岩豆凝集素的纯化及其分子残基的化学修饰和活性关系

岩豆种子经侵取、硫酸铵分级、ＤＥＡＥ－纤维素离子交换和ＳｅｐｈａｄｅｘＧ－１００分子筛层析，获得ＦＰＬＣ上单一的凝集素样品，对此纯化的凝集素样品用Ｎ－溴代丁二酰亚胺、Ｎ－乙基顺丁烯二酰亚胺、二流苏糖醇，碘乙酸和对氯汞苯甲酸修饰其分子内的色氨酸和巯基．经计算，该凝集素分子内含有４个色氨酸和２４个流基，当色氨酸被修饰后其凝集活性全部丧失，而巯基被修饰后对其凝血活性无任何影响，表明巯基与凝集素分子的凝血活性无关，而色氨酸则是凝集活性所必需的基团．     

Evaluation of developmental toxicity and apoptotic induction of the aqueous extract of Millettia pachycarpa using zebrafish as model organism

Extensive and indiscriminate use of synthetic compounds and natural compounds obtained from plant sources have resulted in serious threats to the aquatic ecosystem and human health. Aqueous extract of the root of the plant, Milletia pachycarpa Benth, is currently used for killing fish in the state of Manipur, India. Moreover, this plant is also used as traditional medicine in this region. Although it is widely used in traditional medicine, there is limited information available regarding the adverse effects and mechanism underlying its toxicity. This study examined the effects of exposure to aqueous extract of M. pachycarpa (AEMP) on early embryonic development of zebrafish embryos and mechanisms underlying toxicity. Zebrafish embryos treated with different concentrations of the AEMP produced embryonic lethality and developmental defects. The 96-hr-LC₅₀ of AEMP was found to be 4.276 µg/mL. Further, multiple developmental abnormalities such as pericardial edema, yolk sac edema, spinal curvature, swim bladder deflation, decreased heart rate, and delayed hatching were also observed in a dose-dependent manner. Zebrafish embryo showing moderate-to-severe developmental defects following AEMP exposure cannot swim properly. Further, this study examined oxidative stress and apoptosis in embryos exposed to AEMP. Enhanced production of ROS and apoptosis was found in brain, trunk, and tail of zebrafish embryos treated with AEMP. Data suggest that oxidative stress and apoptosis are associated with AEMP-induced embryonic lethality and developmental toxicity in zebrafish embryos.     

岩豆凝集素分子中色氨酸残基的化学修饰及其荧光光谱研究

从岩豆（Ｍｉｌｌｅｔｔｉａ ｄｉｅｌｓｉａｎａ Ｈａｒｍｓ ｅｘ Ｄｉｅｌｓ）种妇中分离纯化得到的岩豆凝集素（简称ＭＤＬ）经链霉蛋白酶水解,测得其分子中含有４个色氨酸（Ｔｒｐ）残基。用Ｎ－溴代丁二酰亚胺（ＮＢＳ）对ＭＤＬ分子中的色氨酸残基进行化学修饰,在无变性剂存在下有３．６个Ｔｒｐ残基被修饰,在变性条件下可修饰３．８个Ｔｒｐ残基。修饰后ＭＤＬ活性均丧失,且甘露糖对ＭＤＬ活性雎有保护作用。表明Ｔｒ