雷公藤多苷对强直性脊柱炎患者成纤维细胞中BMP-2表达的影响

摘要

目的：探讨雷公藤多苷（LGTDG）对骨形态发生蛋白（BMP）信号转导通路及 BMP-2表达的影响，阐明 LGTDG抗强直性脊柱炎（AS）骨化的作用机制。方法：体外培养的 AS成纤维细胞分为对照组与0.5、1.0、1.5和2.0 mg · L－1给药组，检测各组细胞内碱性磷酸酶（ALP ）活性及最佳药物浓度；CCK-8法检测1.0 mg· L－1给药组的细胞增殖率；生化检测法定量检测对照组与1.0 mg · L－1给药组 BMP-2表达水平；Western blotting法检测对照组与1.0 mg·L－1给药组BMP-2和Cbfal蛋白表达。结果：各给药组细胞中ALP活性均低于对照组，其中以1.0 mg·L－1给药组细胞的ALP活性为最低（P＜0.01）。CCK-8法检测，1.0 mg·L－1给药组 AS成纤维细胞增殖率明显低于对照组（P＜0.01）， LDTDG诱导第4天细胞增殖率最高，进入平台期后细胞的增殖率开始下降。生化检测法和 Western blotting法检测，1.0 mg·L－1给药组BMP-2的蛋白表达明显低于对照组（P＜0.01）。结论：LGTDG通过对BMP信号转导通路的影响有效地抑制 AS成纤维细胞内 BMP-2表达，延缓细胞向成骨型分化导致 AS骨化发生。%Objective To study the influence of tripterygium glycosides (LGTDG)in the bone morphogenetic protein(BMP)signal transduction pathway and BMP-2 expression,and to clarify the mechanism of anti-ankylosing spondylitis (AS)ossification of LGTDG.Methods The in vitro cultured AS fibroblasts were divided into control group and 0.5,1.0,1.5,2.0 mg·L-1 LGTDG groups.The alkaline phosphatase (ALP)activities of the cells and optimal drug concentrations in various groups were detected;CCK-8 assay was used to detect the proliferation rate of the cells in 1.0 mg· L-1 LGTDG group;the biochemical tests were performed to quantitatively detect the BMP-2 expression levels in control group and 1.0 mg· L-1 LGTDG group;Western blotting method was used to determine the BMP-2 and Cbfal protein expressions. Results The ALP activities in LGTDG groups were lower than that in control group,especially in 1.0 mg·L-1 LGTDG group (P<0.01).The CCK-8 assay results showed that the proliferation rate of AS fibroblasts in 1.0 mg·L-1 LGTDG group was significantly low than that in control group(P<0.01),the proliferation rate reached the peak at the 4th day after LGTDG treatment and entered into the plateau phase,then the proliferation rate of the cells was decreased.The biochemical assay and Western blotting results indicated that the protein expression levels of BMP-2 in 1.0 mg·L-1 LGTDG group was significantly lower than that in control group (P<0.01).Conclusion LGTDG can effectively inhibit the BMP-2 expression in AS fibroblasts and delay the cells to differentiate into the osteoblasts and lead to AS ossification by BMP signal transduction pathway.