[1]李亚兰,黄 智,何冬凌,等.银环蛇伤中毒巴马小香猪模型构建与评估[J].陕西医学杂志,2021,50(12):1472-1476.[doi:DOI:10.3969/j.issn.1000-7377.2021.12.002]
　LI Yalan,HUANG Zhi,HE Dongling,et al.Construction and identification for Bungarus multicinctus snakebite poisoning model in Bama minipig[J].,2021,50(12):1472-1476.[doi:DOI:10.3969/j.issn.1000-7377.2021.12.002]
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银环蛇伤中毒巴马小香猪模型构建与评估
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《陕西医学杂志》[ISSN:1000-7377/CN:61-1281/TN]

卷:
50
期数:
2021年12期
页码:
1472-1476
栏目:
基础研究
出版日期:
2021-12-05

文章信息/Info

Title:
Construction and identification for Bungarus multicinctus snakebite poisoning model in Bama minipig
作者:
李亚兰; 黄 智; 何冬凌; 程晓阳; 莫财锋; 宋天麟; 廖 明
(广西医科大学生命科学研究院,广西 南宁 530021)
Author(s):
LI Yalan; HUANG Zhi; HE Dongling; CHENG Xiaoyang; MO Caifeng; SONG Tianlin; LIAO Ming
(Institute of Life Sciences,Guangxi Medical University,Nanning 530021,China)
关键词:
银环蛇;  巴马小香猪;  中毒;  模型;  半数致死量
Keywords:
Bungarus multicinctus;  Bama minipig;  Poisoning;  Model;  LD50
分类号:
R 996.3
DOI:
DOI:10.3969/j.issn.1000-7377.2021.12.002
文献标志码:
A
摘要:
目的:建立符合银环蛇咬伤临床中毒特点和表现的巴马小香猪模型。方法:通过银环蛇伤中毒小鼠半数致死量(LD50),按照体表面积比率换算等效剂量法得出银环蛇毒对巴马小香猪的理论LD50。参照理论LD50,选取18只健康巴马小香猪随机分为对照组以及0.12、0.09、0.06、0.03、0.01 mg/kg银环蛇毒剂量组,每组3只,分析不同剂量组巴马小香猪死亡情况,确定银环蛇伤中毒巴马小香猪的最佳造模剂量。造模后,进行动物行为学观察,抽取3、12、24 h静脉血进行血液学指标检测,并取心肌、肺、肝、肾及脑组织进行病理学观察。结果:银环蛇伤中毒巴马小香猪理论LD50为0.094 mg/kg。0.03 mg/kg组小香猪病死率为33.33%,该剂量符合毒理学以LD50造模原则,小香猪病死率与银环蛇毒临床致死率相符,因此将0.03 mg/kg作为最佳造模剂量。造模后,与对照组相比,模型组注射蛇毒后3 h后,白细胞计数(WBC)、K+浓度显著降低,丙氨酸氨基转移酶(ALT)、肌酐(Cr)、肌酸激酶(CK)及HCO3-浓度则显著增加(均P<0.05); 注射蛇毒12 h后,红细胞计数(RBC)显著增加(P<0.05); 超敏C反应蛋白(hs-CRP)则在注射蛇毒3～24 h内维持升高状态(均P<0.05)。病理学结果显示,模型组心肌、肺及脑组织均存在不同程度的损伤,肝肾组织损伤不明显。结论:以0.03 mg/kg银环蛇毒肌内注射,可以成功构建稳定性好、重复性高、符合临床特点的银环蛇咬伤中毒巴马小香猪模型。
Abstract:
Objective:To establish a Bama minipig model that was in line with the clinical characteristics and manifestations of Bungarus multicinctus snakebite poisoning.Methods:According to the body surface area ratio conversion equivalent dose method,the theoretical LD50 of Bungarus multicinctus venom to Bama minipig was obtained through LD50 of Bungarus multicinctus snakebite mice.According to the theoretical LD50,18 healthy Bama minipig were randomly divided into control group,and 0.12,0.09,0.06,0.03,0.01 mg/kg Bungarus multicinctus venom group(n=3).The death of Bama minipigs in different dose groups was analyzed,and the best dosage was determined.After modeling,animal behavior observation was performed,3-,12-,24-hours venous blood was drawn for hematological indexes detection,and the tissues of myocardium,lung,liver,kidney and brain were taken for pathological observation.Results:The theoretical LD50 of Bama minipig with Bungarus multicinctus snakebite poisoning was 0.094 mg/kg.The mortality rate of the minipig in the 0.03 mg/kg group was 33.33%.This dose complied with the toxicological principle of modeling with LD50.The mortality of the minipig was consistent with the clinical lethality of Bungarus multicinctus snakebite poisoning.Therefore,0.03 mg/kg was the best model dose.After modeling,compared with the control group,3 hours after injection of snake venom,the white blood cell(WBC)and K+ concentration of the model group were significantly reduced,alanine aminotransferase(ALT),creatinine(Cr),creatine kinase(CK)and HCO3- concentration increased significantly(all P<0.05); 12 hours after injection of snake venom,the red blood cell(RBC)increased significantly(P<0.05); the hypersensitivity C-reactive protein(hs-CRP)maintained an elevated state within 3 to 24 hours after injection with snake venom(all P<0.05).The pathological results showed that the myocardium,lungs and brain tissues of the model group had varying degrees of damage,and the damage of liver and kidney tissues were not obvious.Conclusion:Intramuscular injection of 0.03 mg/kg of Bungarus multicinctus venom can successfully construct a Bama minipig model with Bungarus multicinctus snakebite poisoning,which has good stability,high reproducibility and was in line with clinical characteristics.

