补肾通督胶囊对胶原诱导关节炎大鼠RANK/RANKL/OPG系统的影响

Effect of Bushen Tongdu Capsule on RANK / RANKL / OPG Pathway of Collagen Induced Arthritis Rats

目的研究补肾通督胶囊对胶原诱导关节炎大鼠RANK/RANKL/OPG系统的影响,为中医药治疗类风湿关节炎(RA)提供理论依据。方法 RA动物模型选用胶原诱导关节炎模型,将大鼠分为正常对照组、模型组、补肾通督胶囊低剂量组(BSL)、中剂量组(BSM)、高剂量组(BSH)和雷公藤组6组,每组7只大鼠。造模后13天开始给药,BSL、BSM及BSH组分别给予120、240及480 mg/(kg·d)的补肾通督胶囊灌胃,雷公藤组给予雷公藤多苷片24 mg/(kg·d)灌胃。各组每天灌胃1次,每次2 m L。正常对照组和模型组给予2 m L的生理盐水灌胃。共灌胃18天。于31天取材,制作踝关节TRAP切片,采用ELISA法检测血清核因子κB受体活化因子配体(RANKL)、骨保护素(OPG)、OPG巨噬细胞集落刺激因子(M-CSF)。结果与正常对照组比较,模型组大鼠踝关节病理切片的阳性反应、炎症和骨破坏程度显著提高(P<0.01),血清RANKL和M-CSF水平皆有显著上升(P<0.01),而OPG和OPG/RANKL水平皆有显著降低(P<0.01)。与模型组比较,BSH组和雷公藤组阳性反应、炎症和骨破坏评分也显著下降(P<0.01);而BS各组和雷公藤组RANKL和M-CSF均显著下调,而OPG和OPG/RANKL均显著上调,差异有统计学意义(P<0.01)。与雷公藤组比较,正常组M-CSF较低,而OPG和OPG/RANKL均较高,差异有统计学意义(P<0.01),而模型组和BS各组RANKL和M-CSF均显著上调,而OPG和OPG/RANKL均显著下调(P<0.01)。结论补肾通督胶囊对于关节炎大鼠缓解骨破坏的机制,可能是通过对破骨细胞的调控起作用。

Objective To study the effect of Bushen Tongdu Capsule( BTC) on RANK / RANKL / OPG pathway of collagen induced arthritis( CIA) rats,thereby laying theoretic evidence for treating rheumatic arthritis( RA) by Chinese medicine.Methods RA model was induced by CIA. Totally 42 rats were randomly divided into six groups,i. e.,the normal control group,the model group,the low dose BTC( BSL) group,the medium dose BTC( BSM) group,the high dose BTC( BSH)group,and the Tripterygium Glycosides( TG) group,7 in each group. BTC at the daily dose of 120,240,and 480 mg / kg was given by gastrogavage to rats in the BSL,BSM,and BSH group respectively from the 13 th day of modeling. TG at the daily dose of 24 mg / kg was given by gastrogavage to rats in the TG group. All medication was given once daily,2 m L each time. Two m L normal saline was administered to rats in the normal control group and the model group. All medication lasted for 18 days. Sam-ples were taken at day 31. The TRAP section of the ankle joint was fixed in 10% formalin for TRAP stain. Serum levels of osteoprotegerin( OPG),receptor activator of nuclear factor- k B ligand( RANKL),and macrophage colony- stimulating factor( M- CSF) were detected using ELISA. Results Compared with the normal control group,positive reactions of pathological ankle joint section,inflammation,and osteoclasia degree were significantly improved in the model group,serum levels of RANKL and M- CSF were up- regulated,levels of OPG and OPG / RANKL were significantly lowered( all P < 0. 01). Compared with the model group,positive reactions of pathological ankle joint section,inflammation,and osteoclasia degree also significantly decreased in the BSH group and the TG group( all P < 0. 01). RANKL and M- CSF were significantly down- regulated in each medicated group,while levels of OPG and OPG / RANKL were significantly up- regulated( all P < 0. 01). Compared with the TG group,M- CSF was lower,but levels of OPG and OPG / RANKL were significantly up- regulated in the normal control group( all P < 0. 01). RANKL and M- CSF were significantly up- regulated,while levels of OPG and OPG /RANKL were significantly down- regulated in the model group and each BS group( all P < 0. 01). Conclusion BTC could relieve bone damage of CIA rats possibly through regulating and controlling osteoclasts.