Cyclopamine在佐剂性关节炎大鼠肾脏损伤中的保护作用及机制研究

目的：观察佐剂性关节炎(AA) 大鼠Hedgehog信号通路活化，并探讨Cyclopamine对大鼠关节炎症及肾脏损伤的影响。方法：将40只大鼠随机分为空白组、Cyclopamine组、AA+Cyclopamine组、AA模型组。采用弗氏完全佐剂建立AA大鼠模型，使用Cyclopamine腹腔注射，并通过测量足爪肿胀、全身炎症反应及关节炎症评分的方法进行半定量评价。HE染色检测各组大鼠肾脏病理改变，Western blot检测各组大鼠肾脏Gli1蛋白表达水平，免疫组织化学法染色检测各组大鼠肾脏TNF-α、IFN-γ、IL-6表达。结果：使用Cyclopamine后，能够明显降低AA大鼠足爪肿胀度和改善AA大鼠的关节炎指。与对照组相比，AA模型组大鼠Cr、BUN和脏器系数出现明显升高改变，差异有统计学意义，同时肾脏电镜病理检测发现AA模型组大鼠出现明显的病理改变，AA大鼠使用抑制剂Cyclopamine后能够明显降低肌酐（Cr）、尿素氮（BUN）和脏器系数改变。Western blot检测各组肾脏组织中Gli1的蛋白表达，发现对照组与Cyclopamine组相比Gli1蛋白表达无明显差异，AA模型组及AA+Cyclopamine组Gli1蛋白表达量明显高于对照组，差异有统计学意义，AA+Cyclopamine组与AA模型组相比，Gli1蛋白表达水平明显有下降趋势，差异有统计学意义；免疫组化法检测肾脏组织中促炎因子TNF-α、IFN-γ、IL-6表达改变情况并进行半定量评分，与空白组相比，AA模型组及AA+Cyclopamine组肾脏TN-α、IFN-γ表达明显升高，且使用Cyclopamine后，AA大鼠肾脏TNF-α、IFN-γ表达显著降低，AA模型组肾脏IL-6表达较对照组明显升高。结论：Cyclopamine能够明显改善AA大鼠的关节炎症和肾脏损伤程度，在此过程中Hh通路处于活化状态，并诱导炎性因子表达改变。

Protective effects of Cyclopamine in adjuvant arthritis rats kidney injury and mechanism research

Objective:Observed activity of adjuvant arthritis (AA) rats Hedgehog signaling pathway and explore the effect of Cyclopamine on rat arthritis and kidney injury.Methods: 40 rats were randomly divided into control group,Cyclopamine group,AA+Cyclopamine group and AA model group.We used Freund′s complete adjuvant rat model with Cyclopamine intraperitoneal injection.By measuring paw swelling,systemic inflammation and arthritis semi-quantitative assessment methods of rats to evaluate the model.HE staining was used to detect the renal pathological changes of rats.Western blot was used to detect kidney Gli1 protein expression levels of the rats.Immunohistochemical staining was used to detect the rat kidney TNF-α,IFN-γ,IL-6 expression.Results: After using Cyclopamine,the AA rat paw swelling reduced and arthritis refers relieved significantly.Compared with the control group,Cr,BUN and organ coefficient increased significantly in AA model group,and the difference was statistically significant.Meanwhile renal TEM detection appeared obvious pathological changes in rats of AA model groupAfter using cyclopamine,the content of Cr,BUN and organ coefficient change reduced significantly,so did the pathological.Western blot detected kidney tissue Gli1 protein in each group.Compared to control group,there was no significant difference in Cyclopamine group.AA model group and AA+Cyclopamine group Gli1 protein expression was significantly higher compared with Cyclopamine Gli1 protein group,the difference was statistically significant.AA model group and AA+ Cyclopamine group Gli1 protein expression was significantly higher,the difference was statistically significant.Compared to the AA model group,AA+Cyclopamine group Gli1 protein levels have decreased significantly,the difference was statistically significant.Immunohistochemical assay detection found that kidney tissue proinflammatory cytokines TNF-α,IFN-γ,IL-6 expression and semiquantitative score changed.Compared with the control group,TNF-α,IFN-γ expression of AA model group were significantly increased.After using Cyclopamine,the expression of TNF-α,IFN-γ in kidney of AA model reduced significantly.IL-6 expression in AA model group was significantly higher than the control group.Conclusion: Cyclopamine AA can relieve arthritis and kidney injury in rats with arthritis AA,the Hh pathway was on activity state in the process,may altered expression of inflammatory factors.