参考文献/References:

[1] Gutierrez JM,Calvete JJ,Habib AG,et al.Snakebite envenoming[J].Nat Rev Dis Primers,2017,3:17063.
[2] Lin B,Zhang JR,Lu HJ,et al.Immunoreactivity and neutralization study of Chinese Bungarus multicinctus antivenin and lab-prepared anti-bungarotoxin antisera towards purified bungarotoxins and snake venoms[J].PLoS Neglected Tropical Diseases,2020,14(11):e0008873.
[3] Longbottom J,Shearer FM,Devine M,et al.Vulnerability to snakebite envenoming:A global mapping of hotspots[J].Lancet,2018,392(10148):673-684.
[4] Ye F,Zheng Y,Wang X,et al.Recognition of Bungarus multicinctus venom by a DNA aptamer against beta-bungarotoxin[J].PLoS One,2014,9(8):e105404.
[5] 刘金满,卢悟广,陈永根,等.银环蛇蛇毒中具有镇痛活性的多肽成分的生化分离和活性鉴定[J].世界科学技术-中医药现代化,2020,22(1):126-133.
[6] 邓海霞.广西银环蛇咬伤中毒147例临床观察[D].南宁:广西医科大学,2008.
[7] Mao YC,Liu PY,Chiang LC,et al.Bungarus multicinctus multicinctus snakebite in Taiwan[J].Am J Trop Med Hyg,2017,96(6):1497-1504.
[8] Utkin YN,Gantsova EA,Andreeva TV,et al.Venoms of kraits Bungarus multicinctus and Bungarus fasciatus contain anticoagulant proteins[J].Doklady Biochemistry and Biophysics,2015,460(1):53-58.
[9] 梁伟新,黄永良,朱明章,等.自制真空负压吸引装置辅助治疗毒蛇咬伤快速康复73例临床研究[J].陕西医学杂志,2019,48(3):362-365.
[10] 陈雨荣,安星兰,汪正铸,等.猪作为人类疾病模型的研究进展[J].中国比较医学杂志,2020,30(7):110-119.
[11] 庞国峰.巴马香猪的开发与饲养[J].中国动物保健,2016,18(7):21-22.
[12] Gutierrez JM,Lewin MR,Williams DJ,et al.Varespladib(LY315920)and methyl varespladib(LY333013)abrogate or delay lethality induced by presynaptically acting neurotoxic snake venoms[J].Toxins(Basel),2020,12(2):131.
[13] 巩红岩,郑 芳,胡晓慧,等.迷走神经刺激减轻内毒素致小鼠急性肺损伤时α7 nAChR信号通路与调节性T细胞的关系[J].中华麻醉学杂志,2018,38(12):1513-1516.
[14] 黄秋园,黄 超,陈荣芳,等.拟胆碱药物对金环蛇、银环蛇毒中毒小鼠的解毒活性研究[J].蛇志,2020,32(1):11-15.
[15] 赵 燕,戴 维.持续性床旁血液滤过联合灌流在毒蛇咬伤救治中的意义[J].陕西医学杂志,2017,46(12):1712-1713.
[16] 方雪红,苏晓丽,雒 娜.负压封闭治疗技术用于毒蛇咬伤的效果评价[J].陕西医学杂志,2015,44(2):228-229.
[17] 王 威,陈泉芳,巫艳彬,等.广西境内两种常见神经毒类毒蛇咬伤中毒的临床特点与院前急救策略分析[J].中国全科医学,2014,17(14):1671-1673.
[18] 周 凤,邓幼清,陈小红,等.银环蛇中毒后Lance-Adams综合征1例[J].疑难病杂志,2016,15(5):528-529.
[19] 邓海霞,李其斌.银环蛇咬伤中毒发病机制和治疗的研究进展[J].蛇志,2008,20(1):35-46.
[20] Liu Y,Chen JY,Shang HT,et al.Light microscopic,electron microscopic,and immunohistochemical comparison of Bama minipig(sus scrofa domestica)and human skin[J].Comp Med,2010,60(2):142-148.
[21] 吴丽红,田雨光,范 沛,等.小型猪在生物医药研究中的应用概述[J].中国比较医学杂志,2010,20(10):77-78.
[22] Shan LL,Gao JF,Zhang YX,et al.Proteomic characterization and comparison of venoms from two elapid snakes(Bungarus multicinctus and Naja atra)from China[J].Journal of Proteomics,2016,138(14):83-94.
[23] Liang Q,Huynh TM,Ng YZ,et al.In vitro neurotoxicity of Chinese krait(Bungarus multicinctus)venom and neutralization by antivenoms[J].Toxins(Basel),2021,13(1):49.
[24] 程明峰,丁小凤,张翠芳,等.急救护理路径在银环蛇咬伤重症患者急救护理中的应用[J].蛇志,2020,32(3):286-288.
[25] Hung HT,Hojer J,Du NT.Clinical features of 60 consecutive ICU-treated patients envenomed by Bungarus multicinctus[J].Southeast Asian J Trop Med Public Health,2009,40(3):518-524.
[26] Wang N,Wu XD,Zhang SS,et al.Local tissue necrosis and thrombocytopenia following Bungarus multicinctus envenomation in a child[J].Archivos Argentinos De Pediatria,2021,119(1):e80-e83.
[27] 龙 辉,徐自强.缺抗蛇毒血清治疗重症银环蛇咬伤的护理体会[J].蛇志,2015,27(4):405-407.

备注/Memo

备注/Memo:
基金项目:国家自然科学基金资助项目(81860344); 广西壮族自治区自然科学基金资助项目(2021GXNSFAA075025,2018GXNSFAA281087)
更新日期/Last Update: 2021-12-07