[Effects of Huatan Tongluo Recipe on IL-1β-induced Proliferation of Rheumatoid Arthritis Synovial Fibroblasts and the Production of TNF-α and aFGF].

Abstract

Objective To observe the effects of Huatan Tongluo Recipe (HTTLR) on the proliferation of IL-β p induced rheumatoid arthritis synovial fibroblast ( RASFB) and secretion of necrosis factor α (TNF-α) and acidic fibroblast growth factor (aFGF) in vitro. Methods RASFB cell line was cultured in vitro and stimulated by IL-1β. The proliferation of RASFB was detected using WST-1 after adding IL-1β with final concentrations of 1 , 5, 10, 20 μg/L for 24 and 48 h respectively. Then 20 μg/L IL-1β recruited as induction dose was set up as IL-1β group. High, middle, low dose HTTLR groups were set up by adding HT- TLR decoction with final concentration of 5%, 2%, 1% (V/V) , respectively for 24 and 48 h. A blank con- trol group was also set up. The proliferation rates were compared. Contents of TNF-α and aFGF were detected in each group using ELISA. mRNA expressions of TNF-α and aFGF were detected using RT-PCR. Results The proliferation rates of RASFB at 24 h and 48 h were lower at 1 μg/L IL-1 β than at 5, 10, 20 μg/L IL-1β (P <0. 01). The proliferation rate of RASFB was higher at 10 and 20 μg/L IL-1β than at 5 μg/L IL-1β (P <0. 01). Besides, the proliferation rate of RASFB was higher at 20 μg/L IL-1β than at 10 μg/L IL-1 β (P <0. 01). The proliferation rate of RASFB was higher at 48 h than at 24 h (P <0. 01). Com- pared with the high dose HTTLR group, the proliferation rate of RASFB was lowered in middle and low dose HTTLR groups (P <0. 01). Besides, IL-1β induced proliferation rate of RASFB was obviously reduced in the middle dose HTTLR group (P <0. 01). Compared with the blank control group, mRNA ex- pressions of TNF-α and aFGF and their contents were elevated in the IL-1β group at 24 and 48 h (P < 0. 05). Compared with the IL-1 β group, mRNA expressions of TNF-α- and aFGF and their contents, except TNF-α- mRNA expression in the low dose HTTLR group at 24 h, were all obviously lowered in 3 dose HTTLR groups at 24 h and 48 h (P <0. 05). Compared with the high dose HTTLR group, mRNA expressions of TNF-(α and aFGF increased in middle and low dose HTTLR groups at 24 h and 48 h; TNF-α content in the low dose HTTLR group at 24 h; contents of TNF-α and aFGF in middle and low dose HTTLR groups at 24 h and 48 h all increased (P <0. 05). Conclusion The mechanism of HTTLR treatment for RA might be related to inhibiting RASFB proliferation, and decreasing mRNA expressions of TNF-α and aFGF as well as their protein secretion.