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目的初步探讨IHH-Gli信号通路在胶原诱导型关节炎(collagen induced arthritis,CIA)大鼠滑膜成纤维细胞(synovial fibroblast,SF)增殖中的作用。方法注射胶原诱导CIA大鼠模型,采用组织块贴壁法培养大鼠膝关节SF,免疫荧光法检测Vimentin鉴定细胞纯度,免疫荧光法检测IHH-Gli通路相关分子(IHH、Ptc、Smo及Gli1)的表达情况,给予GANT61阻断IHH-Gli信号通路,观察细胞形态变化、CCK8法检测细胞增殖情况、免疫荧光法检测糖原合成酶激酶-3β(glycogen synthase kinase-3β,GSK-3β)表达变化情况。结果病理及影像学检查显示CIA模型建立成功,培养的细胞95%以上均表达Vimentin,CIA-SF表达IHH、Ptc、Smo及Gli1蛋白,且CIA-SF表达IHH蛋白较正常SF高,10μmol/L及15μmol/L的GANT61能显著抑制CIA-SF的增殖,且降低GSK-3β的蛋白表达水平。结论 IHH-Gli信号通路参与CIA-SF的增殖机制,可能成为研究RA治疗药物的新靶点。
Objective To investigate the role of IHH-Gli signaling pathway in the proliferation of synovial fibroblasts (SF) induced by collagen induced arthritis (CIA) in rats. Methods CIA rat model was induced by collagen injection. The knee joint SF was cultured by tissue adhesion method. The purity of cells was detected by immunofluorescence and IHH-Gli pathway related molecules (IHH, Ptc, Smo and Gli1) GANT61 was used to block the IHH-Gli signaling pathway, the morphological changes of cells were observed, the cell proliferation was detected by CCK8 assay, and the expression of glycogen synthase kinase-3β (GSK-3β) was detected by immunofluorescence staining Happening. Results Pathology and imaging examination showed that CIA model was successfully established. Vimentin was expressed in more than 95% of cells, IHH, Ptc, Smo and Gli1 proteins were expressed in CIA-SF, and IHH protein in CIA-SF was higher than that in normal SF, 10μmol / L And 15μmol / L GANT61 could significantly inhibit the proliferation of CIA-SF and decrease the protein expression of GSK-3β. Conclusion The IHH-Gli signaling pathway is involved in the proliferation mechanism of CIA-SF, which may be a new target for the study of RA therapeutic drugs.