穿山龙总皂苷对胶原诱导性关节炎大鼠关节滑膜组织NF-κB p65活性及STAT3表达的影响

Effects of Total Saponin from Rhizoma Dioscreae Nipponicae on NF-κB p65 Activity and STAT3 Protein Expression in Joint Synovium of CIA Rats

目的通过观察穿山龙总皂苷对胶原诱导性关节炎(CIA)大鼠滑膜组织核转录因子κB(NF-κB)及信号转导子和转录激活因子3(STAT3)表达的影响,探讨穿山龙总皂苷抑制类风湿性关节炎血管新生可能的信号转导通路作用机制。方法将造模成功的60只CIA大鼠随机分为CIA模型组、雷公藤组(阳性对照组)、穿山龙总皂苷组、薯蓣皂苷元组、穿山龙水溶性皂苷片组,另设正常对照组,每组12只。造模成功后分别给予相应的药物灌胃,连续治疗35 d。应用TransAMTMNF-κB p65活性检测试剂盒检测滑膜组织中NF-κB p65的DNA结合活性。应用免疫组织化学染色的方法观察滑膜组织中STAT3蛋白的表达。结果与正常对照组相比,CIA模型组大鼠滑膜细胞核蛋白提取物中NF-κB p65的DNA结合活性和滑膜组织中STAT3蛋白的表达显著增高,有统计学意义(P<0.01);与CIA模型组相比,穿山龙总皂苷组、雷公藤组、薯蓣皂苷元组、穿山龙水溶性皂苷片组的NF-κB p65的DNA结合活性和STAT3蛋白的表达均显著降低,有统计学意义(P<0.01);各治疗组之间两两比较均无统计学意义(P>0.05)。结论穿山龙总皂苷可能通过抑制NF-κB信号转导通路中NF-κB p65的活性和JAK-STAT信号转导通路中重要底物STAT3蛋白的表达,起到抑制类风湿性关节炎血管新生的作用。

Objective To explore the effects of total saponin from rhizoma diosereae nipponicae （RDN） on NF-κB p65 activity and STAT3 expression in synovium of collagen-induced arfitis （CIA） rats, and investigate the underlying mechanism about signal transduction pathways of RI）N on antiangiogenesis in treating rheumatoid arthritis. Methods A total of 60 rats with successfttl modeling were randomly divided into 5 groups （n = 12 ） ： CIA model group, Tripterygium （TP）-treating group （ positive control group ）, RDN-treating group, Diosgenin-treating group and Diosgenin-Tablets-treating group,and established a separate normal control group （n = 12）,then the rats in treated groups were lavaged with corresponding medicine for 35 days. The NF-κB p65 activity was detected by TransAMIM NF-κB p65 kit. Immunohistochemical staining was employed to determine the expression of STAT3 protein in synovimn. Results NF-KB p65 activity and the expression of STAT3 protein were more obviously increased in synovium of CIA model group than in normal control group （P 〈 0.01 ）. The NF-KB p65 activity and the expression of STAT3 protein in RDN-treating, TP-treating, Diosgenin-treating and Diosgenin-tablet-treating groups were more obviously decreased than in CIA model group （P 〈 0.01）. There was no significant difference between each treated group （P 〉 0.05）. Conclusion Total saponin from RDA can inhibit angiogenesis in synovial of RA,which may be related to the inhibition of NF-κB p65 activity in NF-κB signaling pathway and the down-regulated expression of STAT3 protein which is an important substrate in the JAK-STAT signaling pathway.