多次眼镜蛇毒注射对家兔蛇伤中毒的保护作用  

The protective effect of multiple injections of Naja naja atra venom on rabbits with snake injury and poisoning*

作　　者：陶慧娟 宋天麟 张学荣 廖明 班建东 TAO Hui-juan;SONG Tian-lin;ZHANG Xue-rong;LIAO Ming;BAN Jian-dong(Life Sciences Institute of Guangxi Medical University,Nanning City,Guangxi,530021,China)

机构地区：[1]广西医科大学生命科学研究院,广西南宁530021

出　　处：《蛇志》2020年第1期6-10,25,共6页Journal of Snake

基　　金：国家自然科学基金资助项目(No.81360078,81860344)。

摘　　要：目的探究多次注射中华眼镜蛇毒后,家兔体内产生抗体的情况,观察多次蛇毒免疫对家兔在致死量蛇毒攻击下的保护作用。方法将36只家兔随机分为6组,其中蛇毒组和细胞毒素组分别用眼镜蛇毒及其分离出的细胞毒素于皮下注射家兔后腿,蛇毒+佐剂组和细胞毒素+佐剂组分别用经弗氏佐剂乳化的眼镜蛇毒和细胞毒素以背部多点皮下注射方式免疫家兔,蛇毒空白组和细胞毒素空白组经家兔后腿皮下注射生理盐水。各组均按0、14、28、35天免疫4次,利用双向免疫扩散和间接ELISA法检测各组家兔体内抗体水平;用致死量眼镜蛇毒、细胞毒素皮下注射对应组家兔,观察呼吸、瞳孔、精神状态等生命体征及其局部组织损伤程度、死亡时间。结果蛇毒组双向扩散为1∶4,间接ELISA为1∶64000;蛇毒+佐剂组双向扩散为1∶16,间接ELISA为1∶256000;细胞毒素组双向扩散未见明显沉淀线,间接ELISA<1∶1000;细胞毒素+佐剂组双向扩散为1∶8,间接ELISA为1∶128000。致死量蛇毒、毒素攻击后,蛇毒组家兔出现呼吸困难、四肢无力、瞳孔散大、精神萎靡,注射部位有紫色瘀斑,8 h内全部死亡,与蛇毒空白组相比,家兔死亡时间相对延长;细胞毒素组和细胞毒素空白组家兔出现口鼻大量白色泡沫样黏液,呼吸困难,但注射部位瘀紫不明显,均在5 h内全部死亡;弗氏佐剂免疫组家兔无死亡,注射部位出现皮肤损伤坏死。结论在没有佐剂的情况下多次以同种蛇毒、毒素免疫的家兔产生抗体量少,对致死量蛇毒、毒素攻击无明显保护作用,虽有一定量的抗体也无法阻止毒液中细胞毒对皮肤组织的损伤。Objective To explore the production of snake venom antibodies in rabbits after multiple injections of Naja naja atra(N.atra,Chinese cobra)venom,and to observe the protective effect of multiple injections of snake venom immunization on rabbits under lethal snake venom attack.Methods Thirty six rabbits were randomly divided into six groups.The snake venom group and the cytotoxin group were injected subcutaneously into the hind legs of rabbits with cobra venom and its separated cytotoxin.The rabbits in snake venom+adjuvant group and cytotoxin+adjuvant group were immunized with cobra venom and cytotoxin emulsified by Freund's adjuvant by multiple subcutaneous injection on the back.Normal saline was injected subcutaneously into the hind legs of rabbits in snake venom blank group and cytotoxin blank group.Each group was immunized for four times in 0,14,28 and 35 days,and then blood was taken to separate the serum.Detection of antivenom and antitoxin antibody levels in rabbits of each group by double immunodiffusion and indirect ELISA.After the lethal dose of cobra venom and cytotoxin were injected subcutaneously into the rabbits of the corresponding group,the vital signs such as breath,pupil and mental state,the degree of local tissue damage and the time of death were observed.Results The double immunodiffusion test of snake venom group was 1∶4,while the indirect ELISA method was 1∶64000.The double immunodiffusion test of snake venom+adjuvant group was 1∶16,while the indirect ELISA method was 1∶256000.There was no obvious immune precipitin line in the double immunodiffusion test of cytotoxin group,the indirect ELISA method was less than 1∶1000;the double immunodiffusion test of cytotoxin+adjuvant group was 1∶8,and the indirect ELISA method was 1∶128000.The rabbits in the snake venom group suffered from dyspnea,weakness of limbs,mydriasis and mental malaise after the lethal dose of snake venom and toxin attack.There were purple ecchymosis at the injection site,and all died within 8~10 hours.Compared with t

关 键 词：蛇咬伤 中毒 眼镜蛇毒 细胞毒素 质谱 

分 类 号：R996.3[医药卫生—毒理学]