中国组织工程研究 ›› 2018, Vol. 22 ›› Issue (16): 2526-2531.doi: 10.3969/j.issn.2095-4344.0761

• 组织构建实验造模 experimental modeling in tissue construction • 上一篇    下一篇

细胞色素C与Caspase-9在激素性股骨头缺血坏死中的作用

龚瑜林1,王玉鑫2,赵振群3,李鸿斌1   

  1. 1内蒙古医科大学附属医院风湿免疫科,内蒙古自治区呼和浩特市  010059;内蒙古医科大学第二附属医院,2骨肿瘤骨科,3小儿骨科,内蒙古自治区呼和浩特市  010030
  • 收稿日期:2018-03-09 出版日期:2018-06-08 发布日期:2018-06-08
  • 通讯作者: 赵振群,硕士,副主任医师,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 共同通讯作者:李鸿斌,博士,主任医师,内蒙古医科大学附属医院风湿免疫科,内蒙古自治区呼和浩特市 010059
  • 作者简介:龚瑜林,女,1984年生,内蒙古自治区乌海市人,汉族,2011年内蒙古医科大学毕业,硕士,主治医师。 共同第一作者:王玉鑫,男,汉族,1984年生,内蒙古赤峰市人,内蒙古医科大学第二附属医院骨肿瘤科,硕士,医师。
  • 基金资助:

    国家自然科学基金项目(81760391);内蒙古自治区自然科学基金项目(2015MS08121)

Roles of cytochrome C and Caspase-9 in steroid-induced avascular necrosis of the femoral head

Gong Yu-lin1, Wang Yu-xin2, Zhao Zhen-qun3, Li Hong-bin1   

  1. 1Department of Rheumatology and Immunology, the Affiliated Hospital of Inner Mongolia Medical University, Hohhot 010059, Inner Mongolia Autonomous Region, China; 2Department of Bone Oncology, 3Department of Pediatric Orthopedics, the Second Affiliated Hospital of Inner Mongolia Medical University, Hohhot 010030, Inner Mongolia Autonomous Region, China
  • Received:2018-03-09 Online:2018-06-08 Published:2018-06-08
  • Contact: Zhao Zhen-qun, Master, Associate chief physician, Department of Pediatric Orthopedics, the Second Affiliated Hospital of Inner Mongolia Medical University, Hohhot 010030, Inner Mongolia Autonomous Region, China Corresponding author: Li Hong-bin, M.D., Chief physician, Department of Rheumatology and Immunology, the Affiliated Hospital of Inner Mongolia Medical University, Hohhot 010059, Inner Mongolia Autonomous Region, China
  • About author:Gong Yu-lin, Master, Attending physician, Department of Rheumatology and Immunology, the Affiliated Hospital of Inner Mongolia Medical University, Hohhot 010059, Inner Mongolia Autonomous Region, China Wang Yu-xin, Master, Physician, Department of Bone Oncology, the Second Affiliated Hospital of Inner Mongolia Medical University, Hohhot 010030, Inner Mongolia Autonomous Region, China Gong Yu-lin and Wang Yu-xin contributed equally to this work.
  • Supported by:

    the National Natural Science Foundation of China, No. 81760391; the Natural Science Foundation of Inner Mongolia Autonomous Region, No. 2015MS08121

摘要:

文章快速阅读:

文题释义:
激素性股骨头缺血坏死:是指长期或者短期大剂量使用肾上腺皮质激素所引起的股骨头部分缺血或完全缺血,进而引起股骨头内骨细胞、骨髓造血细胞及髓腔内脂肪细胞坏死的病理过程。
TUNEL法检测细胞凋亡:细胞凋亡中染色体DNA的断裂是个渐进的分阶段的过程,DNA双链断裂或只要一条链上出现缺口而产生的一系列DNA的3’-OH末端可在脱氧核糖核苷酸末端转移酶(TdT)的作用下,将脱氧核糖核苷酸和荧光素、过氧化物酶、碱性磷酸化酶或生物素形成的衍生物标记到DNA的3’-末端,从而可进行凋亡细胞的检测。
摘要
背景:
近期研究认为激素性股骨头缺血坏死可能与骨细胞凋亡存在关联,但其确切调控机制仍不清楚,细胞色素C与Caspase-9是线粒体凋亡信号通路的关键因子,二者在激素性骨坏死的作用有待于研究证实。
目的:探讨细胞色素C与Caspase-9在激素性股骨头缺血坏死中的作用。
方法:将新西兰大耳白兔20只随机分为2组,实验组予臀肌注射甲强龙,按7.5 mg/kg注射,每周2次,对照组于相同部位注射等量生理盐水,每组10只。干预8周后采用空气栓塞法处死兔并在相对无菌条件下切取双侧股骨头标本,行组织学、透射电镜、TUNEL法检测凋亡细胞,及免疫组织化学法进行相关因子细胞色素C及Caspase-9表达的检测。
结果与结论:①细胞形态及细胞凋亡:干预8周后,与对照组比较,实验组股骨头骨组织疏松,骨小梁变细稀疏,髓腔内脂肪细胞增多,空骨陷窝百分比增高(P < 0.05),凋亡小体形成,细胞凋亡率增高(P < 0.05),细胞色素C、Caspase-9阳性表达率明显升高(P < 0.01);②相关性分析:实验组细胞凋亡率与空骨陷窝率、细胞色素C及Caspase-9阳性表达率均成明显正相关(P < 0.01);③结果说明:细胞色素C异常释放和Caspase-9高表达均与激素性股骨头缺血坏死骨细胞凋亡有关。

中国组织工程研究杂志出版内容重点:组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松组织工程
ORCID: 0000-0002-9175-1736(龚瑜林)

关键词: 股骨头, 骨坏死, 糖皮质激素, 凋亡, 细胞色素C, 骨细胞, Caspase-9, 动物模型, 发病机制

Abstract:

BACKGROUND: Steroid-induced avascular necrosis of the femoral head (SANFH) may be associated with apoptosis of osteocytes, but its regulatory mechanism remains unclear. Cytochrome C and Caspase-9 are key factors in the mitochondrial apoptosis signaling pathway, and their roles in SANFH still need further investigation.
OBJECTIVE: To investigate the roles of cytochrome C and Caspase-9 in the SANFH.
METHODS: Twenty New Zealand white rabbits were randomly divided into experimental and control groups (n=10 per group). Rabbits in the experimental group were injected with 7.5 mg/kg methylprednisolone, twice weekly, and those controls were subjected to the injection of same amount of normal saline. The treatment period was 8 weeks in both group.The rabbits were then sacrificed by air embolism to remove the bilateral femoral head specimens for histological and transmission electron microscope observations. The apoptotic cells were detected by TUNEL method and the expression levels of cytochrome C and Caspase 9 were detected by immunohistochemistry.
RESULTS AND CONCLUSION: Eight weeks after intervention, compared with the control group, the experiment group had loose femoral head, thinned trabecular bone, increased adipocytes in the medullary cavity, and increased percentage of empty lacunae (P < 0.05); the apoptotic bodies were formed, and the apoptosis rate was increased (Р < 0.05); and the positive expression rates of cytochrome C and Caspase-9 were significantly increased (P < 0.01). Correlation analysis results showed that the apoptosis rate was positively correlated with the empty lacunae rate and the positive expression rates of cytochrome C and Caspase-9 (P < 0.01). These findings indicate that the abnormal release of cytochrome C and high expression of Caspase-9 are related to the apoptosis of osteocytes in SANFH.

中国组织工程研究杂志出版内容重点:组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松组织工程

Key words: Hormones, Femur Head Necrosis, Apoptosis

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