摘要
目的:观察六味骨痹汤对兔骨关节炎模型关节软骨形态、Ⅱ型胶原蛋白表达及体液中胶原酶3(MMP-13)、肿瘤坏死因子-α(TNF-α)、软骨寡聚基质蛋白(COMP)的影响,探讨其对骨关节炎(OA)软骨的修复作用及其可能机制。方法:取2~3月龄清洁级新西兰兔32只,体质量(2.42±0.23)kg,采用木瓜蛋白酶关节注射法建立兔OA模型,随机区组法分为六味骨痹汤组、硫酸氨基葡萄糖(GS)组和模型组,另设正常组。前两组分别予六味骨痹汤3.83g/(kg·d)、硫酸氨基葡萄糖胶囊77mg/(kg·d)溶于生理盐水10mL灌胃,模型组和正常组均予生理盐水(10mL/d)灌胃。6周后采用酶联免疫吸附法(ELISA)检测血清及关节液中MMP-13,TNF-α及COMP的含量;处死实验兔,按Pelletier原则对关节软骨大体形态评分;取软骨病理标本,部分予苏木精-伊红染色后用改良Mankin's法予组织形态学观察评分,其他标本免疫组化染色后对其Ⅱ型胶原蛋白表达情况进行观察评分。结果:与模型组相比,六味骨痹汤组血清和关节液中MMP-13,TNF-α及COMP的含量降低,差异有统计学意义(P<0.05);关节软骨Pelletier评分和改良Mankin's评分显著降低,差异有统计学意义(P<0.05);Ⅱ型胶原蛋白表达有所增加,差异有统计学意义(P<0.05).与GS组相比,上述指标差异均无统计学意义(P>0.05).结论:六味骨痹汤可促进兔OA模型软骨修复,其机制可能是通过降低兔血清和关节液中MMP-13,TNF-α等因子的含量抑制软骨破坏,从而提高Ⅱ型胶原蛋白的表达来发挥作用。
Objective：To observe the morphology of articular cartilage,the expression of typeⅡcollagen and the collagenase 3（MMP-13）,tumor necrosis factor-α（TNF-α）and cartilage oligomeric matrix protein（COMP）in rabbit osteoarthritis model and explore the effect of Liuweigubi Decoction on osteoarthrosis（OA）cartilage and its possible mechanism in rabbits with osteoarthritis.Methods：Thirty-two clean-grade New Zealand rabbits aged 2to 3 months were used and the body weight was（2.42±0.23）kg.Rabbit OA models which established by papain joint injection were randomly divided into Liuweigubi decoction（LD）group,glucosamine sulfate（GS）group and model group.In addition,normal group was set up separately.The first two groups were respectively treated with Liuweigubi decoction 3.83g/（kg·d）and glucosamine sulfate capsules 77 mg/（kg·d）which were dissolved in 10 mL normal saline.Normal group and model group received normal saline 10（mL/d）gavage.After 6weeks,serum MMP-13,TNF-α,and COMP levels in serum and joint fluid were measured by enzyme-linked immunosorbent assay（ELISA）.Rabbits were sacrificed,and general articular cartilage morphology was scored according to the Pelletier principle.Cartilage pathological specimens were taken,and after hematoxylin-eosin staining,histomorphological observations were performed using improved Mankin＇s method.The expression of typeⅡcollagen was observed in other specimens after immunohistochemical staining.Results：Compared with the model group,the levels of MMP-13,TNF-αand COMP in the serum and synovial fluid of the Liuweigubi decoction group decreased（P〈0.05）,and the Pelletier score and improved Mankin＇s score of the articular cartilage decreased significantly（P〈0.05）.The expression of typeⅡcollagen was increased（P〈0.05）.There was no statistically significant difference between the above parameters compared with GS group（P〈0.05）.Conclusion：Liuweigubi decoction can promote the repair of OA model cartilage in rabbits.The mechanism may be to reduce cartilage destruction by decreasing the content of MMP-13,TNF-αand other factors in rabbit serum and synovial fluid,so as to increase the expression of type Ⅱ collagen.
作者
李媛
陈达
余阗
彭力平
胡烈奎
廖州伟
马笃军
LI Yuan;CHEN Da;YU Tian;PENG Liping;HULiekui;LIAO Zhouwei;MA Dujun(Fourth Clinical Medical College,Affiliated to Guangzhou University of Traditional Chinese Medicine,Shenzhen 518033,Guangdong China;Baoan Hospital of Traditional Chinese Medicine,Shenzhen 518101,Guangdong China;Shenzhen Hospital of Traditional Chinese Medicine,Shenzhen 518033,Guangdong China.)
出处
《中国中医骨伤科杂志》
CAS
2018年第9期9-13,18,共6页
Chinese Journal of Traditional Medical Traumatology & Orthopedics
基金
广东省中医药局科研项目(20171245)
关键词
六味骨痹汤
骨关节炎
兔
关节软骨修复
Ⅱ型胶原蛋白
Liuweigubi decoction
osteoarthritis
rabbit
articular cartilage repair
type Ⅱ collagen
