蠲痹胶囊不同提取部位对绝经后膝骨关节炎大鼠的影响

Effects of different extraction parts of Juanbi Capsules on postmenopausal rats with knee osteoarthritis

目的:观察蠲痹胶囊各提取部位对膝骨关节炎(knee osteoarthritis,KOA)模型大鼠的作用,筛选其药效主要活性部位。方法:使用系统溶剂提取法获得蠲痹胶囊乙醇、水、石油醚、三氯甲烷、正丁醇和乙酸乙酯提取活性部位,并以改良Hulth法并摘除双侧卵巢建立大鼠绝经后KOA模型为研究对象。实验大鼠被随机分为不同组别:正常对照组、模型对照组以及蠲痹胶囊乙醇提取部位组、水提取部位组、石油醚提取部位组、三氯甲烷提取部位组、正丁醇提取部位组、乙酸乙酯各提取部位组,观察这些组在各提取部位的干预下,膝骨关节炎大鼠膝关节横向直径的变化,使用micro-CT分析膝关节骨微结构;利用ELISA法检测血清、关节液中的相关炎症介质TNF-α、IL-1β、MMP9水平;通过Western blot检测软骨组织中MMP9、COX-2蛋白表达水平。结果:与模型对照组相比,蠲痹胶囊乙醇提取部位较其他部位更加显著降低KOA大鼠膝关节肿胀;有效改善大鼠膝关节骨小梁相对体积、骨小梁厚度、骨小梁数量等骨微结构变化;能明显降低大鼠血清、关节液中TNF-α、IL-1β、MMP9水平,抑制软骨中MMP9、COX-2蛋白的表达水平。结论:蠲痹胶囊各提取部位均可不同程度缓解KOA大鼠关节炎症状,其中乙醇提取部位是其药效主要活性部位。

OBJECTIVE To evaluate the effects of Juanbi Capsules(JC)in the prevention and treatment of knee osteoarthri⁃tis(KOA)and screen the major active parts of its efficacy.METHODS The extraction parts of ethanol,water,petroleum ether,trichloromethane,n-butanol and ethyl acetate of JC were acquired through systematic solvent extraction.Postmenopausal rat KOA model was established by modified Hulth method after bilateral ovariectomy.The rats were randomized into three groups of control,model control and Juanbi capsule ethanol extract group,water extract group,petroleum ether extract group,trichloro⁃methane extract group,n-butanol extract group,ethyl acetate extract group.The changes of transverse diameter of knee joint of KOA rats were observed under the intervention of different extraction sites.Micro-CT was performed for examining bone micro⁃structure of knee joint.The levels of tumor necrosis factor alpha(TNF-α),interleukin-1β(IL-1β)and matrix metalloproteinase 9(MMP9)in sera and joint fluid were detected by enzyme-linked immunosorbent assay(ELISA).The expression levels of MMP9 and cyclooxygenase-2(COX-2)in cartilage tissue were detected by Western blot.RESULTS As compared with model control group,JC ethanol extract site significantly lessened knee swelling in KOA rats.Relative volume,thickness and number of bone trabeculae of knee joint effectively improved.It could significantly lower the levels of TNF-α,IL-1βand MMP9 in serum and joint fluid.And the expression levels of MMP9 and COX-2 related inflammatory mediators declined markedly in cartilage.CONCLUSION All extraction parts of JC may relieve arthritic symptoms of KOA rats to varying degrees.And ethanol extract part is a major active ingredient.