透骨消痛胶囊及其拆方对退变软骨细胞Wnt/β-catenin信号通路的影响  

Effect of Tougu Xiaotong Capsule and its disassembled recipes on chondrocyte degeneration of rats via Wnt/beta-catenin signaling pathway

作　　者：吴追乐[1] 陈星强[1] 陈春蓉[1] 于超[1] 朱晓勤[1] 庄志强[2] 

机构地区：[1]福建中医药大学,福建省福州市350122 [2]福建中医药大学附属漳州市中医院,福建省漳州市363000

出　　处：《中国组织工程研究》2016年第24期3568-3573,共6页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金资助项目(81202713);陈可冀中西医结合发展基金资助项目(CKJ2015010);福建省自然科学基金项目(2015J05161)~~

摘　　要：背景:前期研究表明,透骨消痛胶囊能调控软骨细胞Rho/Rock信号通路、抑制软骨细胞线粒体凋亡通路的信号转导、改变软骨下骨重塑的速率和模式、改善软骨下骨胶原纤维和钙磷结晶的排列等对骨性关节炎发挥治疗作用。目的:进一步观察透骨消痛胶囊及其拆方含药血清对大鼠退变软骨细胞Wnt/β-catenin信号通路的影响,揭示方中治疗骨性关节炎的主要治法。方法:(1)取SD大鼠40只随机分为透骨消痛胶囊组、补肾柔肝组、活血祛风组和生理盐水组,按照药理试验中动物间和动物与人体间的等效剂量换算,前3组分别灌服等效剂量的透骨消痛胶囊、补肾柔肝药、活血祛风药,生理盐水组灌服生理盐水,制备含药血清用于细胞实验。(2)体外培养SD大鼠软骨细胞并分为5组:空白对照组、模型组、透骨消痛胶囊组、补肾柔肝组和活血祛风组。模型组及各给药组用白细胞介素1β干预软骨细胞24 h后,加入各组血清干预软骨细胞72 h,空白对照组用盐水血清培养,同一时间收集空白对照组软骨细胞。(3)采用实时定量反转录-聚合酶链反应和Western blot法分析Wnt 4、β-catenin、基质金属蛋白酶13的表达。结果与结论:(1)与空白对照组相比,模型组Wnt 4、β-catenin、基质金属蛋白酶13表达均明显增加。(2)与模型组相比,透骨消痛胶囊组、补肾柔肝组和活血祛风组的Wnt 4、β-catenin、基质金属蛋白酶13表达均明显降低,且降低程度为透骨消痛胶囊组>补肾柔肝组>活血祛风组。(3)结果提示透骨消痛胶囊对白细胞介素1β诱导的软骨细胞具有协同保护作用,补肾柔肝为透骨消痛胶囊治疗骨性关节炎的主要治法。BACKGROUND：Previous studies have showed that Tougu Xiaotong Capsule（TGXTC） exerts better effects on osteoarthritis,by regulating Rho/Rock signaling pathway,inhibiting signal transduction of chondrocyte mitochondrial apoptosis pathway,varying the rate and pattern of subchondral bone remodeling and improving the arrangement of subchondral bone collagen fibers and calcium-phosphate crystallization.OBJECTIVE：To observe the effects of the serum containing TGXTC and its disassembled recipes on chondrocyte degeneration of rats via Wnt/β-catenin signal pathway,and to explore the main therapeutic method for osteoarthritis in the TGXTC.METHODS：Forty Sprague-Dawley rats were randomly assigned to receive the treatment of TGXTC,Bushen Rougan（BSRG）,Huoxue Qufeng（HXQF） and normal saline,respectively,according to the dose conversion methods of animal to animal and animal to human.Then various drug-containing serums were prepared for the following cellular experiment.After culture and passage,chondrocytes from Sprague-Dawley rats at passage 3 were divided into five groups：blank control,model,TGXTC,BSRG,HXQF groups.Cells in the latter four groups were cultured in appropriate drug-containing serums（normal saline serum for the model group） for 72 hours,following intervention with interleukin-1β for 24 hours.Cells in the blank control group were cultured in normal saline serum.Afterwards,cells in all the five groups were collected for detecting expression of Wnt 4,β-catenin and matrix metalloproteinase 13 at mR NA and protein levels using real-time PCR and western blot assay,respectively.RESULTS AND CONCLUSION：Compared with the blank control group,the expression of Wnt 4,β-catenin,matrix metalloproteinase 13 was significantly increased in the model group.Compared with the model group,the expression of Wnt 4,β-catenin,matrix metalloproteinase 13 in the TGXTC,BSRG and HXQF groups were decreased significantly,sequenced as TGXTC group BSRG group HXQF group.These results indicate that TGXTC plays a synergist

关 键 词：医学 中国传统 骨关节炎 软骨 组织工程 组织构建 软骨细胞 退变模型 WNT/Β-CATENIN信号通路 透骨消痛胶囊 补肾柔肝 活血祛风 国家自然科学基金 

分 类 号：R318[医药卫生—生物医学工程]