Department of Physiology, University of Calcutta, Kolkata, India.
Indian J Med Res. 2012 Jul;136(1):40-5.
BACKGROUND & OBJECTIVES: Phospholipase A2 (PLA2 ) is one of the major constituents of krait venom associated with several pathophysiological actions like myotoxicity, cardiotoxicity, neurotoxicity, etc. As there was no specific antiserum available against Bungarus fasciatus venom, this study was done with synthetic herbal compounds, anti PLA2 rabbit antiserum and commercial polyvalent snake venom antiserum to neutralize the PLA2 induced toxicities in experimental models.
B. fasciatus venom phospholipase A2 fraction 38 (BF-38) was isolated by ion exchange chromatography, molecular weight was determined by mass spectrometry and its N terminal amino acid sequence was identified. Monospecific rabbit antiserum was raised against the PLA2 in presence of Freund complete adjuvant. The neutralization of PLA2 induced toxicities was done in in vitro and in in vivo models using synthetic herbal compounds, anti PLA2 rabbit antiserum and commercial polyvalent snake venom antiserum.
A toxic PLA2 (BF-38) was purified from the B. fasciatus venom by CM-cellulose and HPLC, of 13.17 kDa and a minor band of 7.3 kDa using ESI-MS. The 13.17 kDa PLA2 sequence was NLYQFKNMIQC. The 7.3 kDa toxin sequence was RKCLTKYSQDNES and was found to be <10 per cent w/w. Anti PLA2 rabbit antiserum produced faint precipitant band in immunogel diffusion and showed low titre value. The commercial polyvalent snake venom antiserum, anti PLA2 rabbit antiserum and the synthetic herbal compounds neutralized the PLA2 induced toxicities at different intensities.
INTERPRETATION & CONCLUSIONS: Our results suggested that synthetic herbal compound (BA) along with antiserum might provide effective protection against PLA2 induced toxicities of B. fasciatus venom.
磷脂酶 A2(PLA2)是眼镜蛇科蛇毒的主要成分之一,与多种病理生理作用有关,如肌毒性、心脏毒性、神经毒性等。由于没有针对金环蛇毒的特异性抗血清,因此本研究使用合成草药化合物、抗 PLA2 兔抗血清和商业多价蛇毒抗血清来中和实验模型中 PLA2 诱导的毒性。
通过离子交换色谱法分离 B. fasciatus venom 磷脂酶 A2 分馏 38(BF-38),通过质谱法确定分子量,并鉴定其 N 末端氨基酸序列。在弗氏完全佐剂存在下针对 PLA2 产生单特异性兔抗血清。使用合成草药化合物、抗 PLA2 兔抗血清和商业多价蛇毒抗血清在体外和体内模型中中和 PLA2 诱导的毒性。
通过 CM-纤维素和 HPLC 从 B. fasciatus 毒液中纯化出一种有毒的 PLA2(BF-38),ESI-MS 显示其分子量为 13.17 kDa,还有一个 7.3 kDa 的小带。13.17 kDa PLA2 序列为 NLYQFKNMIQC。7.3 kDa 毒素序列为 RKCLTKYSQDNES,<10%w/w。抗 PLA2 兔抗血清在免疫凝胶扩散中产生微弱的沉淀带,效价较低。商业多价蛇毒抗血清、抗 PLA2 兔抗血清和合成草药化合物以不同的强度中和 PLA2 诱导的毒性。
我们的结果表明,合成草药化合物(BA)与抗血清一起可能为金环蛇毒 PLA2 诱导的毒性提供有效的保护。