中国组织工程研究

中国组织工程研究 ›› 2021, Vol. 25 ›› Issue (11): 1688-1692.doi: 10.3969/j.issn.2095-4344.3087

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

竹叶青蛇咬伤广西巴马小香猪动物模型的构建与评价

莫财锋，程晓阳，廖  明，何冬凌，黄  智   

  1. 广西医科大学生命科学研究院，广西壮族自治区南宁市   530021
  • 收稿日期:2020-07-01 修回日期:2020-07-04 接受日期:2020-08-05 出版日期:2021-04-18 发布日期:2020-12-21
  • 通讯作者: 廖明，博士，教授，广西医科大学生命科学研究院，广西壮族自治区南宁市 530021
  • 作者简介:莫财锋，男，1987年生，广西壮族自治区博白县人，汉族，广西医科大学在读硕士，主要从事蛇毒基础研究。
  • 基金资助:
    国家自然科学基金(81860344)，项目负责人：廖明；广西自然科学基金(2018GXNSFAA281087)，项目负责人：廖明

A Guangxi Bama minipig model of Trimeresurus stejnegeri snakebite: modeling and evaluation

Mo Caifeng, Cheng Xiaoyang, Liao Ming, He Dongling, Huang Zhi   

  1. Institute of Life Sciences, Guangxi Medical University, Nanning 530021, Guangxi Zhuang Autonomous Region, China
  • Received:2020-07-01 Revised:2020-07-04 Accepted:2020-08-05 Online:2021-04-18 Published:2020-12-21
  • Contact: Liao Ming, MD, Professor, Institute of Life Sciences, Guangxi Medical University, Nanning 530021, Guangxi Zhuang Autonomous Region, China
  • About author:Mo Caifeng, Master candidate, Institute of Life Sciences, Guangxi Medical University, Nanning 530021, Guangxi Zhuang Autonomous Region, China
  • Supported by:
    the National Natural Science Foundation of China, No. 81860344 (to LM); the Guangxi Natural Science Foundation, No. 2018GXNSFAA281087 (to LM)

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摘要：

文题释义：
LD50：半数致死量(median lethal dose)，表示在规定时间内，通过指定感染途径，使一定体质量或年龄的某种动物半数死亡所需最小细菌数或毒素量。在毒理学中，是描述有毒物质毒性的常用指标。
竹叶青蛇毒：毒液中主要成分为血循毒，既含凝血毒素、心脏毒素，又含血毒素、膜活性多肽等，可引起局部和内脏出血。竹叶青蛇咬伤，从临床效果观察，伤口局部会出现剧烈的灼痛，肿胀发展迅速，其典型特征为血性水泡较多见，且出现较早，一般较少出现全身症状。

背景：目前国内外对竹叶青蛇咬伤病理机制的研究方法不多，有关动物模型的构建方法较少。
目的：构建并评价竹叶青蛇咬伤广西巴马小香猪的动物模型，为竹叶青蛇咬伤的病理机制及诊治提供理想动物模型。
方法：以竹叶青蛇蛇毒对小鼠肌肉注射的LD50值为依据，通过等效剂量系数折算法和体表面积换算法，计算竹叶青蛇毒对巴马小香猪的理论半数致死量。12只巴马小香猪随机分为正常组6只和模型组6只，模型组以竹叶青蛇毒对巴马小香猪的理论半数致死量的1/3即0.643 mg/kg为注射量，0.2 mL/kg肌肉注射建立竹叶青蛇伤模型；对照组注射等量的生理盐水。注射蛇毒后观察巴马小香猪的中毒症状，在注射蛇毒前、注射蛇毒后6 h及24 h采集2组动物血液，检测血常规、凝血四项、血生化和电解质；通过苏木精-伊红染色观察心脏组织、脑组织、肺组织、肝组织、肾组织和注射部位肌肉组织病理学变化。实验方案经广西医科大学实验动物伦理委员会批准(批准号为201909013)。
结果与结论：①模型组小猪伤口处出现肿胀并伴有血水泡，患肢和周围红肿，迅速向近心端蔓延，小猪常因疼痛难忍一直走动，实验期间未见动物死亡；②与对照组相比，模型组小猪红细胞计数升高、白细胞计数升高、血小板计数降低、凝血酶原时间增加、纤维蛋白原减少、丙氨酸氨基转移酶升高、D-二聚体升高；③与对照组相比，模型组小猪肺组织毛细血管通透性增加，有充血水肿现象；注射处肌肉组织可见水肿、出血、变性坏死的改变；其他器官组织未见明显异常；④结果说明，该方法可以较好地建立竹叶青蛇咬伤小猪动物模型，能够反映竹叶青蛇咬伤的病理生理过程，具有较好的操作性及可重复性，可用于竹叶青蛇咬伤的病理生理等机制研究。

https://orcid.org/0000-0003-4601-7273 (莫财锋) 

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

关键词: 竹叶青蛇, 巴马小香猪, 动物模型, 构建, 评价

Abstract: BACKGROUND:  At present, there are not many research methods on the pathological mechanism of Trimeresurus stejnegeri snakebite, and there are few methods for constructing animal models.
OBJECTIVE: To establish and evaluate the Guangxi Bama minipig model for research on the pathological mechanism, diagnosis and treatment of Trimeresurus stejnegeri snakebite. 
METHODS: Based on the 50% lethal dose of intramuscularly injected Trimeresurus stejnegeri venom to mice, the theoretical 50% lethal venom dose for Bama minipigs was calculated by the equivalent dose coefficient conversion and reduction algorithm, and the body surface area conversion algorithm. Twelve Bama minipigs were randomly divided into a normal group (n=6) and a model group (n=6). The model group was injected intramuscularly with 0.2 mL/kg snake venom 1/3 of the theoretical 50% lethal dose (0.643 mg/kg). The control group was injected with the same amount of normal saline. After snake venom injection, the poisoning symptoms of Bama minipigs were observed. Two groups of animal blood samples were collected before, 6 hours and 24 hours after snake venom injection. Blood routine test, four coagulation items, blood biochemistry and electrolyte were detected. Histopathological changes of the heart, brain, lung, liver, and kidney as well as the injection site were observed by hematoxylin-eosin staining. The study protocol was approved by the Laboratory Animal Ethics Committee (approval No. 201909013). 
RESULTS AND CONCLUSION: There was swelling at the wound of the piglet accompanied with blood blisters after snake venom injection. The affected limbs and the surrounding area were swollen and spread rapidly to the proximal end. The piglets walked all the time because of the pain, and no animal died during the experiment. Compared with the control group pig, the model group had higher red blood cell count, white blood cell count, alanine aminotransferase, D-dimer count, longer prothrombin time, and lower fibrinogen and platelet count. Hematoxylin-eosin staining showed that, compared with the control group, capillary permeability of piglet lung tissues in the model group was increased with hyperemia and edema. Edema, bleeding, degeneration and necrosis were seen in the muscle tissue on the injection site. No obvious abnormalities in other organs and tissues were observed. To conclude, this method can be used to establish a pig model of Trimeresurus stejnegeri snakebite that can reflect the pathophysiological process of Trimeresurus stejnegeri snakebite. It has operability and repeatability that can be used to study the pathophysiological mechanism of Trimeresurus stejnegeri snakebite.


Key words: Trimeresurus stejnegeri, Bama minipig, animal model, establishment, evaluation

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