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目的探讨Sonic Hedgehog(SHH)在滑膜成纤维细胞增殖中的作用。方法收集类风湿性关节炎(RA)、系统性红斑狼疮(SLE)、强直性脊柱炎(AS)患者及健康正常人血清样本各(30例),ELISA检测上述血清SHH的含量。SD大鼠皮内注射2型胶原蛋白(Col2)诱导RA大鼠模型(CIA),取其滑膜组织原代培养滑膜成纤维细胞(SF)。免疫荧光细胞化学染色法检测vimentin表达鉴定SF,并检测SHH在SF中的表达。培养SF给予SHH-胶质瘤相关癌基因1(Gli-1)通路特异性阻断剂GANT61处理72 h,Western blot法检测SF表达SHH的变化情况,CCK-8法检测SF的增殖情况。结果 RA患者血清中SHH的含量较SLE、AS患者及正常组含量升高。成功建立CIA模型及分离培养SF;CIA-SF表达SHH较正常组SF高。给予GANT61处理72 h,CIA-SF中SHH蛋白表达降低且细胞增殖水平下降。结论 SHH参与RA发病与CIA-SF增殖有关。
Objective To investigate the role of Sonic Hedgehog (SHH) in the proliferation of synovial fibroblasts. Methods Serum samples from patients with rheumatoid arthritis (RA), systemic lupus erythematosus (SLE), ankylosing spondylitis (AS) and normal healthy subjects were collected, respectively. The levels of serum SHH were measured by ELISA. SD rats were injected intradermally with type 2 collagen (Col2) to induce the RA rat model (CIA), and the primary synovial fibroblasts (SF) were cultured in vitro. The expression of vimentin was detected by immunofluorescence cytochemical staining to identify SF, and the expression of SHH in SF was detected. SF was treated with GliCl, a specific inhibitor of glioma-associated oncogene 1 (Gli-1), for 72 h. Western blot was used to detect SHH expression in SF. CCK-8 assay was used to detect the proliferation of SF. Results The serum levels of SHH in RA patients were higher than those in SLE, AS patients and normal controls. CIA model was successfully established and SF was isolated and cultured. SHH of CIA-SF expression was higher than that of normal group. After GANT61 treatment for 72 h, the expression of SHH protein in CIA-SF decreased and the cell proliferation decreased. Conclusion The involvement of SHH in the pathogenesis of RA is related to the proliferation of CIA-SF.