| 应用弹性蛋白酶诱导构建新型慢性阻塞性肺疾病合并骨质疏松症的动物模型 |
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投稿时间：2019-08-20
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| 作者 | Author | 单位 | Address | E-Mail |
| 陈文祥 |
CHEN Wen-xiang |
湖州市中心医院 浙江大学湖州医院骨科, 浙江 湖州 313003 |
Department of Orthopaedics, Huzhou Central Hospital/Zhejiang University Huzhou Hospital, Huzhou 313003, Zhejiang, China |
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| 王雍立 |
WANG Yong-li |
湖州市中心医院 浙江大学湖州医院骨科, 浙江 湖州 313003 |
Department of Orthopaedics, Huzhou Central Hospital/Zhejiang University Huzhou Hospital, Huzhou 313003, Zhejiang, China |
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| 谢子昂 |
XIE Zi-ang |
浙江大学医学院附属邵逸夫医院骨科, 浙江 杭州 310020 |
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| 范顺武 |
FAN Shun-wu |
浙江大学医学院附属邵逸夫医院骨科, 浙江 杭州 310020 |
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| 蒋雪生 |
JIANG Xue-sheng |
湖州市中心医院 浙江大学湖州医院骨科, 浙江 湖州 313003 |
Department of Orthopaedics, Huzhou Central Hospital/Zhejiang University Huzhou Hospital, Huzhou 313003, Zhejiang, China |
jiangxuesheng2000@163.com |
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| 期刊信息：《中国骨伤》2020年，第33卷，第4期，第356-362页 |
| DOI：10.12200/j.issn.1003-0034.2020.04.013 |
| 基金项目:浙江省科技厅公益技术应用研究项目（编号：2017C37119） |
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中文摘要:
目的：建立并评价弹性蛋白酶诱导小鼠慢性阻塞性肺疾病（chronic obstructive pulmonary disease，COPD）合并骨质疏松症模型。
方法：24只健康雌性8周龄C57BL/6小鼠（重约18 g）随机分为3组，对照组予以气管内滴注生理盐水，实验1组和实验2组气管内滴注弹性蛋白酶，对照组和实验1组继续饲养8周后处死，实验2组继续饲养12周后处死。HE染色评估对照组和实验组小鼠肺组织与胫骨组织病理学改变。ELISA检测支气管肺泡灌洗液与血清相关炎症因子含量。Micro-CT检测小鼠股骨骨量相关参数。实时荧光定量PCR实验检测破骨与成骨相关基因表达情况。
结果：肺组织病理显示实验组肺泡结构紊乱，肺泡壁变薄或断裂，肺泡腔扩大。肺泡灌洗液炎症因子IL-6与TNF-α较对照组明显升高（P<0.001），血清炎症因子IL-1β与TNF-α较对照组明显上升（P<0.001）。实验组BV/TV （骨体积分数）、Tb.Th （平均骨小梁厚度）与Tb.N （平均骨小梁数）较对照组均明显降低（P<0.05），实验组Tb.Sp （平均骨小梁分离度）和BS/BV （骨表面积分数）较对照组明显增加（P<0.01）。与对照组相比，实验组小鼠胫骨组织破骨相关标志基因表达增加（P<0.05），成骨相关标志基因表达下降（P<0.05）。实验1组与实验2组结果均呈时间依赖。
结论：该研究应用弹性蛋白酶成功构建COPD合并骨质疏松症模型，为今后探究COPD合并骨质疏松症的发病机制提供合适的动物模型。 |
| 【关键词】弹性蛋白酶 肺疾病,慢性阻塞性 骨质疏松症 小鼠 |
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| Establishment of a novel mouse mode of elastase-induced chronic obstructive pulmonary disease related osteoporosis |
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ABSTRACT
Objective: To establish and evaluate the model of chronic obstructive pulmonary disease(COPD) with osteoporosis induced by elastase in mice.
Methods: Twenty-four healthy female 8-week-old C57BL/6 mice(weighing about 18 g) were randomly divided into three groups. The control group was given intratracheal drip of normal saline,the experimental group 1 and the experimental group 2 were given intratracheal drip of elastase,the control group and the experimental group 1 were kept for 8 weeks and then killed,the experimental group 2 was kept for 12 weeks and then killed. HE staining was used to evaluate the histopathological changes of lung and tibia in the control and experimental groups. The levels of serum inflammatory factors and broncho alveolar lavage factors(BALF) were detected by ELISA. Micro CT was used to detect the bone mass related parameters of mouse femur. The expression of osteoclastic and osteogenic genes was detected by real-time fluorescence quantitative PCR.
Results: Lung histopathology showed that the structure of alveoli in the experimental group was disordered,the walls of alveoli became thin or broken,and the alveoli cavity expanded. IL-6 and TNF-α in BALF were significantly higher than those in control group(P<0.001),while IL-1β and TNF-α in serum inflammatory factors were significantly higher than those in control group (P<0.001). BV/TV(bone volume fraction),TB.Th(average bone trabecular thickness) and TB.N(average bone trabecular number) in the experimental group were significantly lower than those in the control group(P<0.05),TB.Sp(average bone trabecular separation) and BS/BV(bone surface area fraction) in the experimental group were significantly higher than those in the control group(P<0.01). Compared with the control group,the expression of osteoclast related marker genes increased in the experimental group (P<0.05),but decreased in the experimental group(P<0.05). The results of experiment 1 and experiment 2 were time-dependent.
Conclusion: In this study,elastase was used to construct a COPD model with osteoporosis successfully,which provides a suitable animal model for the future study of the pathogenesis of COPD with osteoporosis. |
| KEY WORDS Elastase Pulmonary disease,chronic obstructive Osteoporosis Mice |
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| 引用本文，请按以下格式著录参考文献： |
| 中文格式: | 陈文祥,王雍立,谢子昂,范顺武,蒋雪生.应用弹性蛋白酶诱导构建新型慢性阻塞性肺疾病合并骨质疏松症的动物模型[J].中国骨伤,2020,33(4):356~362 |
| 英文格式: | CHEN Wen-xiang,WANG Yong-li,XIE Zi-ang,FAN Shun-wu,JIANG Xue-sheng.Establishment of a novel mouse mode of elastase-induced chronic obstructive pulmonary disease related osteoporosis[J].zhongguo gu shang / China J Orthop Trauma ,2020,33(4):356~362 |
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