基于以方测证的膝骨关节炎风寒湿痹证模型大鼠构建研究

来源:用户上传      作者:余晓愉　熊辉　齐新宇　向黎黎　曾朋　张娟

　　 〔摘要〕 目的 基于“以方y证”思维，模拟人膝骨关节炎（knee osteoarthritis， KOA）风寒湿痹证的发病机制，以期建立稳定的KOA病证结合动物模型。方法 将50只大鼠随机分为空白（A）组、单纯风寒湿刺激（B）组、单纯KOA造模（C）组、KOA造模+风寒湿刺激（D）组、乌头汤灌胃（E）组，共5组，每组10只。采用膝关节腔注射木瓜蛋白酶复合风寒湿外环境造模，基于“以方测证”思维运用乌头汤进行验证，灌胃结束后观察大鼠一般情况、膝关节软骨HE染色及Mankin评分、膝关节Micro-CT检测、血清肿瘤坏死因子-α（tumour necrosis factor-α， TNF-α）、白细胞介素-1β（interleukin-1β， IL-1β）、白细胞介素-17A（interleukin-17A， IL-17A）浓度。结果 Mankin评分显示，A、B组得分为0～1分，属于正常软骨;C、E组为4～7分，属KOA早中期;D组为7～11分，属于KOA中晚期。C、D、E组Mankin评分及TNF-α、IL-1β、IL-17A浓度较A组及B组均明显升高（P<0.05），C、E组Mankin评分及TNF-α、IL-1β、IL-17A浓度较D组均明显降低（P<0.05），B组TNF-α、IL-1β、IL-17A浓度较A组明显升高（P<0.05）。结论 与单纯KOA模型比较，采用膝关节腔注射木瓜蛋白酶复合风寒湿外环境刺激，KOA病情的进展更快，程度更严重，模型大鼠出现类似人体KOA风寒湿痹证的表现，并且乌头汤能够减轻复合模型风寒湿干预因素的影响，缓解疾病的进程。
　　 〔关键词〕 膝骨关节炎;风寒湿痹证;乌头汤;病证结合模型;Mankin评分;肿瘤坏死因子-α;白细胞介素-1β;白细胞介素-17A
　　 〔中图分类号〕R255.6 〔文献标志码〕A 〔文章编号〕doi：10.3969/j.issn.1674-070X.2022.07.007
　　Study on establishment of knee osteoarthritis model rats with wind-cold-dampness
　　syndrome based on testing syndrome by formula
　　YU Xiaoyu XIONG Hui QI Xinyu XIANG Lili ZENG Peng ZHANG Juan
　　（1. Hunan Chest Hospital， Changsha， Hunan 410005， China; 2. Hunan University of Chinese Medicine， Changsha， Hunan 410208， China; 3. The Second Affiliated Hospital of Hunan University of Chinese Medicine， Changsha， Hunan 410005， China;
　　4. Xiangxing College of Hunan University of Chinese Medicine， Xiangyin， Hunan 414600， China）
　　〔Abstract〕 Objective To simulate the pathogenesis of human knee osteoarthritis （KOA） with wind-cold-dampness syndrome based on the thinking of "testing syndrome by formula"， so as to establish a stable animal model of KOA combination of disease and syndrome. Methods Fifty rats were randomly divided into blank （A） group， simple wind-cold-dampness stimulation （B） group， simple KOA model （C） group， KOA model+wind-cold-dampness stimulation （D） group and Wutou Decoction intragastric administration （E） group， with 10 rats in each group. The model was made by injecting papain into the knee cavity combined with the external environment of wind， cold and dampness， and verified by Wutou Decoction based on the thinking of testing syndrome by formula. After the end of ingastric administration， the general situation of the rats， HE staining of knee cartilage， the Mankin score， the knee micro-CT detection and the tumor necrosis factor-α （TNF-α）， interleukin-1β （IL-1β）， interleukin-17A （IL-17A） concentration were observed. Results Mankin score showed that the scores of group A and B were 0-1， belonging to normal cartilage; the scores of group C and E were 4-7， belonging to early and middle KOA; the score of group D was 7-11， belonging to the middle and late KOA. Mankin score and the content of TNF-α， IL-1β and IL-17A of group C， D and E were higher than those of group A and B （P<0.05）， Mankin score and the content of TNF-α， IL-1β and IL-17A of group C and E were lower than those of group D （P<0.05）. The content levels of TNF-α， IL-1β and IL-17A of group B were higher than those of group A （P<0.05）. Conclusion Compared with the simple KOA model， the progress of KOA disease is faster and more serious by injecting papain into the knee cavity combined with the external environment of wind， cold and dampness. The model rats appear similar to the performance of human body KOA with wind-cold-dampness syndrome， and Wutou Decoction can reduce the influence of wind， cold and dampness intervention factors in the composite model and alleviate the process of disease.

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