灭螺药物对钉螺和财鱼GOT酶活力及其蛋白表达的影响研究
发布时间：2019-02-28 14:15
【摘要】：湖北钉螺是日本血吸虫的唯一中间宿主,药物灭螺能有效预控血吸虫病流行。NiC是WHO唯一指定、我国常用的化学灭螺药物,但存在水溶解度低、靶标/点的选择毒性差及价格昂贵等技术经济难点。因此,需要紧密结合预控血吸虫病的国家重大技术需求研发新型灭螺药物。本文以苯胺NiC和灭螺植物水溶性甾体皂甙/生物碱为供试药物,以钉螺和财鱼为靶标,探讨药物对其蛋白质表达和酶活力的影响,这为新型灭螺药物研发的先导物和特定靶点分选提供依据。 1、钉螺软体和财鱼肝脏GOT酶催化最适条件的研究 通过L25(56)实验及极/方差分析,测得钉螺软体/财鱼肝脏匀浆液GOT酶催化最适条件分别在酶液浓度0.2/0.4mg/mL、底物浓度5.0/5.0μmol/L、反应体系pH7.5/8.0、反应时间60/20min及反应温度45/45℃时,其转氨基速率最高。提示物种间GOT酶催化动力学不同。这为灭螺药物对其活性影响研究提供实验依据。 2、灭螺药物温孵/浸泡对钉螺和财鱼的GOT酶活力影响 2.1药物温孵离体对钉螺软体的GOT酶活力 浓度效应：梯度质量数的NiC和吉祥草、黄果茄及盾叶薯蓣水提取物,分别与酶温孵20min,在其0.25、0.5、0.5及2.0mg/L的酶活力诱导率最高,分别达到19.3、23.7、18.9及11.52%,提示钉螺GOT酶对吉祥草甾体生物碱的敏感性最高。 2.2 NiC温孵离体财鱼肝脏的GOT酶活力 浓度效应：梯度质量数的NiC分别与酶温孵30min,0.125mg/L NiC诱导率高达39.94%。 时间效应：0.25mg/L NiC与酶经时温孵,40min温孵酶活力诱导率高达44.86%。 2.3 NiC染毒财鱼肝脏的GOT酶活力 梯度质量的NiC浸泡染毒财鱼,1.5mg/L时财鱼死亡时间最短3.5h,GOT酶活力诱导率最大113.99%。 提示NiC诱导GOT酶活力可能是其致死财鱼的靶点之一 3、NiC染毒对财鱼肝脏蛋白质表达影响 0.5mg/L NiC染毒财鱼48h,取肝脏,运用2-DE技术及PDQuest 8.0.1软件分离分析,筛选出处理组有10个蛋白表达上调。这为NiC毒鱼机制及新型灭螺药物可能作用的特定靶蛋白提供一定的依据。
[Abstract]:Oncomelania hupensis (Oncomelania hupensis) is the only intermediate host of Schistosoma japonicum (Schistosoma japonicum). The technical and economic difficulties such as poor toxicity and high price of target / point selection. Therefore, it is necessary to develop new snail control drugs closely in combination with the national major technical requirements for schistosomiasis control. The effects of aniline NiC and water soluble steroidal saponins / alkaloids of snail killing plants on protein expression and enzyme activity of Oncomelania hupensis and Oncomelania hupensis were studied in this paper. This provides a basis for the development of novel snail-killing drugs and the selection of specific targets. 1. The optimum conditions of GOT enzyme catalytic activity of Oncomelania molluscum and liver of Oncomelania hupensis were studied by L25 (56) experiment and pole / variance analysis. The results showed that the optimum conditions of GOT catalytic activity in the liver homogenate of Oncomelania hupensis were 0.2 渭 g / L and 5.0 / 5.0 渭 mol / L, respectively, and the reaction system pH7.5/8.0, was obtained at the concentration of 0.2 渭 mol / L and 5.0 渭 mol / L, respectively. When the reaction time 60/20min and reaction temperature were 45 鈩，
本文编号：2431872
[Abstract]:Oncomelania hupensis (Oncomelania hupensis) is the only intermediate host of Schistosoma japonicum (Schistosoma japonicum). The technical and economic difficulties such as poor toxicity and high price of target / point selection. Therefore, it is necessary to develop new snail control drugs closely in combination with the national major technical requirements for schistosomiasis control. The effects of aniline NiC and water soluble steroidal saponins / alkaloids of snail killing plants on protein expression and enzyme activity of Oncomelania hupensis and Oncomelania hupensis were studied in this paper. This provides a basis for the development of novel snail-killing drugs and the selection of specific targets. 1. The optimum conditions of GOT enzyme catalytic activity of Oncomelania molluscum and liver of Oncomelania hupensis were studied by L25 (56) experiment and pole / variance analysis. The results showed that the optimum conditions of GOT catalytic activity in the liver homogenate of Oncomelania hupensis were 0.2 渭 g / L and 5.0 / 5.0 渭 mol / L, respectively, and the reaction system pH7.5/8.0, was obtained at the concentration of 0.2 渭 mol / L and 5.0 渭 mol / L, respectively. When the reaction time 60/20min and reaction temperature were 45 鈩，
本文编号：2431872
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