腹泻儿童鼠伤寒沙门菌spv对自噬的作用及其分子机制的研究

项目名称： 腹泻儿童鼠伤寒沙门菌spv对自噬的作用及其分子机制的研究

项目编号： No.81460117

项目类型： 地区科学基金项目

立项/批准年度： 2015

项目学科： 医药、卫生

项目作者： 陈强

作者单位： 江西省儿童医院

项目金额： 47万元

中文摘要： 沙门菌感染所致疾病严重危害儿童健康，适度的自噬有利于机体对沙门菌感染的防御，多种信号通路在其中发挥重要作用。沙门菌的毒力基因spv能调控p38、NADPH等自噬的关键因子。那么，spv能否影响自噬？前期研究发现，从江西地区腹泻儿童中分离的沙门菌中，鼠伤寒沙门菌是主要的血清型，利用自杀质粒构建的spvB缺陷菌株中自噬体标记蛋白LC3Ⅱ及自噬相关基因Beclin-1表达上调，提示spv表达的改变可以调节宿主细胞的自噬。为此，本课题拟以江西南昌地区腹泻儿童临床样本中分离的spv高表达的鼠伤寒沙门菌为研究对象，通过自杀质粒构建基因缺陷变异株，利用电转化制备缺陷变异株回补株，体内外实验观察下调spv表达后对自噬、自噬相关基因、信号通路的影响及信号通路间相互作用，并探讨可能机制，为沙门菌感染的临床治疗提供新的研究思路和靶点，为进一步探讨耐药及非耐药沙门菌株中spv对自噬影响的差异及其分子机制奠定基础。

中文关键词： 腹泻儿童；分子机制；自噬；spv；鼠伤寒沙门菌

英文摘要： Salmonellosis does great harms to children's health.Moderate autophagy is helpful to the host defense to the infection caused by salmonella，in which various signaling pathways play a pivotal role.Spv virulence gene can regulating the autophagy-associated factors, including p38 and NADPH.However,could spv exert a role in autophagy? Previous studies suggested that S. Typhirium is the main serotype in salmonella isolated from the children with diarrhea in Jiangxi district. In suicide plasmid constructed spvB-deficient mutant strain,the expression of autophagosome-specific marker protein LC3Ⅱ and autophagy-related gene Beclin-1 was up-regulated,which indicated that the change of spv expression could regulate autophagy.In this research, we plan to utilize human gastrointestinal epithelial cells and S. Typhirium we isolated before, construct spvBC-deficient mutant strain using suicide plasmid, establish spvBC retro-complementary strain by electroporation and carry out in vivo and vitro trials in order to clarify the influence of spv in autophagy , autophagy-related genes and signaling pathways,as well as the interaction of these pathways,to discuss the potential mechanism for the purpose of providing new research approach and target for guiding the clinical treatment to Salmonellosis and setting up the molecular basis for Subsequent research on the difference of the influence of spv in autophagy between antibiotic-resistant S. Typhirium and non-resistant S. Typhirium and the underlying mechanism.

英文关键词： diarrheic children;molecular mechanism;autophagy;spv;S.Typhirium