小鼠膝骨关节炎阳虚血瘀证模型的构建与评价  

Construction and evaluation of a mouse model of knee osteoarthritis with yang-deficiency and blood-stasis syndrome

作　　者：蔡宏杰 王旭 徐建波 林士能 童培建[1] 金红婷[1] 陈佳丽 CAI Hongjie;WANG Xu;XU Jianbo;LIN Shineng;TONG Peijian;JIN Hongting;CHEN Jiai(Zhejiang Provincial Hospital of Traditional Chinese Medicine,Hangzhou 310006,Zhejiang,China;The First Clinical Medical College of Zhejiang Chinese Medical University,Hangzhou 310053,Zhejiang,China)

机构地区：[1]浙江省中医院,浙江杭州310006 [2]浙江中医药大学第一临床医学院,浙江杭州310053

出　　处：《中医正骨》2023年第6期18-23,共6页The Journal of Traditional Chinese Orthopedics and Traumatology

基　　金：国家自然科学基金项目(81873324);浙江中医药大学基本科研能力提升项目(2021JKJNTZ014A)。

摘　　要：目的:探讨构建小鼠膝骨关节炎(knee osteoarthritis,KOA)阳虚血瘀证模型的有效方法。方法:将12只10周龄雄性无特定病原C57BL/6J小鼠采用内侧半月板失稳(destabilized medial meniscus,DMM)法构建KOA模型。KOA造模成功后,将12只KOA模型小鼠随机分为KOA阳虚血瘀证组和KOA对照组,每组6只。KOA阳虚血瘀证组小鼠于浸在冰水混合物中的塑料鼠箱中饲养,并以冰水混合物为饮用水;KOA对照组小鼠于22℃干燥环境中正常饲养。饲养10周后,评价小鼠阳虚血瘀证表现,分析小鼠步态,处死小鼠后进行小鼠膝关节MicroCT分析、膝关节组织病理学观察及膝关节软骨Ⅱ型胶原蛋白α1链表达检测。结果:①小鼠阳虚血瘀证评价结果。KOA阳虚血瘀证组小鼠的阳虚证评分和血瘀证评分均高于KOA对照组[(13.33±1.37)分,(3.50±1.23)分,t=13.120,P=0.000;(4.83±0.75)分,(0.83±0.75)分,t=9.200,P=0.000]。②小鼠步态分析结果。2组各收集到5只小鼠完整的步态分析数据。KOA阳虚血瘀证组小鼠右后肢的爪面积和步幅均小于KOA对照组[(0.610±0.094)cm2,(0.758±0.042)cm2,t=-3.211,P=0.012;(1.780±0.045)cm,(2.040±0.152)cm,t=-3.677,P=0.006];2组小鼠右后肢的摆动期时间和站立期时间比较,组间差异均无统计学意义[(0.060±0.004)s,(0.060±0.009)s,t=-0.096,P=0.963;(0.062±0.009)s,(0.076±0.014)s,t=-1.810,P=0.137]。③小鼠膝关节MicroCT分析结果。KOA阳虚血瘀证组小鼠的骨体积分数低于KOA对照组[(55.582±2.810)%,(69.248±1.884)%,t=-6.997,P=0.002],骨小梁分离度大于KOA对照组[(0.114±0.009)mm,(0.077±0.010)mm,t=4.883,P=0.008];2组小鼠的骨小梁厚度、骨小梁数量比较,组间差异均无统计学意义[(0.148±0.005)mm,(0.210±0.036)mm,t=-2.939,P=0.094;(3.767±0.100)个·mm^(-1),(3.368±0.547)个·mm^(-1),t=1.243,P=0.282]。④小鼠膝关节组织病理学观察结果。2组小鼠膝关节软骨均有缺失,且KOA阳虚血瘀证组小鼠膝关节软骨缺失较KOA对照组更严重。⑤小Objective:To explore the effective method for building a yangdeficiencybloodstasistype knee osteoarthritis(KOA)mouse model.Methods:Twelve 10weekold specific pathogen free(SPF)grade male C57BL/6J mice were selected out and were intervened by destabilized medial meniscus(DMM)method for inducing KOA.After successful modeling,the 12 KOA model mice were randomly divided into yangdeficiencybloodstasistype KOA group and KOA control group,6 cases in each group.The mice in yangdeficiencybloodstasistype KOA group were bred in plastic box submerged in icewater mixture,and they were fed with icewater mixture,while the ones in KOA control group were bred normally in a dry environment of 22℃.After 10week feeding,the yangdeficiency and bloodstasis syndrome in KOA model mice were observed and evaluated,and the gaits were analyzed.After that,the mice were executed by using cervical dislocation method,and the knee samples were harvested from their right hind limbs.The knee samples were scanned and analyzed by using microCT and CTAn1.10 software respectively,followed by observation on histopathological changes of knee tissues and determination of the expression levels of typeⅡcollagenα1 chain(Col2A1)in knee cartilage.Results:①The scores of yangdeficiency syndrome and bloodstasis syndrome were higher in yangdeficiencybloodstasistype KOA group compared to KOA control group(13.33±1.37 vs 3.50±1.23 points,t=13.120,P=0.000;4.83±0.75 vs 0.83±0.75 points,t=9.200,P=0.000).②The complete gait analysis data was collected from 5 mice in each group.The paw area and stride length of the right hind limbs were smaller in yangdeficiencybloodstasistype KOA group compared to KOA control group(0.610±0.094 vs 0.758±0.042 cm(2),t=-3.211,P=0.012;1.780±0.045 vs 2.040±0.152 cm,t=-3.677,P=0.006).There was no statistical difference in the time of swing phase and stance phase of the right hind limbs between the 2 groups(0.060±0.004 vs 0.060±0.009 seconds,t=-0.096,P=0.963;0.062±0.009 vs 0.076±0.014 seconds,t=-1.810,P=0.137).③The bone vol

关 键 词：骨关节炎 膝 小鼠 疾病模型 动物 阳虚血瘀 动物实验 

分 类 号：R274.9[医药卫生—中西医结合] R-332[医药卫生—中医骨伤科学]