Khoobdel Mehdi, Zahraei-Salehi Taghi, Nayeri-Fasaei Bahar, Khosravi Mohammad, Omidian Zahra, Motedayen Mohammad Hassan, Akbari Abolfazal
Health Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran.
Department of Microbiology, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran.
J Arthropod Borne Dis. 2013 Aug 31;7(2):139-46. eCollection 2013.
Scorpions stings are a health problem in many parts of the world. Mesobuthus eupeus (Buthidae) is the most prevalent species in the Middle East and Central Asia. Definition of toxicogenic and immunogenic characteristics of the venom is necessary to produce antidote. In this study, the noted properties of M. eupeus venom were evaluated.
Venom was obtained by milking M. eupeus scorpions for lyophilization. Toxicity was determined after injecting the venom to albino mice and calculating LD50. Polyclonal antibodies against M. eupeus venom were obtained from immunized rabbits. The CH-Sepharose 4B column was used for isolating the specific antibodies. 10 mg of the affinity-purified antibodies were conjugated with a CH-Sepharose 4B column and M. eupeus venom was applied to the column. The bound fragments were eluted using hydrogen chloride (pH: 2.5). Crude venom and affinity-purified fractions of the venom were analyzed by SDS-PAGE technique.
Lethal dose (LD) was 8.75, 11.5 and 4.5 mg/kg for IP, SC and IV respectively. The LD50 of M. eupeus venom was 6.95 mg/kg. The crude venom had 12 detectable bands with molecular weights of 140, 70, 50, 33, 30, 27, 22, 18, 14, 10 kDa and two bands less than 5 kDa. The affinity-purified venom presented eight bands. The 27 kDa band was clearly sharper than other bands but 70, 18, 10 and one of the less than 5 kDa bands were not observed.
Contrary to popular belief, which know scorpion venom as non-immunogenic composition, the current study was shown that the most fractions of the M. eupeus are immunogenic.
蝎子蜇伤是世界许多地区的一个健康问题。中东和中亚地区最常见的蝎子种类是真钳蝎（钳蝎科）。明确毒液的毒性和免疫原性特征对于生产抗毒血清至关重要。在本研究中，对真钳蝎毒液的显著特性进行了评估。
通过挤取真钳蝎毒液进行冻干获取毒液。将毒液注射到白化小鼠体内并计算半数致死量（LD50）来测定毒性。从免疫兔中获取抗真钳蝎毒液的多克隆抗体。使用CH-琼脂糖4B柱分离特异性抗体。将10mg亲和纯化抗体与CH-琼脂糖4B柱偶联，然后将真钳蝎毒液应用于该柱。用盐酸（pH：2.5）洗脱结合片段。采用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）技术分析粗毒液和亲和纯化的毒液组分。
腹腔注射（IP）、皮下注射（SC）和静脉注射（IV）的致死剂量（LD）分别为8.75、11.5和4.5mg/kg。真钳蝎毒液的LD50为6.95mg/kg。粗毒液有12条可检测条带，分子量分别为140、70、50、33、30、27、22、18、14、10kDa以及两条小于5kDa的条带。亲和纯化的毒液呈现8条带。27kDa条带明显比其他条带更清晰，但未观察到70、18、10kDa条带以及一条小于5kDa的条带。
与普遍认为蝎子毒液是非免疫原性成分的观点相反，当前研究表明真钳蝎毒液的大部分组分具有免疫原性。
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