自噬基因微管相关蛋白1轻链3参与模型兔激素性股骨头缺血坏死的发生与发展  

Microtubule-associated protein 1 light chain 3 is involved in the occurrence and development of steroid-induced avascular necrosis of the femoral head in a rabbit model

作　　者：马海龙 赵振群[2] 刘万林[2] 孙军[1] Ma Hailong;Zhao Zhenqun;Liu Wanlin;Sun Jun(Department of Pediatric Orthopedics,Anhui Provincial Children’s Hospital,Children’s Hospital of Anhui Medical University,the Fifth Clinical Medical College of Anhui Medical University,Hefei 230051,Anhui Province,China;Department of Pediatric Orthopedics,the Second Affiliated Hospital of Inner Mongolia Medical University,Hohhot 010030,Inner Mongolia Autonomous Region,China)

机构地区：[1]安徽医科大学第五临床医学院,安徽医科大学附属省儿童医院骨科,安徽省儿童医院骨科,安徽省合肥市230051 [2]内蒙古医科大学第二附属医院小儿骨科,内蒙古自治区呼和浩特市010030

出　　处：《中国组织工程研究》2022年第32期5167-5172,共6页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金(81360273),项目负责人:刘万林;国家自然科学基金(81460331),项目参与人:赵振群、刘万林;国家自然科学基金(81560349),项目负责人:刘万林;国家自然科学基金(81760391),项目负责人:赵振群。

摘　　要：背景:激素性股骨头缺血坏死至今仍是很棘手的难题,关于其发病机制的研究仍然具有重要意义。目的:探讨在兔激素性股骨头缺血坏死模型中,自噬基因微管相关蛋白1轻链3所发挥的作用及3-甲基腺嘌呤对其的影响。方法:健康5月龄新西兰大耳白兔36只,随机分成3组,每组12只。对照组单纯肌肉注射生理盐水,2 mL/只,每周1次,共4次;股骨头坏死组肌肉注射甲基强的松龙4 mg/kg,每周1次,共4次;3-甲基腺嘌呤组肌肉注射甲基强的松龙4 mg/kg,之后随即肌肉注射3-甲基腺嘌呤2μL/只,每周1次,共4次。实验第1,2,3,4周,各组每次分别处死3只兔,取双侧后肢股骨头。光镜下计数各组空缺骨陷窝;实时荧光定量PCR检测各组细胞内微管相关蛋白1轻链3基因的表达,Western blot鉴定各组微管相关蛋白1轻链3蛋白表达。结果与结论:①光镜观察结果显示:与对照组相比,股骨头坏死组和3-甲基腺嘌呤组标本空缺骨陷窝率均显著升高(P <0.05),但3-甲基腺嘌呤组上升率低于股骨头坏死组(P <0.05);②股骨头坏死组的微管相关蛋白1轻链3 mRNA呈高表达,同时表达量较3-甲基腺嘌呤组高(P <0.05),表示细胞自噬激活,3-甲基腺嘌呤对这一过程起到抑制作用;③结果检测到模型中微管相关蛋白1轻链3的表达,证实了细胞自噬在激素性股骨头缺血坏死发病过程中发挥了一定作用;3-甲基腺嘌呤抑制了微管相关蛋白1轻链3的表达,可以抑制激素性股骨头缺血坏死的发生及发展。BACKGROUND:Steroid-induced avascular necrosis of the femoral head is still a difficult problem,and it is of great significance to study its pathogenesis.OBJECTIVE:To discuss the role of autophagy gene microtubule-associated protein 1 light chain 3(MAP1 LC3) in ste roid-induced avascular necrosis of the femoral head and the effect of 3-methyladenine on MAP1 LC3.METHODS:A total of 36 healthy New Zealand white rabbits,aged 5 months,we re randomly divided into 3 groups,with 12 rabbits in each group.Control group was given intramuscular injection of normal saline,2 mL per rabbit,once a week,four times in total.Model group was given intramuscular injection of 4 mg/kg methylprednisolone,once a week,four times in total.3-Methyladenine group was given intramuscular injection of 4 mg/kg methylprednisolone followed by intramuscular injection of 3-methyladenine,2 μL per rabbit,once a wee k,four times in total.Three animals from each group were sacrificed at 1,2,3,and 4 weeks for sampling.The femoral heads of the bilateral hindlimbs were taken.Empty bone lacunae were counted under optical microscope.mRNA and protein expression levels of MAP1 LC3 were detected by RT-PCR and western blot assay,respectively.RESULTS AND CONCLUSION:Compared with the control group,the rate of empty lacunae increased significantly in the model and 3-methyladenine groups(P <0.05),but the rate of empty lacunae was significantly lower in the 3-methyladenine group than the model group(P <0.05).The expression of MAP1 LC3 mRNA in the model group was higher than that in the 3-methyladenine group(P <0.05),indicating the activation of autophagy is inhibited by 3-methyladenine.These findings indicate that MAP1 LC3 is expressed in the animal model,confirming that autophagy plays a ce rtain role in the pathogenesis of ste roid-induced avascular necrosis of the femoral head;3-methyladenine inhibits the expression of MAP1 LC3 and reduces the occurrence and development of ste roid-induced avascular necrosis of the femoral head.

关 键 词：激素性股骨头缺血坏死 自噬 微管相关蛋白1轻链3 3-甲基腺嘌呤 动物模型 

分 类 号：R459.9[医药卫生—治疗学] R318[医药卫生—临床医学]