葡萄籽原花青素拮抗激素性股骨头坏死骨细胞凋亡的作用  

Grape seed proanthocyanidins antagonize osteocyte apoptosis due to steroid-induced osteonecrosis of the femoral head

作　　者：张志峰[1] 魏晶[2] 孙韬[3] 宋津玲[4] 赵振群[5] 黄健[1] Zhang Zhifeng;Wei Jing;Sun Tao;Song Jinling;Zhao Zhenqun;Huang Jian(Department of Joint Surgery,Second Affiliated Hospital of Inner Mongolia Medical University,Hohhot 010030,Inner Mongolia Autonomous Region,China;First Department of Hand and Foot Micro-surgery,Second Affiliated Hospital of Inner Mongolia Medical University,Hohhot 010030,Inner Mongolia Autonomous Region,China;Department of Emergency,Second Affiliated Hospital of Inner Mongolia Medical University,Hohhot 010030,Inner Mongolia Autonomous Region,China;Department of Imaging,Second Affiliated Hospital of Inner Mongolia Medical University,Hohhot 010030,Inner Mongolia Autonomous Region,China;Department of Pediatric Orthopedics,Second Affiliated Hospital of Inner Mongolia Medical University,Hohhot 010030,Inner Mongolia Autonomous Region,China)

机构地区：[1]内蒙古医科大学第二附属医院关节外科,内蒙古自治区呼和浩特市010030 [2]内蒙古医科大学第二附属医院手足显微Ⅰ科,内蒙古自治区呼和浩特市010030 [3]内蒙古医科大学第二附属医院急诊科,内蒙古自治区呼和浩特市010030 [4]内蒙古医科大学第二附属医院影像科,内蒙古自治区呼和浩特市010030 [5]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》2020年第26期4146-4151,共6页Chinese Journal of Tissue Engineering Research

基　　金：内蒙古自然科学基金项目(2016MS08130),项目负责人:张志峰。

摘　　要：背景:激素性股骨头缺血坏死具有复杂的生物学过程,其发病机制至今未明,临床上缺乏行之有效的治疗手段。因此,探明激素性股骨头缺血坏死的病因学机制,仍然是该领域研究的重要内容。目的:观察葡萄籽原花青素提取物对激素性股骨头坏死骨细胞凋亡的影响。方法:日本大耳白兔27只,随机分为3组:模型组静脉注射大肠杆菌内毒素,共2次,每次100μg/kg,间隔24 h,在第2次注射完成后随即肌肉注射甲基强的松龙20 mg/kg,共注射3次,每次间隔24 h;治疗组进行相同造模,在甲基强的松龙注射同时肌肉注射葡萄籽原花青素提取物200μg/kg,共注射3次,每次间隔24 h;对照组采用相同剂量注射生理盐水;最后一次注射后第4周空气栓塞法处死动物,取双侧股骨头常规固定脱钙、包埋,切片、苏木精-伊红染色,光镜下计数空缺骨陷窝,Hoechst染色法观察细胞凋亡情况,免疫组织化学染色法观察股骨头组织Caspase-9、Bcl-2的表达。结果与结论:(1)模型组空缺骨陷窝率显著高于对照组(P<0.01);治疗组空缺骨陷窝率显著低于模型组(P<0.05),但仍高于对照组(P<0.01);(2)对照组股骨头组织凋亡细胞较少;模型组股骨头组织中存在大量凋亡细胞,细胞核呈致密浓染;治疗组中凋亡细胞数有所减少,但仍高于对照组水平;(3)模型组Caspase-9的平均吸光度显著高于对照组(P<0.01),治疗组Caspase-9的平均吸光度显著低于模型组(P<0.01),与对照组相比无显著差异;模型组Bcl-2的平均吸光度值显著低于对照组(P<0.05),治疗组Bcl-2的平均吸光度显著高于模型组(P<0.01);(4)结果表明,骨细胞凋亡参与了激素性股骨头坏死的发生发展,葡萄籽原花青素可能通过促进Bcl-2的表达和抑制Caspase-9的表达来减少骨细胞凋亡的发生。BACKGROUND:Steroid-induced avascular necrosis of the femoral head has a complex biological process,and its pathogenesis is unknown.To date,there is no effective treatment in clinical practice.Therefore,exploring the etiology of steroid-induced avascular necrosis of the femoral head is still an important content of research in this field.OBJECTIVE:To explore the effect of grape seed proanthocyanidins on osteocyte apoptosis due to steroid-induced osteonecrosis of the femoral head.METHODS:Twenty-seven Japanese white rabbits were randomly divided into three groups.The model group was intravenously injected with E.coli endotoxin,100μg/kg,twice at an interval of 24 hours.Two injections of E.coli endotoxin were followed by intramuscular injection of methylprednisolone 20 mg/kg,for 3 times.The interval was 24 hours.The treatment group was intravenously injected with E.coli endotoxin,100μg/kg,twice at an interval of 24 hours.After the second injection of E.coli endotoxin,methylprednisolone 20 mg/kg and grape seed proanthocyanidin extract 200μg/kg were injected intramuscularly three times at an interval of 24 hours.The control group was treated with the same dose of saline intravenously.The animals were killed by air embolization at the 4 th week after the last injection.Under the relatively aseptic condition,the bilateral femoral heads were routinely fixed,decalcified,embedded and sliced.Histomorphological observation and hematoxylin-eosin staining were performed to count empty bone lacunae under light microscope.Hoechst staining was used to detect cell apoptosis.The expressions of Caspase-9 and Bcl-2 in the femoral head were detected by immunohistochemical staining.RESULTS AND CONCLUSION:The percentage of empty bone lacunae in the model group was significantly higher than that in the control group(P<0.01).The percentage of empty bone lacunae in the treatment group was significantly lower than that in the model group(P<0.05),but still higher than that in the control group(P<0.01).There were few apoptotic cells in the f

关 键 词：糖皮质激素 股骨头坏死 细胞凋亡 葡萄籽原花青素 CASPASE-9 Bcl-2 

分 类 号：R446[医药卫生—诊断学] R318[医药卫生—临床医学]