骨灵膏及其拆方对骨关节炎模型大鼠炎性细胞因子与聚蛋白多糖酶的影响

Effect of Gulinggao(骨灵膏)and Its Disassembled Prescriptions on Inflammatory Cytokines and Aggrecanase of Osteoarthritis Model Rats

目的探讨骨灵膏及其拆方制剂抗骨关节炎软骨损伤的可能作用机制。方法 60只SD大鼠随机分为骨灵膏组、骨膏组、灵膏组、塞来昔布组、模型组及正常组,每组10只。除正常组外其余各组采用寒冷刺激结合石膏固定的方法建立骨关节炎模型,造模成功后骨灵膏组、骨膏组及灵膏组分别给予骨灵膏、骨膏及灵膏生药量33.31g/(kg·d)灌胃给药,塞来昔布组给予塞来昔布20.83 mg/(kg·d)灌胃,模型组和正常组给予等量生理盐水灌胃,8周后检测膝关节软骨组织聚蛋白多糖酶4(ADAMTs-4)、聚蛋白多糖酶5(ADAMTs-5)蛋白表达,检测膝关节滑膜和血清白细胞介素1β(IL-1β)、肿瘤坏死因子α(TNF-α)、前列腺素E-2(PGE_2)、环氧合酶2(COX-2)、γ干扰素(IFN-γ)的含量。结果与正常组比较,模型组大鼠血清与滑膜IL-1β、TNF-α、PGE_2、COX-2、IFN-γ的含量均显著增加(P<0.01)。与模型组比较,骨灵膏组血清与滑膜IL-1β、TNF-α、PGE_2、COX-2、IFN-γ含量明显降低(P<0.01)。与骨灵膏组比较,骨膏组血清IL-1β、TNF-α、COX-2、IFN-γ含量和滑膜IL-1β、COX-2、PGE_2、TNF-α、IFN-γ含量升高(P<0.05或P<0.01),灵膏组血清和滑膜IL-1β、TNF-α、COX-2、PGE_2、IFN-γ含量升高(P<0.05或P<0.01),塞来昔布组血清IL-1β、TNF-α、PGE_2、IFN-γ含量和滑膜IL-1β、PGE_2含量亦升高(P<0.01)。各组大鼠膝关节软骨组织ADAMTs-4、ADAMTs-5均有表达,其中模型组表达最显著,骨灵膏组ADAMTs-4、ADAMTs-5蛋白表达最低。结论骨灵膏可能通过降低血清及滑膜中炎性细胞因子,进而阻止局部关节软骨聚蛋白多糖酶合成,从而减轻了骨关节炎软骨损伤,并且作用明显优于其拆方制剂及塞来昔布。

Objective To explore the possible mechanism of Gulinggao（骨灵膏） and its disassembled prescriptions in resisting cartilage injury in osteoarthritis.Methods Randomly divided 60 SD rats into the Gulinggao group,the Gugao（骨膏） group,the Linggao（灵膏） group,the Celecoxib group,the model group and the normal group,with 10 each.All groups except the normal group adopted cold stimulating with plaster fixation method to establish osteoarthritis model.After modeling successfully,the Gulinggao group,the Gugao group and the Linggao group were respectively given Gulinggao,Gugao and Linggao of crude herb 33.31 g/（kg· d） for gavage.The Celecoxib group was given celecoxib 20.83 mg/（kg · d） for gavage.The model group and the normal group was given the same volume normal saline for gavage.Examined the protein expression of aggrecanase4（ADAMTs-4） and aggrecanase5（ADAMTs-5） in cartilage tissue of knee.Examined the levels of interleukin 1β（IL-1β）,tumor necrosis factorα（TNF-α）,prostagladin E_2（PGE_2）,cyclooxygenase2（COX-2）,interferonγ（IFN-γ） in synovial membrane and serum.Results Compared with the normal group,the levels of IL-1β,TNF-α,PGE_2,COX-2,IFN-γin serum and synovial membrane of rats in the model group remarkably increased（P〈0.01）.Compared with the model group,the levels of IL-1β,TNF-α,PGE_2,COX-2,IFN-γ in serum and synovial membrane of rats in the Gulinggao group remarkably decreased（P〈0.01）.Compared with the Gulinggao group,the levels of IL-1β,TNFa,COX-2,IFN-γ in serum and the levels of IL-1β,COX-2,PGE_2,TNF-α,IFN-γ in synovial membrane of the Gugao group increased（P〈0.05 or P〈0.01）,the levels of IL-1β,TNF-α,COX-2,PGE_2,IFN-γ in serum and synovial membrane of the Linggao group increased（P〈0.05 or P〈0.01）.Also,the levels of IL-1β,TNF-α,PGE_2,IFN-γ in serum and IL-1β,PGE_2 in synovial membrane of the Celecoxib group increased（P〈0.01）.ADAMTs-4、ADAMTs-5 protein in knee cartilage tissue of rats in each group all expressed,of which the model group expressed the mostly remarkably.The Gulinggo group expressed the least.Conclusion Gulinggao may reduce osteoarthritis cartilage injury by the following mechanism：decreasing inflammatory cyctokines in serum and synovial membrane,and then preventing the synthesis of aggrecanase in local joint cartilage.And the effect was remarkably superior to its disassembled prescriptions and celecoxib.