半夏及其毒针晶刺激性毒性的稳定性研究

Irritant Stability of Raphides and Tubers from Pinellia ternate

目的:考察半夏毒针晶刺激性毒性的稳定性及针晶蛋白对针晶刺激性毒性的贡献,为进一步揭示半夏毒针晶产生刺激性毒性的机制提供依据。方法:不同溶剂及混合蛋白酶液浸泡处理半夏及其毒针晶,用家兔眼结膜刺激性模型考察半夏及其所含针晶的刺激性变化,用电镜扫描法观察毒针晶晶体结构变化,采用垂直凝胶电泳(SDS-PAGE)试验分析蛋白酶液浸泡前后针晶蛋白的条带变化。结果:半夏毒针晶在甲醇、乙醇浸泡24 h内刺激性逐渐消失,半夏生品在75%乙醇溶液润制2 d后刺激性降低,扫描电镜显示经过甲醇、乙醇处理的毒针晶变得易于断碎,针尖锈蚀。混合蛋白酶液浸泡3 min后的半夏毒针晶刺激性急剧下降,SDS-PAGE图谱显示蛋白酶液浸泡后的针晶蛋白被酶解。结论:半夏针晶蛋白对针晶刺激性有贡献。甲醇、乙醇浸泡可使半夏毒针晶刺激性下降,提示可能是由于极性有机溶剂作用于针晶蛋白,逐渐破坏针晶蛋白的功能。

Objective： To investigate the irritation stability of raphides from Pinellia ternata and the contribution of raphides proteins on irritation.Methods： The irritation of raphides and tubers from P.ternata treated with different solvents or protease digestion were evaluated by the Draize test.The shape and appearance of raphides treated with immersion in different solvents were showed by scanning electron microscopy,and protein bands from raphides before and after protease digestion were showed by sodium dodecyl sulfate polyacrylamide gel electrophoresis（SDS-PAGE）.Results： The raphides gradually lost the irritation when immersed in methanol and ethanol,while scanning electron micrograph showed the fragility of the methanol and ethanol treated raphides.The crude tubers of P.ternata immersed in 75% solution of ethanol also lost the acridity.When treated with protease digestion,raphides lost the irritation as well as the many protein bands on the SDS-PAGE gel gradually disappeared.Conclusion： Protein of the raphides could be involved in the raphides irritation.