukin(IL)-17 and IL-23,followed by intervention with BBH at concentrations of 0(blank group),80(low-dose BBH group),120(medium-dose BBH group)and 200 μmol/L(high-dose BBH group),respectively.After the end of intervention,the cellular proliferation was observed by using optical microscopy,and the levels of inflammatory factors and bone ho-meostasis regulatory factors were detected by using enzyme-linked immunosorbent assay(ELISA),meanwhile,the cellular osteogenic differ-entiation was observed via alizarin red staining(ARS).Results:① The levels of IL-17,IL-23,IL-6 and tumor necrosis factor-α(TNF-α)were lower in low-,medium-,and high-dose BBH group compared to blank group(IL-17:P=0.046,P=0.005,P=0.004;IL-23:P=0.024,P=0.002,P=0.000;IL-6:P=0.012,P=0.000,P=0.000;TNF-α:P=0.012,P=0.004,P=0.001),and were lower in medi-um-and high-dose BBH group compared to low-dose BBH group(IL-17:P=0.034,P=0.024;IL-23:P=0.020,P=0.000;IL-6:P=0.000,P=0.000;TNF-α:P=0.012,P=0.002),and were lower in high-dose BBH group compared to medium-dose BBH group(IL-17:P=0.029;IL-23:P=0.014;IL-6:P=0.005;TNF-α:P=0.026).②The results of observation by optical microscopy showed that,com-pared to blank group,the intercellular space and the number of apoptotic cells increased in low-,medium-,and high-dose BBH group,with a dose-dependence exhibited in the degree of change.③Compared with that of blank group,the ARS intensity was reduced in low-,medium-,and high-dose BBH group,with a dose-dependence exhibited in the degree of reduction.④The levels of Dickkopf-related protein 1(DKK1)and receptor activator of NF-κB ligand(RANKL)were lower in low-,medium-,and high-dose BBH group compared to blank group(DKK1:P=0.048,P=0.032,P=0.001;RANKL:P=0.046,P=0.021,P=0.001),and were lower in medium-and high-dose BBH group com-pared to low-dose BBH group(DKK1:P=0.028,P=0.002;RANKL:P=0.047,P=0.002),and were lower in high-dose BBH group compared to medium-dose BBH group(DKK1:P=0.018;RANKL:P=0.011).Conclusion:BBH can inhibit the proliferation of FLSs de-rived from AS patients in vitro,and repress the osteogenic differentiation of FLSs via reducing the bone homeostasis regulatory factors RANKL and DKK1 secreted by FLSs,with a dose-dependence exhibited in the effects.
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