通络生骨胶囊对激素性股骨头坏死模型大鼠Toll样受体4信号通路的影响  

Effect of Tongluo Shenggu Capsule on toll-like receptor 4 signaling pathway in rats with steroidinduced osteonecrosis of the femoral head

作　　者：刘金富 曾平[2] 农焦 范思奇 钱晓芬 冯程钦 雷志强 罗钧 黄佳兴 Liu Jinfu;Zeng Ping;Nong Jiao;Fan Siqi;Qian Xiaofen;Feng Chengqin;Lei Zhiqiang;Luo Jun;Huang Jiaxing(Graduate School,Guangxi University of Chinese Medicine,Nanning 530299,Guangxi Zhuang Autonomous Region,China;First Affiliated Hospital,Guangxi University of Chinese Medicine,Nanning 530023,Guangxi Zhuang Autonomous Region,China)

机构地区：[1]广西中医药大学研究生学院,广西壮族自治区南宁市530299 [2]广西中医药大学第一附属医院,广西壮族自治区南宁市530023

出　　处：《中国组织工程研究》2021年第26期4150-4155,共6页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金项目(81960876),项目负责人:曾平;广西地区自然科学基金项目(2017GXNSFAA198296),项目负责人:曾平;大数据库时代广西股骨头坏死临床数据库的建设(2018XK073),项目负责人:曾平。

摘　　要：背景:前期药理学研究发现通络生骨胶囊修复激素性股骨头坏死的机制多样,但对Toll样受体4(Toll-link receptors 4,TLR4)信号通路的影响未见报道。目的:基于TLR4信号通路探讨通络生骨胶囊治疗激素性股骨头坏死的分子机制。方法:将60只雌性SD大鼠随机分为正常组20只和造模组40只,采用内毒素联合甲强龙建立大鼠激素性股骨头坏死(SONFH)模型,正常组注射等体积生理盐水;造模完成后随机抽取正常组5只、造模组10只在光镜下观察造模效果。将造模成功的30只大鼠随机平均分为2组,分别为造模组和干预组,每组15只。干预组予通络生骨胶囊水溶液0.4 g/kg每天灌胃1次,连续4周;激素组、正常组予等剂量生理盐水每天1次灌胃4周。干预结束后取各组大鼠两侧股骨头,观察股骨头外形,进行病理学组织检测,采用Western Blot检测各组大鼠股骨头TLR4、MyD88、肿瘤坏死因子α和核因子κB蛋白表达。实验方案经广西中医药大学动物实验伦理委员会批准。结果与结论:①正常组大鼠股骨头可见外观润滑有光泽,无变形塌陷;激素组大鼠股骨头无光泽,呈紫色,面积较大;干预组大鼠股骨头趋于正常;②苏木精-伊红染色结果发现,激素组股骨头骨小梁稀疏甚至断裂骨细胞核固缩溶解、消失,空骨陷窝数量最多,与正常组和干预组相比空骨陷窝率差异有显著性意义(P<0.05);干预组可见骨小梁粗大、规则,细胞形态大多正常,较激素组有改善;③Western Blot检测结果显示,与激素组相比,正常组、干预组核因子κB、肿瘤坏死因子α、MyD88、TLR4蛋白表达量均明显降低(P<0.05);干预组与正常组TLR4和MyD88的表达量差异无显著性意义(P>0.05);④结果说明,通络生骨胶囊治疗激素性股骨头坏死的机制可能与抑制TLR4信号通路MyD88依赖型传递途径有关。BACKGROUND:Early pharmacological studies have found that Tongluo Shenggu Capsule has various mechanisms for repairing steroid-induced osteonecrosis of the femoral head(SANFH),but its influence on the toll-like receptor 4(TLR4)signaling pathway has not been documented.OBJECTIVE:To investigate the molecular mechanism of Tongluo Shenggu Capsule in SANFH treatment based on the TLR4 signaling pathway.METHODS:A total of 60 female Sprague-Dawley rats were randomly divided into a normal group(20 rats)and a model group(40 rats).Endotoxin combined with methylprednisolone was used to establish the SANFH rat model.The normal group was injected with normal saline at the same volume.After modeling,5 rats from the normal group and 10 rats from the model group were selected for observation under light microscope.The 30 rats successfully modeled were randomly divided into two groups:a hormone group and an intervention group with 15 rats in each group.The intervention group was given Tongluo Shenggu Capsule solution 0.4 g/kg once a day for 4 weeks.The hormone group and the normal group were given the same dose of normal saline for 4 weeks,once a day.The hormone and normal groups were given the same amount of normal saline once a day for 4 weeks.After the end of intervention,bilateral femoral heads of rats in each group were taken for morphological observation and pathological detection.Expressions of TLR4,MyD88,tumor necrosis factor-βand nuclear factor-κB proteins were detected by western blot.RESULTS AND CONCLUSION:The femoral head of rats in the normal group had a lustrous and smooth appearance without deformation or collapse.The femoral head of rats in the hormone group had a dull,purple appearance and a large area,while the femoral head of rats in the intervention group tended to be normal.Hematoxylin-eosin staining results showed that in the hormone group,trabeculae of the femoral head were sparse and even broken,bone nuclei were solidified,dissolved and disappeared,and the number of empty bone lacunae was the largest.Compa

关 键 词：骨 股骨头坏死 激素性股骨头坏死 通络生骨胶囊 TLR4 信号通路 大鼠 

分 类 号：R446[医药卫生—诊断学] R496[医药卫生—临床医学]