细粒棘球绦虫Innexin-unc9基因在不同发育阶段的表达分析  

Expression analysis of Innexin-unc9 gene in E.granulosus at different developmental stages

作　　者：黎飞海 吴军[1] 张文宝 杨梅 LI Feihai;WU Jun;ZHANG Wenbao;YANG Mei(Labour and Environmental Hygiene Department,College of Public Health,Xinjiang Medical University,Urumqi 830011,China;Department of Biochemistry and Molecular Biology,School of Basic Medical Sciences,Xinjiang Medical University,Urumqi 830011,China;State Key Laboratory of Pathogenesis,Prevention and Treatment of High Incidence Diseases in Central Asia,Clinical Medicine Institute of First Affiliated Hospital,Xinjiang Medical University,Urumqi 830011,China;Xinjiang Key Laboratory Biology for Endemic Diseases,Xinjiang Medical University,Urumqi 830011,China)

机构地区：[1]新疆医科大学公共卫生学院劳动卫生与环境卫生教研室,乌鲁木齐830011 [2]新疆医科大学基础医学院生物化学与分子生物学教研室,乌鲁木齐830011 [3]新疆医科大学第一附属医院临床医学研究院省部共建中亚高发病成因与防治国家重点实验室,乌鲁木齐830011 [4]新疆医科大学新疆地方病分子生物学重点实验室,乌鲁木齐830011

出　　处：《新疆医科大学学报》2022年第10期1085-1090,共6页Journal of Xinjiang Medical University

基　　金：新疆维吾尔自治区自然科学基金(2017D01C197);新疆医科大学博士科研启动金(2019-03)。

摘　　要：目的探究胆汁酸盐(胆盐)对细粒棘球绦虫(E.granulosus,E.g)发育分化的影响,分析E.gInnexin-unc9(EgInx)基因在体外有无胆盐-牛磺胆酸钠(Sodium taurocholate)培养的原头蚴(PSC)的表达水平,为研发犬抗E.g疫苗奠定基础。方法采用含与不含T4009(牛磺胆酸钠的商品型号缩写)的单相培养基分别进行PSC培养,在不同培养时间点对比分析PSC外翻率和发育状态;利用反转录荧光定量PCR(RT-qPCR)和Western Blotting检测不同培养点样本的EgInx相对表达量。结果不同时间点平行培养的原头蚴外翻情况不同,在培养30 min时和10 d以后,无T4009组的PSC外翻率急剧下降;RT-qPCR结果显示:与从体内刚采集的PSC相比,其EgInx的表达量在培养30 min的T4009组较高,且此时T4009组表达量高于平行无T4009组,且差异具有统计学意义(P<0.05),培养到17 d时,得到类似的结果;Western Blotting和RT-qPCR结果一致。结论提示在有牛磺胆酸钠的环境中,E.gInnexin-unc9在关键节点表达升高,可能参与调控细粒棘球绦虫的生长发育。Objective To explore the effect of bile salt on differentiation and development of E.granulosus,and to analyze the expression level of EgInnexin-unc9(EgInx)gene of PSC in vitro at different developmental stages,and to lay the foundation for development of canine vaccine.Methods PSC culture was carried out in monophasic medium with and without T4009(Sodium Taurocholate),and PSC ectropion rates and developmental status were compared at different culture time points.Reverse transcription fluorescence quantitative PCR(RT-qPCR)and Western Blotting were used to detect the relative expression of the EgInx expression in different culture sites.Results Parallel culture was carried out with and without T4009(Commodity model abbreviation for sodium taurocholate),and PSC was continuously cultured for 17 days.Its eversion was different at different time points.At 30 min and after 10days,the valgus rate of the PSC in the T4009-free medium was dropped sharply.The results of RT-qPCR showedthat the level ofEgInxexpression was higher at 30 min than other points of time in vitro culture,and theEgInxex-pression level of T4009 group was higher than that of parallel non-T4009 group at 30 minutes of the culture,andstatistical same results were observed at 17 days of culture(P<0.05).The results of Western Blotting were consist-ent with RT-qPCR.ConclusionThe results suggest that the elevated expression of EgInnexin unc9 at key time pointsmay be involved in regulating the differentiation and development ofE.granulosusunder the bile salt environment.

关 键 词：细粒棘球绦虫 牛磺胆酸钠 内联蛋白 表达 

分 类 号：R383.33[医药卫生—医学寄生虫学]