江浙蝮蛇毒镇痛组分的分离纯化及其理化性质

目的　分离筛选江浙蝮蛇毒中主要镇痛组分 ,研究其分子量、等电点、纯度、氨基酸序列、稳定性等理化性质 ,以及镇痛作用、机体耐受性和依赖性等药理学性质。方法　采用离子交换和分子筛 ,以柱层析法分离蛇毒组分。用生化测定方法 ,测定其理化性质。用热板法和醋酸扭体法等确定最佳镇痛组分 ,并用纳洛酮催促和自然戒断实验、耐受性实验考察其依赖性和耐受性。结果　分离获得 14个蛋白组分 ,经ED50 /LD50 筛选出最佳镇痛组分C4 ,鉴定结果表明纯度为电泳纯 ,HPLC相对纯度为 92 .16 % ,分子量 16 .6ku ,等电点 8.8,氨基酸序列与磷脂酶A2 同源 ,为一种新的镇痛蛋白 ,并获得其紫外吸收特征峰、镇痛耐受和依赖性等性质。结论　C4组分具有显著的镇痛效果 ,未发现耐受性和依赖性 ,值得进一步研究开发。

Purification and properties of analgesic fraction in venom of Agkistrodon halys Pallas

AIM To separate main analgesic fraction in v enom of Agkistrodon halys Pallas, study its physical and chemical p roperties such as molecular weight, isoelectric point, purity, amino acid sequen ce, stability, its pharmacological properties such as analgesic function, tolera nce, dependence. METHODS To separate snake venom by column chromato graphy with ion exchange and size exclusion, determine its properties by bioche mical methods, find the best analgesic fraction by hot plate test and writhing t est, observe its dependence and tolerance by naloxone precipitation test, natura l abstinence test and tolerance test. RESULTS Fourteen fractions we re obtained, the best analgesic C4 was chosen according to the value of ED 50/LD 50 . Test results showed it had a single blot in electropherosis, relative purity 92.16%(HPLC), M r 16.6 ku, pI 8.8, AA squence homogenous from phos pholipase A 2. Its special peaks in UV spectrum, its tolerance and depedence we re also obtained. CONCLUSION C4 fraction has significant analge si c effect, it has no dependence and tolerance. It is worth further developing.