温阳益髓方对膝骨关节炎模型大鼠膝关节软骨、骨代谢及NOX2/ROS/NF-κB水平的影响  

Effects of Wenyang Yisui decoction on cartilage,bone metabolism and NOX2/ROS/NF-κB activity in knee osteoarthritis rats

作　　者：李占军[1] 张志国[1] 宋巍 Li Zhanjun;Zhang Zhiguo;Song Wei(Department of Orthopedics,Daxing District People's Hospital,Beijing 102600,China)

机构地区：[1]北京市大兴区人民医院骨科,北京102600

出　　处：《解剖学杂志》2023年第4期285-289,共5页Chinese Journal of Anatomy

摘　　要：目的:探讨温阳益髓方对膝骨关节炎模型大鼠膝关节软骨、骨代谢及NOX2/ROS/NF-κB水平的影响。方法:选取40只SPF级SD雄性大鼠,随机分为正常组、模型组、低剂量温阳益髓方(低剂量)组、高剂量温阳益髓方(高剂量)组。除正常组外,其余3组均采用膝关节腔注射0.05 mL(20 mg/mL)碘乙酸钠,建立膝骨关节炎模型。建模成功后,低、高剂量组分别以6、24 mg/kg温阳益髓方汤剂灌胃,正常组、模型组以同体积生理盐水灌胃。采用H-E染色、光镜观察各组膝关节软骨组织的形态结构;ELISA法检测血清骨代谢相关指标水平;二氢乙锭荧光探针检测膝关节软骨组织活性氧(ROS)水平;免疫印迹检测膝关节软骨组织NADPH氧化酶2(NOX2)、核因子-κB(NF-κB)水平。结果:模型组膝关节软骨组织边缘的软骨基质减少、软骨囊结构不清、软骨细胞减少,且出现钙化灶。与模型组比较,低、高剂量组上述结构变化明显改善。与正常组比较,模型组Mankin评分显著升高;与模型组比较,低、高剂量组Mankin评分显著降低,且高剂量组低于低剂量组。与正常组比较,模型组血清骨钙素(OCN)、Ⅰ型胶原C末端肽(CTX-Ⅰ)水平显著降低,软骨寡聚基质蛋白(COMP)水平显著升高;与模型组比较,低、高剂量组血清OCN、CTX-Ⅰ水平显著升高,且高剂量组高于低剂量组,COMP水平显著降低,且高剂量组低于低剂量组。与正常组比较,模型组ROS水平显著升高;与模型组比较,低、高剂量组ROS水平显著降低,且高剂量组低于低剂量组。与正常组比较,模型组NOX2、NF-κB水平显著升高;与模型组比较,低、高剂量组NOX2、NF-κB水平显著降低,且高剂量组低于低剂量组。结论:温阳益髓方可改善膝骨关节炎模型大鼠膝关节软骨、骨代谢,其机制可能与其降低膝关节软骨组织NOX2/ROS/NF-κB水平有关。Objective:To investigate the effects of Wenyang Yisui decoction on knee cartilage,bone metabolism and NOX2/ROS/NF-κB level in knee osteoarthritis model rats.Methods:Forty SPF SD male rats were randomly divided into normal group,model group,low-dose Wenyang Yisui decoction(low-dose)group,and high-dose Wenyang Yisui decoction(high-dose)group.Except for the normal group,0.05 mL 20 mg/mL sodium iodoacetate were injected into the knee cavity to establish a knee osteoarthritis modelin rats of the remaining three groups.After successful modeling,the low-dose and high-dose groups were given 6 mg/kg and 24 mg/kg Wenyang Yisui decoction,respectively,while the normal group and model group were given the same volume of normal saline.The histopathologic morphology and structure of knee cartilage in each group were observed by H-E staining with light microscopy.Serum bone metabolism related indexes were detected by ELISA.The level of reactive oxygen species(ROS)in knee cartilage was detected by dihydroethidium fluorescent probe.The levels of NADPH oxidase 2(NOX2)and nuclear factor-κB(NF-κB)in knee cartilage were determined by Western blotting.Results:The model group showed reduced cartilage matrix,unclear chondrocapsule structure,reduced chondrocytes,and calcification foci at the edge of the knee cartilage tissue.Compared with the model group,the above structural changes were significantly improved in the low-dose group and the high-dose group.Compared with the normal group,Mankin score in model group were significantly increased,while Mankin score in low-dose and high-dose groups were significantly decreased compared with the model group,and high-dose group was lower than low-dose group.Compared with the normal group,the serum levels of osteocalcin(OCN)and C-terminal peptide of type Ⅰ collagen(CTX-Ⅰ)in model group were significantly decreased,while the levels of cartilage oligomerized matrix protein(COMP)were significantly increased.Compared with the model group,the serum levels of OCN and CTX-Ⅰ in low-dose and high-

关 键 词：温阳益髓方 膝骨关节炎 软骨 骨代谢 NADPH氧化酶2 活性氧 核因子-ΚB 大鼠 

分 类 号：R285.5[医药卫生—中药学]