摘要
目的观察家兔激素性股骨头缺血坏死模型中自噬相关基因Beclin 1和微管相关蛋白1轻链3(microtubule-associated protein 1light chain 3,MAP1-LC3)的表达情况,探讨自噬在激素性股骨头缺血坏死中的作用。方法成年日本大耳白兔40只,随机分为实验组(n=28)与对照组(n=12)。实验组肌肉注射甲基强的松龙7.5mg/kg,2次/周,共8周;对照组肌肉注射生理盐水2mL/只,2次/周,共8周。8周后处死家兔,取双侧股骨头。左侧股骨头制成普通病理切片后进行HE染色,用免疫组化法检测Beclin 1和MAP1-LC3的表达;右侧股骨头迅速置于液氮中保存,蛋白提取后,采用Western blot方法检测股骨头中Beclin 1及MAP1-LC3蛋白的表达。结果 HE染色显示家兔激素性股骨头坏死模型制造成功。免疫组化:实验组Beclin 1和MAP1-LC3的阳性率分别为96.43%、92.86%,明显高于对照组16.67%、8.33%,差异具有统计学意义(P<0.05)。Western blot检测:实验组与对照组比较Beclin 1及MAP1-LC3蛋白表达增高,差异具有统计学意义(P<0.05)。结论肾上腺皮质激素造成的家兔早期股骨头缺血坏死模型中,自噬基因Beclin 1和MAP1-LC3的表达增高,表明自噬参与了激素性股骨头缺血坏死的发病。
Objective To investigate the expression of autophagy-related gene Beclin 1and microtubule-associated protein 1light chain 3（MAP1-LC3）in avascular necrosis of the femoral head model induced by glucocorticoids,and explore the role of autophagy in avascular necrosis of the femoral head model induced by glucocorticoids.Methods Forty mature Japanese rabbits were randomly divided into experimental group（n=28）and control group（n=12）.The experimental group was given an intramscular injection of 7.5 mg/kg of methylprednisolone twice per week for 8weeks in all;the control group was given equal isotonic Na chloride.Rabbits of each group were killed at the 8th week respectively,and the hibateral heads of femur were taken out.The left heads of femur were made into general sections,dyed by HE.The expression of Beclin-1and MAP1-LC3 were detected by immunohistochemical method.The right heads of femur were put into liquid nitrogen quickly,protein were extracted from bone tissue,right head of femur was detected the protein expression of Beclin 1and MAP1-LC3 by Western blot.Results Hematoxylin and eosin stain showed the successfuluccessful production of steroid induced avascular necrosis of the femoral head in rabbits.The expression of Beclin l and MAPI-LC3 in the experimental group were 96.43%and 92.86% respectively,and the expression of Beclinl and MAPI-LC3 in the control group were 16.67%and 8.33% respectively.And the difference was significant（P〈0.05）.Compared to the control group,the protein expression of Beclinl and MAPI-LC3 increased remarkably in the experimental group,the difference was significant（P〈0.05）.Conclusion Adrenal cortex hormones cause the rabbits early avascular necrosis of the femoral head,Beclin 1and MAP1-LC3 expression increased.This study suggests that autophagy may play an important role in the development of femoral head model induced by glucocorticoids.
出处
《实用骨科杂志》
2017年第9期811-815,共5页
Journal of Practical Orthopaedics
基金
国家自然科学基金(81360273)
