IHH-Gli信号通路在CIA大鼠滑膜成纤维细胞增殖中的作用

Roles of IHH-Gli signaling in synovial fibroblasts proliferation in CIA

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。

This study performed to investigate whether Indian Hedgehog-Gli（IHH-Gli） signaling played a role in the proliferation of SF in collagen induced arthritis（CIA）. The CIA rats were induced by injecting the collagen,and confirmed by pathology and imaging examination. The knee synovial tissues of the CIA rats were cultured to obtain the synovial fibroblast（SF）, and the purity of the SF was about 95% confirmed by detecting the Vimentin.The IHH-Gli signaling related components（IHH, Ptc, Smo and Gli1） were expressed in CIA-SF examined by immunofluorescence, and the IHH of CIA-SF was higher than that of normal SF. GANT61, a specific inhibitor of HH-Gli signaling, significantly decreased CIA-SF proliferation at 10 μmol/L and 15 μmol/L and decreased the expression of GSK-3β in CIA-SF. Our data show that IHH-Gli signaling plays a role in the proliferation of SF in CIA; IHH-Gli signaling may therefore maybe a novel therapeutic target in RA treatment.