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目的观察复元胶囊对实验性大鼠晚期膝骨关节炎软骨组织中基质金属蛋白酶1(MMP-1)、MMP-3及Ⅱ型胶原的影响,探讨其治疗骨关节炎的作用机制。方法成年雄性SD大鼠60只,随机分为六组,每组10只。除正常组外,其余各组均采用Hulth法建立晚期膝骨关节炎模型。手术后第2周开始,抗骨增生胶囊组予抗骨增生胶囊557 mg·kg-1·d-1,复元胶囊低、中、高剂量组分别给予复元胶囊371.65、743.30、2 229.90 mg·kg-1·d-1,正常组和模型组给予等体积生理盐水,每日灌胃1次,持续12周。12周后评价各组大鼠关节肿胀情况,并取大鼠胫骨平台的软骨组织。按Pelletier-JP标准评价关节软骨大体情况,并通过HE染色进行Mankin’s评分。采用免疫组织化学法和Western blot法检测各组关节软骨中MMP-1、MMP-3及Ⅱ型胶原的蛋白表达量。结果与正常组相比,模型组大鼠关节肿胀评分、PelletierJP标准和Mankin’s评分均升高(P<0.05或P<0.01),而各给药组关节肿胀评分、Pelletier-JP标准和Mankin’s评分均低于模型组(P<0.05或P<0.01)。复元胶囊中、高剂量组Mankin’s评分低于抗骨增生胶囊组(P<0.05或P<0.01)。与正常组相比,模型组软骨组织中MMP-1、MMP-3表达量明显升高,Ⅱ型胶原表达量明显降低(P<0.01)。各给药组MMP-1、MMP-3表达量低于模型组,Ⅱ型胶原表达量高于模型组(P<0.05或P<0.01)。复元胶囊高剂量组MMP-1、MMP-3的表达量低于抗骨增生胶囊组和复元胶囊低、中剂量组,Ⅱ型胶原表达量高于抗骨增生胶囊组和复元胶囊低、中剂量组(P<0.05或P<0.01)。结论复元胶囊能通过抑制MMPs的表达,减少胶原的降解,达到对晚期骨关节炎的防治作用。
Objective To observe the effect of Fuyuan Capsule on the expressions of matrix metalloproteinase-1 (MMP-1), MMP-3 and collagen Ⅱ in the cartilage tissue of advanced knee osteoarthritis in rats and to explore its mechanism of action in treating osteoarthritis. Methods Sixty adult male Sprague-Dawley rats were randomly divided into six groups of 10 rats. In addition to the normal group, the rest of the groups were established using Hulth method of advanced knee osteoarthritis model. At the second week after operation, the anti-bone-augmentation capsule group was given 557 mg · kg-1 · d-1 anti-bone-augmentation capsules and the low, middle and high dose groups were given Fuyuan capsules 371.65, 743.2 and 229.90 mg · kg- -1 · d-1, normal group and model group were given equal volume of saline, gavage once daily for 12 weeks. Twelve weeks later, the swollen joints of the rats in each group were evaluated, and the cartilage of the tibial plateau in rats was taken. The general condition of articular cartilage was evaluated according to the Pelletier-JP standard, and Mankin’s score was determined by HE staining. Immunohistochemistry and Western blot were used to detect the protein expression of MMP-1, MMP-3 and type Ⅱ collagen in articular cartilage of each group. Results Compared with the normal group, the joint swelling index, PelletierJP standard and Mankin’s score in the model group were significantly increased (P <0.05 or P <0.01), while the joint swelling score, Pelletier-JP standard and Mankin’s score Lower than model group (P <0.05 or P <0.01). Mankin’s score in Fuyuan capsule group was lower than that in Kangaojianjian Capsule group (P <0.05 or P <0.01). Compared with normal group, the expression of MMP-1 and MMP-3 in cartilage tissue of model group was significantly increased and the expression of type Ⅱ collagen was significantly decreased (P <0.01). The expression of MMP-1 and MMP-3 in each administration group was lower than that in model group, and the expression of type II collagen was higher than that in model group (P <0.05 or P <0.01). The expression of MMP-1 and MMP-3 in Fuyuan capsule high-dose group was lower than that in anti-bone-augmentation capsule group and Fuyuan capsule low- and middle-dose group, and the expression of typeⅡcollagen was higher than that in anti-bone-augmentation capsule group and Fuyuan capsule Group (P <0.05 or P <0.01). Conclusion Fuyuan capsule can inhibit the expression of MMPs and reduce the degradation of collagen to achieve the prevention and treatment of advanced osteoarthritis.