摘要
目的探讨寒湿痹阻证膝骨性关节炎(knee osteoarthritis,KOA)大鼠模型制备方法。方法选取24只雄性SD大鼠,随机分为假手术组、手术组、寒湿痹阻证组,每组8只。假手术组右膝皮肤切开后立即缝合;手术组行前交叉韧带切断术(Anterior cruciate ligment transection,ACLT)切断前交叉韧带;寒湿痹阻证组在ACLT的基础上,术后第14天开始使用人工气候箱,温度为10.5℃,湿度90%,每天坚持同一时间段放置4 h,连续4周。4周后取材,通过Micro-CT观察大鼠膝关节骨骼微结构改变,酶联免疫吸附试验(Enzyme linked immunosorbent assay,ELISA)检测各组大鼠血液中白细胞介素-1β(Interleukin-1β,IL-1β)、白细胞介素-6(Interleukin-6,IL-6)、肿瘤坏死因子-α(Tumor necrosis factor-α,TNF-α)含量。结果与假手术组及手术组比较,寒湿痹阻证组出现毛发枯槁,出现畏寒、精神不振,反应敏感、畏惧等证型表现。与假手术组比较,手术组与寒湿痹阻证组大鼠膝周直径均增大,差异均有统计学意义(P<0.05)。术后1周,与假手术组比较,寒湿痹阻证组机械刺激缩足反应阈值(Paw with drawl threshold,PWT)和热刺激缩足潜伏期(Paw with drawal latency,PWL)均明显降低,差异有统计学意义(P<0.05)。术后3周、4周,与假手术组比较,手术组和寒湿痹阻证组均明显降低,差异有统计学意义(P<0.05)。Micro-CT显示,与假手术组比较,手术组、寒湿痹阻证组软骨表面粗糙不平、关节间隙狭窄,关节表层结构破坏。与假手术组比较,手术组、寒湿痹阻证组IL-1β、IL-6、TNF-α含量均显著增高,差异均有统计学意义(P<0.05)。结论本研究采用ACLT结合气候箱模拟寒湿痹阻证造模方法,兼顾中西医病因,模型评价全面,可为寒湿痹阻证KOA疾病的中医药现代化研究提供稳定模型。
Objective To explore the preparation method of a rat model of knee osteoarthritis(KOA)with cold dampness obstruction syndrome.Methods 24 male SD rats were selected as research subjects and randomly divided into sham surgery group,surgery group and cold dampness obstruction syndrome group,with 8 rats in each group.The skin of the right knee in the sham surgery group was cut open and immedi-ately sutured;The surgical group underwent anterior cruciate ligament transfer(ACLT)to cut the anteri-or cruciate ligament.On the basis of ACLT,the group with cold dampness obstruction syndrome started using an artificial climate chamber 14 days after surgery,with a temperature of 10.5℃and a humidity of 90%.The chamber was placed for 4 hours at the same time every day for 4 consecutive weeks.Observe the general condition of rats before modeling and at 1,3 and 4 weeks after modeling 4 weeks later,sam-ples were taken to observe the microstructural changes in the knee joint bones of rats using Micro CT.En-zyme linked immunosorbent assay(ELISA)was used to detect the levels of interleukin-1β(IL-1β),inter-leukin-6(IL-6)and tumor necrosis factor-α(TNF-α)in the blood of each group of the rats.Results It can be observed that compared with the sham surgery group and the surgery group,the cold dampness ob-struction syndrome group showed traditional Chinese medicine(TCM)syndrome manifestations of cold dampness obstruction.Compared with the sham surgery group,the knee circumference of the rats in the surgery group and the cold dampness obstruction syndrome group was increased significantly,and the differences were statistically significant(P<0.05).1 week after surgery,compared with the sham sur-gery group,the paw with draw threshold(PWT)and Paw Withdraw Latency(PWL)of mechanical stim-ulation foot contraction response in the cold dampness obstruction syndrome group were significantly re-duced,and the difference was statistically significant(P<0.05).At 3 and 4 weeks after surgery,com-pared with the sham surgery group,both the surgery group and the cold dampness obstruction syndrome group showed significant reductions,with statistical significance(P<0.05).Micro CT showed that com-pared with the sham surgery group,the surgical group and the cold dampness obstruction syndrome group had rough and uneven cartilage surfaces,narrow joint spaces and damaged joint surface structures.Com-pared with the sham surgery group,the levels of IL-1β,IL-6 and TNF-αwere significantly increased in the surgery group and the cold dampness obstruction syndrome group,and the differences were statistical-ly significant(P<0.05).Conclusion The ACLT+climate chamber simulation method used in this study can take into account the traditional Chinese and Western medicine causes of disease formation,and the model evaluation is comprehensive.It can provide a stable disease model for the modernization of TCM re-search on KOA disease caused by cold dampness obstruction syndrome.
作者
摆雪
田育魁
郭蕾
杨子
王程
李静先
朱清广
刘俊昌
BAI Xue;TIAN Yukui;GUO Lei;YANG Zi;WANG Cheng;LI Jingxian;ZHU Qingguang;LIU Junchang(College of Traditional Chinese Medicine,Xinjiang Medical University,Urumqi 830017,China;Tuina Department,Fourth Clinical Medical College,Xinjiang Medical University,Urumqi 830000,China;Tuina Department,Yueyang Hospital of Integrated Traditional Chinese and Western Medicine Affiliated to Shanghai University of Chinese Medicine,Shanghai 200437,China;College of Health Sciences,University of Ottawa,Ottawa K1N 6N5,Canada)
出处
《新疆医科大学学报》
2025年第7期897-903,共7页
Journal of Xinjiang Medical University
基金
国家自然科学基金项目(82360979)
新疆维吾尔自治区科技创新团队(天山创新团队)项目(2022TSYCTD0008)。
关键词
膝骨性关节炎
寒湿痹阻证
大鼠模型
knee osteoarthritis(KOA)
cold dampness obstruction syndrome
rat model
作者简介
摆雪(1989-),女,在读博士,讲师,主治医师,研究方向:中西医结合治疗骨关节疾病;通信作者:刘俊昌,男,教授,主任医师,博士生导师,研究方向:中西医结合防治骨关节疾病的基础与临床研究。
