The proteins from endosperm, root, stem, leaf, leafstalk of Jatropha curcas L. and their calli were hybridized to the seed toxin protein (curcin) of Jatropha curcas L. by western blot. The result showed that the curci...The proteins from endosperm, root, stem, leaf, leafstalk of Jatropha curcas L. and their calli were hybridized to the seed toxin protein (curcin) of Jatropha curcas L. by western blot. The result showed that the curcin was specifically expressed in the endosperm and its calli, while it was not detected in root, stem, leaf, and leafstalk of Jatropha curcas L. and their calli . This study indicated that calli induced from endosperm can be used to produce curcin.展开更多
文摘目的基于定量蛋白质组学探讨补肾益精方治疗2型糖尿病骨质疏松症(type 2diabetic mellitus osteoporosis,T2DOP)可能涉及的药理作用及机制。方法将60只SPF级雄性Wistar大鼠随机选取10只作空白组,其余50只大鼠参照唐辉的方法通过腹腔注射链脲佐菌素构建T2DOP大鼠模型。将符合T2DOP模型的大鼠随机分为模型组、阳性对照组、补肾益精方低、中、高剂量组,分别给予生理盐水、阿仑膦酸钠和低、中、高剂量补肾益精方灌胃干预8周。期间记录大鼠一般情况、血糖和体质量的变化;干预结束后取大鼠股骨使用Micro-CT进行扫描,并对骨松质进行3D重建和参数分析;苏木精—伊红染色观察大鼠股骨组织形态改变。根据Micro-CT及苏木精—伊红染色结果,选择补肾益精方中一组与模型组、空白组以稳定同位素二甲基化标记的方法运用定量蛋白质组学技术探究补肾益精方干预T2DOP的蛋白网络及作用机制筛选出差异蛋白,蛋白质印迹法、免疫组化检测验证差异蛋白。结果(1)Micro-CT扫描3D重建及苏木精—伊红染色结果显示,补肾益精方可以改善T2DOP大鼠模型的骨代谢平衡、改善骨微结构和增强骨力学性能。(2)通过蛋白质组学得出,空白组与模型组比较,筛选到107差异蛋白(上调蛋白82个,下调蛋白24个);补肾益精方高剂量组与模型组比较,筛选到78个差异蛋白(上调蛋白15个,下调蛋白63个)。对筛选的差异蛋白分别进行了基因本体论(Gene Ontology,GO)和京都基因与基因组百科全书(KyotoEncyclopedia of Genes and Genomes,KEGG)分析和蛋白质—蛋白质相互作用(Protein-ProteinInteraction,PPI)网络分析,发现23个差异蛋白之间存在相互作用,形成了一个复杂的调控网络,网络中的核心蛋白包括MMP9、TNF、Akt1,这些核心蛋白在PPI网络中处于枢纽位置。结论补肾益精方通过MMP9、TNF、Akt1等核心基因蛋白的表达,抑制骨吸收,促进骨形成,从而达到防治T2DOP目的。
文摘The proteins from endosperm, root, stem, leaf, leafstalk of Jatropha curcas L. and their calli were hybridized to the seed toxin protein (curcin) of Jatropha curcas L. by western blot. The result showed that the curcin was specifically expressed in the endosperm and its calli, while it was not detected in root, stem, leaf, and leafstalk of Jatropha curcas L. and their calli . This study indicated that calli induced from endosperm can be used to produce curcin.
