乌头注射液对膝关节骨性关节炎模型兔体内COMP、p53蛋白、BMP-2等因子的影响  

Effects of Aconitum Injection on COMP,p53 Protein and BMP-2 in Knee Osteoarthritis Model Rabbits

作　　者：柳晓峰 安玉芳 崔立建 LIU Xiaofeng;AN Yufang;CUI Lijian(Heilongjiang Academy of TCM,Harbin 150036,China)

机构地区：[1]黑龙江省中医药科学院,哈尔滨150036

出　　处：《中国药房》2020年第6期714-718,共5页China Pharmacy

基　　金：哈尔滨市应用技术研究与开发项目(No.2017RAX-QJ056)。

摘　　要：目的:考察乌头注射液对膝关节骨性关节炎(KOA)模型兔体内软骨寡聚基质蛋白(COMP)、抑癌基因p53编码蛋白(p53蛋白)、骨形成蛋白2(BMP-2)等因子的影响,探讨该注射液治疗KOA的作用机制。方法:将24只兔随机分为空白组、模型组、玻璃酸钠组、乌头注射液组,每组6只。除空白组兔不作任何处理外,分别在第1、4、7天时于模型组、玻璃酸钠组、乌头注射液组兔关节腔注射2%木瓜蛋白酶溶液-0.03 mol/L L-半胱氨酸溶液以建立KOA模型;在造模成功后第1、4、7天时,分别于其关节腔注射生理盐水、玻璃酸钠注射液、乌头注射液各0.1 mL/kg。上述4组兔分别在给药结束后抽取关节液,采用酶联免疫吸附法测定COMP和p53蛋白含量;肉眼观察兔膝关节软骨形态;取膝关节软骨组织,采用苏木精-伊红法染色后于光镜下观察组织形态学改变并进行Mankin评分;采用PV二步法制作膝关节软骨组织的免疫组化标本,检测BMP-2的相对表达量。结果:与空白组比较,模型组兔膝关节软骨边缘破坏,软骨面有缺损;组织形态学观察结果显示,膝关节软骨组织结构明显不规则,软骨细胞可见明显扩大,有白泡样改变,细胞间分布紊乱,且Mankin评分显著升高(P<0.05);膝关节液中COMP、p53蛋白含量显著升高,膝关节软骨组织中BMP-2相对表达量显著降低(P<0.05)。与模型组比较,各给药组兔膝关节软骨的外观和组织形态学变化均有所改善,Mankin评分显著降低(P<0.05);膝关节液中COMP、p53蛋白含量显著降低,膝关节组织中BMP-2相对表达量显著升高(P<0.05),但乌头注射液组与玻璃酸钠组比较差异无统计学意义(P>0.05)。结论:乌头注射液具有改善KOA模型兔膝关节滑膜炎症、延缓关节软骨退变、保护关节的作用;其机制可能与抑制COMP分泌、降低p53蛋白表达、促进BMP-2释放有关。OBJECTIVE:To investigate the effects of Aconitum injection on cartilage oligomeric matrix protein(COMP),encoded protein by tumor suppressor gene p53(p53 protein) and bone morphogenetic protein 2(BMP-2) in knee osteoarthritis(KOA)model rabbit,so as to explore the mechanism of the injection in the treatment of KOA. METHODS:Totally 24 rabbits were randomly divided into blank group,model group,Sodium hyaluronate group and Aconitum injection group,with 6 rabbits in each group. KOA model was established by injecting 2% papain-0.03 mol/L L-cysteine solution into the articular cavity of rabbits in model group,sodium hyaluronate group and Aconitum injection group at the 1 st,4 th and 7 th day,respectively,except the blank group. At the 1 st,4 th and 7 th day after modeling succeeded,0.1 mL/kg of normal saline,Sodium hyaluronate injection and Aconitum injection were injected into the articular cavity of rabbits,respectively. The cartilage tissue of knee joint was taken from above 4 groups,and the contents of COMP and p53 protein were detected by ELISA. The cartilage morphology of rabbit knee joint was observed by naked eye. The cartilage of the knee joint was collected and stained by HE staining,and then the histomorphology changes were observed by light microscope;Mankin scoring was conducted. The two-step method of PV was used to make the immunohistochemical specimens of knee joint cartilage,and the relative expression of BMP-2 was detected. RESULTS:Compared with blank group,the edge of cartilage was damaged and the cartilage surface was damaged in the model group. The results of histomorphology observation showed that the joint tissue structure was obviously irregular,the distribution of chondrocytes was disordered with morphological changes,and the Mankin score was significantly increased(P<0.05);the contents of COMP and p53 protein in knee joint fluid were increased significantly,while the relative expression of BMP-2 in knee joint tissue decreased significantly(P<0.05). Compared with model group, the appearance, hist

关 键 词：乌头注射液 膝关节骨性关节炎 软骨寡聚基质蛋白 P53蛋白 骨形态发生蛋白2 兔 

分 类 号：R274.9[医药卫生—中西医结合] R965[医药卫生—中医骨伤科学]