加味独活寄生合剂对兔膝骨关节炎模型软骨TLR4/MyD88/NF-kB信号通路的影响  

Effect of Modified Duhuo Jisheng Mixture on TLR4/MyD88/NF-κB Signaling Pathway in Cartilage of the Rabbit Model of Knee Osteoarthritis

作　　者：曹寅生[1] 邝高艳[1] 危建文 金久楚 刘雄 朱付平[1] CAO YIN-sheng;KUANG Gao-yan;WEI Jian-wen;JIN Jiu-chu;LIU Xiong;ZHU Fu-ping(The First Affiliated Hospital of Hunan University of Chinese Medicine,Changsha Hunan 410007;Hunan University of Chinese Medicine,Changsha Hunan 410208)

机构地区：[1]湖南中医药大学第一附属医院,湖南长沙410007 [2]湖南中医药大学,湖南长沙410208

出　　处：《世界中西医结合杂志》2023年第7期1273-1277,共5页World Journal of Integrated Traditional and Western Medicine

基　　金：湖南省自然科学基金项目(2020JJ5422);湖南省中医药科研计划项目(2021088)。

摘　　要：目的观察加味独活寄生合剂对兔膝骨关节炎软骨组织中TLR4,MyD88,NF-κB表达的影响,探讨加味独活寄生合剂减缓关节软骨退变的可能作用机理。方法将35只6月龄雄性新西兰兔按随机数字表法分为25只KOA造模组和10只正常组。用改良Videman造模法制备兔KOA模型。造模成功后,从正常组中随机选取6只实验兔作为空白组,从KOA造模组中随机选取18只KOA造模实验兔按随机分配的方式分为模型组、硫酸氨基葡萄糖组以及加味独活寄生合剂组,各组各6只。空白组、模型组予蒸馏水灌胃,硫酸氨基葡萄糖组予硫酸氨基葡萄糖混悬液灌胃,加味独活寄生合剂组予加味独活寄生合剂灌胃,疗程均为6周。给药结束,HE染色观察关节软骨并进行Mankin评分评估;WesternBlot法检测其TLR4、MyD88、NF-κB蛋白的表达;RT-PCR法检测其TLR4、MyD88、NF-κBmRNA的表达。结果模型组与空白组比较,膝关节软骨破坏更为明显,Mankin评分升高,差异有统计学意义(P<0.01),软骨中MyD88、TLR4、NF-κBmRNA及蛋白表达水平均有所上调,差异有统计学意义(P<0.01)。加味独活寄生合剂组及硫酸氨基葡萄糖组与模型组比较,软骨损伤程度较轻,Mankin评分降低,差异有统计学意义(P<0.01),软骨中TLR4、MyD88、NF-κBmRNA及蛋白表达水平均有所下调,差异有统计学意义(P<0.01)。结论兔KOA关节软骨退变能被加味独活寄生合剂有效延缓,其作用机制可能与抑制软骨组织TLR4/MyD88/NF-κB信号通路的活化,从而减轻炎症反应相关。Objective To observe the effects of modified Duhuo Jisheng Mixture on the expression of Toll-like receptor 4(TLR4),myeloid differentiation factor 88(MyD88),nuclear factor-kappa B(NF-κB)in the cartilage of rabbit model of knee osteoarthritis(KOA),so as to explore the possible mechanism of modified Duhuo Jisheng Mixture in slowing down the degeneration of articular cartilage.Methods Thirty-five six-month-old male New Zealand rabbits were randomized into a normal group(n=10)and a KOA modeling group(n=25)according to the random number table method.After successful modeling by the modified Videman method,6rabbits were randomly selected from the normal group as the blank group,and 18 rabbits were randomly selected from the KOA modeling group and randomized into model,glucosamine sulfate,and modified Duhuo Jisheng Mixture groups(n=6).The rabbits in the blank group and model group were administrated with distilled water by gavage,and those in other groups with corresponding agents by gavage.The course of treatment was 6 weeks.After administration,the articular cartilage was stained with hematoxylin-eosin(HE)and evaluated based on the Mankin score.Western blotting and RT-PCR were employed to determine the protein and mRNA levels,respectively,of TLR4,MyD88,and NF-κB.Results Compared with the blank group,the modeling damaged the cartilage of knee joint,increased the Mankin score(P<0.01),and up-regulated the mRNA and protein levels of TLR4,MyD88,NF-κB in the cartilage(P<0.01).Compared with the model group,modified Duhuo Jisheng Mixture and glucosamine sulfate alleviated the cartilage damage,lowered the Mankin score(P<0.01),and down-regulated the mRNA and protein levels of TLR4,MyD88,and NF-κB in the cartilage(P<0.01).Conclusion Modified Duhuo Jisheng Mixture can effectively delay the degeneration of articular cartilage in the rabbit model of KOA by inhibiting the activation of the TLR4/MyD88/NF-κB signaling pathway to reduce inflammation.

关 键 词：膝骨关节炎 加味独活寄生合剂 TLR4 MYD88 NF-κB 

分 类 号：R285.5[医药卫生—中药学]