胚胎干细胞发育相关基因PLK1在细粒棘球绦虫体外不同发育时期差异表达研究  

Differential expression of stem-cell-related PLK1 gene at different developmental stages of Echinococcus granulosus in vitro

作　　者：帕力塔吉·麦麦提祖农 吴川川 田梦潇 齐文静 艾柯代·卡得 焦红杰 龚巧巧 李军 张文宝 张亚楼[1,2] Palitaji·Maimaitizunong;WU Chuan-chuan;TIAN Meng-xiao;QI Wen-jing;Aikedai·Kade;JIAO Hong-jie;GONG Qiao-qiao;LI Jun;ZHANG Wen-bao;ZHANG Ya-lou(School of Basic Medicine,Xinjiang Medical University,Urumqi 830017,China;State Key Laboratory of Pathogenesis,Prevention and Treatment of High Incidence Diseases in Central Asia,First Affiliated Hospital of Xinjiang Medical University,Urumqi 830054,China)

机构地区：[1]新疆医科大学基础医学院,乌鲁木齐830017 [2]新疆医科大学第一附属医院,临床医学研究院,中亚高发病成因与防治国家重点实验室,乌鲁木齐830054

出　　处：《中国人兽共患病学报》2023年第8期719-726,共8页Chinese Journal of Zoonoses

基　　金：国家自然科学基金(No.81830066)。

摘　　要：目的探究PLK1基因在细粒棘球绦虫不同发育时期的表达情况。方法体外培养细粒棘球绦虫原头蚴在犬胆汁刺激下向成虫方向发育,收集不同发育时期的虫体,通过免疫组化定位PLK1基因的表达。光学显微镜下切割分离成虫头节和后颈节段,收集样本运用荧光定量qPCR量化PLK1基因在虫体不同发育时期或不同部位的表达量水平,用SPSS 26.0软件进行单因素方差分析。结果成功建立了细粒棘球绦虫原头蚴向成虫方向发育的体外培养模型,体外培养的虫体56 d获得4个节片;PLK1基因在细粒棘球绦虫包囊生发层中高表达量。虽然在成虫和原头蚴表达较低,但免疫组化显示在原头蚴吸盘的底部和成虫头节的下部链体延长区域的表达高于其他部位(F=14.87,P<0.05)。结论细粒棘球绦虫干细胞相关基因PLK1的表达明显位于生发层细胞和激活原头蚴的头节底部以及链体增长区域,PLK1是棘球绦虫成虫发育干细胞区域的高表达基因。This study aimed to identify PLK1 expression in different stages and tissues of Echinococcus granulosus.To obtain adult worms in vitro,we stimulated protoscoleces(PSCs)of E.granulosus with dog bile and performed in vitro culture.The worms were cut into two parts—head-neck and body—under a light microscope.Quantitative PCR was used to determine the PLK1 gene transcription level.Immunohistochemistry was used to probe the localization of PLK1 in PSCs and adult worm tissue sections.PSCs were successfully evaginated after short stimulation with distilled water and developed to the four segment stage after 56 days of culture in vitro.PLK1 was highly expressed in the germinal cells of cysts.Although this gene had low expression in PSCs and adult worms,it was highly expressed in the bottom region of scolices.These results indicated that the PLK1 gene is likely to be an embryo-associated gene and is highly expressed in the germinal cells of both the larval cyst and adult tapeworm stages of E.granulosus.

关 键 词：细粒棘球绦虫 体外培养 PLK1 免疫组化 荧光定量PCR 

分 类 号：R383.3[医药卫生—医学寄生虫学]