摘要
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 effect of sonic hedgehog （SHH） on the proliferation of synovial fibroblasts （SFs）. Methods The serum samples were collected from 30 rheumatoid arthritis （RA） patients, 30 systemic lupus erythematosus （SLE） patients, 30 ankylosing spondylitis （AS） patients and 30 healthy subjects. The concentrations of serum SHH were detected by ELISA. Collagen induced arthritis （CIA） were developed by type 2 collagen in Sprague-Dawley rats. The SFs were isolated from knee synovial tissues of CIA rats, and then identified by the detection of vimentin by immunofluorescence technique. Before and 72 hours after blocking SHH-glioma-associated oncogene 1 （ Gli-1 ） signaling pathway with GANT61, the expression level of SHH in SFs was detected by Western blotting, and the proliferation of SFs was examined with CCK-8 assay. Results The level of serum SHH in the RA patients was remarkably higher than that in the SLE, AS patients and the healthy controls. In the CIA rats, the expression of SHH in SFs in vitrowas higher than that in the healthy control rats. After 72-hour treatment of GANT61 to block SHH-Gli-1 signaling pathway, the expression level of SHH protein in SFs from CIA rats was reduced, and meanwhile the proliferation of the SFs was inhibited. Conclusion SHH plays an important role in the proliferation of SFs and could be used as a potential therapeutic target for RA.
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2016年第5期630-634,共5页
Chinese Journal of Cellular and Molecular Immunology
基金
中国博士后基金(2015M581864)
江苏省博士后科研资助计划A(1501061)
江苏省教育厅高校自然科学研究基金(15KJB310025)
江苏省脑病生物信息重点实验室开放课题(1507)
江苏高校优势学科建设工程资助项目(PAPD)
