摘要
目的观察加味独活寄生合剂对膝骨关节炎(KOA)模型兔软骨OPG/RANKL轴及Ⅱ型胶原表达的影响,丰富加味独活寄生合剂治疗膝骨关节炎的作用机制。方法将3月龄新西兰雄兔24只,随机分为空白组、模型组、加味独活组和塞来昔布组,每组6只。采用Ⅱ型胶原酶膝关节腔内注射法建立兔KOA模型。造模2周后开始给药,给药时间共计6周。采用番红固绿染色法观察各组软骨形态结构改变,免疫荧光法检测各组软骨中OPG、RANKL、Ⅱ型胶原蛋白表达水平,qPCR法检测OPG、RANKL、Ⅱ型胶原mRNA水平。结果①番红固绿染色结果:与模型组相比,加味独活组Mankin′s评分显著降低(P<0.01)。②免疫荧光结果:加味独活组OPG/RANKL的平均光密度值最高,与塞来昔布组及模型组相比,差异有显著统计学意义(P<0.01)。③qPCR结果:qPCR检测显示加味独活组OPG mRNA水平较其他各组显著上调(P<0.01),OPG/RANKL mRNA水平较其他各组显著上调(P<0.01)。结论加味独活寄生合剂可有效促进膝骨关节炎兔软骨修复,其作用机制可能和调节OPG/RANKL轴并促进Ⅱ型胶原修复有关。
Objective To observe the effect of Jiawei Duhuo Jisheng Mixture(加味独活寄生合剂)on the ostoeprotegerin(OPG)/receptor activator of NF-κB ligand(RANKL)and collagen Ⅱ expression in cartilage of rabbits with knee osteoarthritis(KOA)and to enrich the mechanism of action of Jiawei Duhuo Jisheng Mixture in treating KOA.Methods Twenty-four 3-month-old New Zealand male rabbits were randomly divided into a blank group,a model group,a Jiawei Duhuo Mixture group and a celecoxib group,with 6 rabbits in each group.A rabbit KOA model was established by injecting collagenase Ⅱ into the knee joint cavity.The administration started after 2 weeks of modeling.After administration,the morphological and structural changes of cartilage in each group were observed by using safranin green staining method,the expression levels of OPG,RANKL and type Ⅱ collagen in each group′s cartilage were detected by using immunofluorescence method,and the mRNA levels of OPG,RANKL and type Ⅱ collagen were detected by using qPCR method.Results①SafraninO-staining result:Compared with that of the model group,the Mankin′s score of the Jiawei Duhuo Mixture group was significantly reduced(P<0.01).②Immunofluorescence results:The average optical density of OPG/RANKL in the Jiawei Duhuo Mixture group was the highest,and the difference was statistically significant(P<0.01)compared to that in the celecoxib group and model group.③The qPCR results:The qPCR detection showed that the OPG mRNA level in the Jiawei Duhuo Mixturegroup was significantly up-regulated compared to that in the other groups(P<0.01),and the OPG/RANKL mRNA level was significantly up-regulated compared to that in the other groups(P<0.01).Conclusion Jiawei Duhuo Jisheng Mixture can effectively promote cartilage repairing in KOA rabbits and delay the progression of KOA.Its mechanism of action may be related to up-regulating type Ⅱ collagen and OPG/RANKL axis.
作者
陈树源
龚亮宇
许晓彤
卢敏
CHEN Shuyuan;GONG Liangyu;XU Xiaotong;LU Min(Xiangtan Traditional Chinese Medicine Hospital,Xiangtan 411100,Hunan,China;First Hospital of Hunan University of Chinese Medicine,Changsha 410007,Hunan,China)
出处
《辽宁中医杂志》
北大核心
2025年第4期183-188,I0006,I0007,共8页
Liaoning Journal of Traditional Chinese Medicine
基金
国家自然科学基金面上项目(81874476)
国家中医药管理局全国名老中医药专家传承工作室建设项目(国中医药人教函〔2022〕75号)
湖南省中西医结合损伤临床医疗技术示范基地项目(2023SK4064)。
作者简介
陈树源(1997-),男,湖南湘潭人,硕士,研究方向:骨伤疾病的中医药防治;通讯作者:卢敏(1962-),男,湖南长沙人,教授、主任医师,博士研究生导师,研究方向:骨与关节损伤及修复,E-mail:lumin6563@163.com。
