β-蜕皮甾酮调控骨髓间充质干细胞成骨分化对幼兔激素性股骨头坏死的影响  

Effect of β-Ecdysterone modulation of osteogenic differentiation of bone mesenchymal stem cells on hormonal femoral head necrosis in juvenile rabbits

作　　者：何哲希 杨嘉飞 姜旭 朱兴涛 刘露 俞松 He Zhexi;Yang Jiafei;Jiang Xu;Zhu Xingtao;Liu Lu;Yu Song(Bone Injury College,Guizhou University of Traditional Chinese Medicine,Guiyang 550025,China;Department of Pediatric Surgery,Affiliated Hospital of Guizhou Medical University,Guiyang 550004,China;Department of Clinical Medicine,Zunyi Medical and Pharmaceutical College,Zunyi 563000,China)

机构地区：[1]贵州中医药大学骨伤学院,贵阳550025 [2]贵州医科大学附属医院小儿外科,贵阳550004 [3]遵义医药高等专科学校临床医学系,遵义563000

出　　处：《中华实用儿科临床杂志》2025年第3期207-213,共7页Chinese Journal of Applied Clinical Pediatrics

基　　金：贵州省高层次创新型人才项目(黔科合平台人才(2020)6015-2);贵州省科技计划项目(黔科合支撑(2021)一般076)。

摘　　要：目的观察牛膝活性成分β-蜕皮甾酮(β-Ecd)对幼兔激素性股骨头坏死的影响,并探讨其有关机制。方法动物实验研究。60只新西兰幼兔采用随机数字表法分为对照组、模型组及干预组,每组20只。模型组和干预组幼兔双侧臀肌交替注射醋酸泼尼松龙(7.5 mg/kg),2次/周;干预组首次注射醋酸泼尼松龙时开始皮下注射β-Ecd(0.5 mg/kg),5次/周;对照组和模型组注射等量生理盐水。造模8周后处死动物,取双侧股骨头行大体观察;微型计算机断层扫描(Micro-CT)观察骨小梁微结构,测量骨微结构参数;组织学染色观察骨组织形态变化;免疫组织化学检测股骨头组织中成骨、成脂相关因子的表达情况。Western blot检测股骨头组织中成骨、成脂相关蛋白的表达含量;将兔骨髓间充质干细胞(BMSCs)分为空白对照组、地塞米松组及β-Ecd干预组,地塞米松组与β-Ecd干预组用地塞米松诱导细胞损伤,β-Ecd干预组予最佳药物浓度β-Ecd干预。Western blot检测各组细胞成骨、成脂相关蛋白的表达含量;实时荧光定量反转录聚合酶链反应(RT-qPCR)测定成骨、成脂相关标志基因中mRNA的表达情况。结果实验期间9只动物死亡,最终纳入对照组19只、模型组15只、干预组17只。大体观察及Micro-CT显示,与对照组相比,模型组股骨头晦暗偏灰,出现暗红色坏死区域,骨小梁疏松、变细、排列紊乱、部分断裂;干预组的变化介于对照组和模型组之间。与对照组相比,模型组和干预组的骨密度、骨小梁数量、骨小梁厚度、骨小梁相对体积均下降,骨小梁分离度升高,但干预组较模型组更轻;组间比较差异均有统计学意义(F=12.78、45.52、32.74、64.08、8.83,均P<0.05)。病理组织学显示,对照组骨小梁排列整齐、粗壮饱满、骨细胞数量较多、偶见极个别空骨陷窝;模型组骨小梁排列稀疏、断裂、骨细胞减少,空骨陷窝增多、增大;干预组较模型组�Objective:To investigate the effect ofβ-Ecdysterone(β-Ecd),an active ingredient in cow′s knee,on hormonal femoral head necrosis in young rabbits and explore the mechanism involved.Methods:An animal study.Sixty New Zealand young rabbits were divided into control,model and intervention groups by random number table method,with 20 rabbits in each group.Prednisolone acetate(7.5 mg/kg)was injected bilaterally into the gluteal muscle of rabbits in both model and intervention groups twice a week.β-Ecd(0.5 mg/kg)was injected subcutaneously at the time of the first injection of Prednisolone acetate in the intervention group for 5 times a week.An equal amount of saline was injected into rabbits in control and model groups.Eight weeks after modelling,animals were put to death,and femoral heads were taken from both sides for gross observation.Micro computed tomography(Micro-CT)was used to analyze the microstructure of bone trabeculae and to measure bone microstructural parameters.Histological staining was used to detect changes in the morphology of bone tissues.Immunohistochemistry and Western blot were used to examine the expression of osteogenic and lipogenic factors and proteins in the femoral head tissue.Rabbit bone marrow mesenchymal stem cells(BMSCs)were divided into a blank control group,a Dexamethasone group and aβ-Ecd intervention group.Cell damage was induced by Dexamethasone in Dexamethasone group andβ-Ecd intervention group,and theβ-Ecd intervention group was given the optimal concentration ofβ-Ecd.Western blot was used to detect the expression of osteogenic and lipogenic proteins in the cells of each group.Real-time quantitative polymerase chain reaction(RT-qPCR)was used to determine the mRNA expression of osteogenic and lipogenic marker genes.Results:After excluding 9 rabbits that died during the experimental period,51 rabbits were finally included in the study,with 19 in the control group,15 in the model group and 17 in the intervention group.Gross observation and Micro-CT showed that compared with th

关 键 词：骨髓间充质干细胞 β-蜕皮甾酮 激素性股骨头坏死 动物模型 幼兔 治疗 

分 类 号：R73[医药卫生—肿瘤]