大肠埃希菌对细粒棘球蚴活性影响的体外研究  

In vitro study of the pathway associated with the death of Echinococcus granulosus by Escherichia coli

作　　者：苗章 程萧 满洋 林芷伊[2] 吴向未[2] 伍希雅 蔡克丰 崔杰 张宏伟[2,3] MIAO Zhang;CHENG Xiao;MAN Yang;LIN Zhiyi;WU Xiangwei;WU Xiya;CAI Kefeng;CUI Jie;ZHANG Hongwei(School of Medicine,Shihezi University,Shihezi,Xinjiang 832008,China;First Affiliated Hospital,School of Medicine,Shihezi University,Shihezi,Xinjiang 832008,China;NHC Key Laboratory of Prevention and Treatment of Central Asia High Incidence Disease,Shihezi,Xinjiang 832008,China)

机构地区：[1]石河子大学医学院,新疆石河子832002 [2]石河子大学医学院第一附属医院,新疆石河子832008 [3]国家卫生健康委中亚高发病防治重点实验室,新疆石河子832000

出　　处：《石河子大学学报（自然科学版）》2023年第1期100-105,共6页Journal of Shihezi University(Natural Science)

基　　金：国家自然科学基金项目(81860363);中国医学科学院中央级公益性科研院所基本科研业务费专项资金(2020-PT330-003)。

摘　　要：目的研究细粒棘球蚴与特定浓度大肠埃希菌体外共培养的条件下活性、形态随时间的变化,以及部分参与通路。方法将羊源性原头蚴重悬计数,分为空白组、0.5麦氏浓度大肠埃希菌组(实验组),并以12、24、48、72、96 h为时间节点,使用0.1%伊红染液按照体积1∶1的比例染色2 min后显微镜下观察并计算两组原头蚴的存活率;使用活性氧试剂盒在上述时间点检测两组原头蚴体内ROS水平;在12、48、96 h分别检测Caspase-3、9;Bax、Bcl-2、Nrf2蛋白的表达水平。结果实验组原头蚴存活率随时间延长降低,在96 h降至为0%;两组原头蚴体内活性氧检测显示:实验组ROS水平明显提高,12 h最高,差异具有统计学意义(T=11.638,P<0.05),在72 h ROS水平与空白组相近;Western blot实验表明,凋亡蛋白Bax(T=4.997,P<0.05)、Caspase-3蛋白(T=6.118,P<0.05)、Caspase-9蛋白(T=3.435,P<0.05),实验组的蛋白水平大于空白组,有统计学意义。抗凋亡蛋白Bcl-2,实验组的蛋白水平小于空白组,有统计学意义(T=9.184,P<0.05),两组Nrf2蛋白水平差异无统计学意义(T=0.902,P=0.418)。结论0.5 mol·L^(-1)大肠埃希菌在体外可以快速致细粒棘球蚴死亡,氧化应激通路可能参与了此过程。Objective To study the changes in activity and morphology over time of Echinococcus granulosus co-cultured with specific concentrations of Escherichia coli in vitro,and some of the pathways involved.Methods The sheep-derived protozoa were resuspended and counted,divided into blank group and 0.5 maixa concentration Escherichia coli group(experimental group),and stained with 0.1%eosin staining solution at a ratio of 1∶1 by volume for 2 min at 12,24,48,72,96 h as time points and then observed under microscope and calculated the survival rate of protozoa in both groups;the reactive oxygen species kit was used to detect the protozoa in both groups at the above time points The expression levels of Caspase-3 and 9,Bax,Bcl-2 and Nrf2 were measured at 12,48,96 h.Results The survival rate of protozoa in the experimental group decreased with time,and decreased to 0% at 96 h.The detection of reactive oxygen species in the two groups showed that the level of ROS in the experimental group increased significantly,with the highest level at 12 h.The difference was statistically significant(T=11.638,P<0.05),and the level of ROS at 72 h was similar to that of the blank group.Western blot experiment showed that the protein levels of apoptotic protein Bax(T=4.997,P<0.05),Caspase-3 protein(T=6.118,P<0.05),Caspase-9 protein(T=3.435,P<0.05),the protein levels in the experimental group were greater than those in the blank group,which was statistically significant.For anti-apoptotic protein Bcl-2,the protein level of the experimental group was smaller than that of the blank group,which was statistically significant(T=9.184,P<0.05),and the difference in Nrf2 protein level between the two groups was not statistically significant(T=0.902,P=0.418);Conclusion Escherichia coli(0.5 mol·L^(-1))can rapidly cause death of Echinococcus granulosus in vitro,The oxidative stress pathway may be involved in this process.

关 键 词：细粒棘球蚴 大肠埃希菌 活性氧(ROS) BCL-2蛋白 Nrf2蛋白 

分 类 号：R532.32[医药卫生—内科学]