Kalam Yasmean, Isbister Geoffrey K, Mirtschin Peter, Hodgson Wayne C, Konstantakopoulos Nicki
Monash Venom Group, Department of Pharmacology, Monash University, Vic. 3800, Australia.
J Pharmacol Toxicol Methods. 2011 Mar-Apr;63(2):137-42. doi: 10.1016/j.vascn.2010.09.001. Epub 2010 Sep 16.
Acanthophis genus (i.e. death adders) and the Naja genus (i.e. cobras) belong to the family elapidae. The current study compared the in vitro cytotoxicity of venoms from four Acanthophis spp. and three Naja spp. on rat aortic smooth muscle cells, A7r5, and rat skeletal muscle cells, L6. The ability of CSL death adder antivenom and SAIMR antivenom, for Acanthophis spp. and Naja spp. venom respectively, to negate the cytotoxicity was also examined.
A cell proliferation assay was used to determine cell viability following treatment with venom in the presence or absence of antivenom. Sigmoidal growth curves were obtained, and IC(50) values were determined.
Acanthophis spp. and Naja spp. venoms produced concentration-dependent inhibition of cell proliferation in both cell lines. Naja spp. venoms were significantly more cytotoxic than the most potent Acanthophis venom (i.e. A. antarcticus) in both cell lines. Naja spp. venoms also displayed higher sensitivity in L6 cells. SAIMR antivenom significantly inhibited the cytotoxic actions of all Naja spp. venoms in both A7r5 and L6 cells. However, death adder antivenom (CSL Ltd) was unable to negate the cytotoxic effects of Acanthophis spp. venoms.
Concentrations of the predominantly cytotoxic Naja spp. venoms used were approximately three times less than the predominantly neurotoxic Acanthophis spp. venoms. SAIMR antivenom was partially effective in neutralising the effects of Naja spp. venoms. Death adder antivenom (CSL Ltd) was not effective in negating the cytotoxic effects of venom from Acanthophis spp. These results indicate that the cell-based assay is suited to the examination of cytotoxic snake venoms and may be used in conjunction with organ bath experiments to pharmacologically characterise snake venoms. Furthermore, the results suggest that the use of a skeletal muscle cell line is likely to be more clinically relevant for the examination of cytotoxic snake venoms.
棘蛇属（即死亡蝰蛇）和眼镜蛇属（即眼镜蛇）属于眼镜蛇科。本研究比较了四种棘蛇属物种和三种眼镜蛇属物种的毒液对大鼠主动脉平滑肌细胞A7r5和大鼠骨骼肌细胞L6的体外细胞毒性。同时还检测了分别针对棘蛇属物种和眼镜蛇属物种毒液的CSL死亡蝰蛇抗蛇毒血清和南非医学研究所抗蛇毒血清消除细胞毒性的能力。
采用细胞增殖试验来确定在有或没有抗蛇毒血清的情况下毒液处理后的细胞活力。获得S形生长曲线，并确定IC(50)值。
棘蛇属物种和眼镜蛇属物种的毒液在两种细胞系中均产生浓度依赖性的细胞增殖抑制作用。在两种细胞系中，眼镜蛇属物种的毒液比毒性最强的棘蛇毒液（即南极棘蛇）的细胞毒性显著更强。眼镜蛇属物种的毒液在L6细胞中也表现出更高的敏感性。南非医学研究所抗蛇毒血清在A7r5和L6细胞中均显著抑制了所有眼镜蛇属物种毒液的细胞毒性作用。然而，死亡蝰蛇抗蛇毒血清（CSL有限公司）无法消除棘蛇属物种毒液的细胞毒性作用。
所使用的主要具有细胞毒性的眼镜蛇属物种毒液的浓度比主要具有神经毒性的棘蛇属物种毒液低约三倍。南非医学研究所抗蛇毒血清在中和眼镜蛇属物种毒液的作用方面部分有效。死亡蝰蛇抗蛇毒血清（CSL有限公司）在消除棘蛇属物种毒液的细胞毒性作用方面无效。这些结果表明，基于细胞的试验适用于检测具有细胞毒性的蛇毒，并且可以与器官浴实验结合使用，以从药理学角度表征蛇毒。此外，结果表明使用骨骼肌细胞系可能在检测具有细胞毒性的蛇毒方面更具临床相关性。
