【摘 要】
：
目的：对广西眼镜蛇毒中磷脂酶A2进行分离纯化并测定其对肝星状细胞HSC-T6的增值抑制作用.方法：采用Sephadex G-50凝胶层析柱、CM-Sepharose CL-6B离子交换柱、Macro-prep H
【出 处】
：
第12届生物毒素研究及医药应用学术大会
论文部分内容阅读
目的：对广西眼镜蛇毒中磷脂酶A2进行分离纯化并测定其对肝星状细胞HSC-T6的增值抑制作用.方法：采用Sephadex G-50凝胶层析柱、CM-Sepharose CL-6B离子交换柱、Macro-prep HighS预装柱结合的方法分离广西眼镜蛇粗毒,经平板法测定各峰的PLA2活性；经SDS-PAGE电泳鉴定终产物纯度并测定分子量,经NanoLC-ESI-MS/MS鉴定其组分；CCK-8法测定PLA2对肝星状细胞(HSC-T6)的增殖抑制作用,找出引起其凋亡的最小毒性浓度.结果：Sephadex G-50凝胶层析柱、CM-Sepharose CL-6B离子交换柱、Macro-prep HighS预装柱层析法,得到第Ⅲ峰具PLA2活性,且达到电泳纯,经NanoLC-ESI-MS/MS鉴定其为PLA2,分子量约为14.06kd；PLA2在0～1μg/ml的浓度下对HSC-T6细胞具有一定的促增殖作用,2tg/ml时细胞数达到最大值,4～16μg/ml时对细胞生长有抑制作用,且随浓度增大细胞数减小.
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