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目的了解半胱氨酸蛋白酶(CP)基因在猬迭宫绦虫成虫及裂头蚴不同部位及裂头蚴不同感染时期的时空表达模式。方法收集安顺地区野生王锦蛇体内猬迭宫绦虫裂头蚴。将24只昆明小鼠随机分成8组,每组3只,以5条裂头蚴/只感染小鼠,于感染后30min~14d剖杀1组小鼠,收集感染小鼠体内的裂头蚴,以10条/只感染3只幼犬,于感染后1个月剖杀幼犬,收集成虫。利用实时定量PCR检测猬迭宫绦虫成虫和裂头蚴不同部位及不同感染时期裂头蚴CP基因的表达情况。结果小鼠经口感染裂头蚴30min后腹腔内查见穿过胃肠壁的裂头蚴;感染后30min~6h,裂头蚴主要见于胃肠腔、胃肠壁和腹腔内;感染24h,有少量的裂头蚴移行至小鼠皮下肌肉组织;感染7d后,多数裂头蚴移行至皮下肌肉组织寄生。实时荧光定量PCR检测CP基因在成虫及裂头蚴的不同部位均有表达,但成虫不同部位的表达量均低于幼虫时期。成虫不同部位的表达量依次为孕节>头节>成节,裂头蚴不同部位的表达量依次为体段>头段>尾段。裂头蚴感染小鼠30min时CP基因表达量下调,感染6h时达到峰值,随后下降,感染14d时降至最低。结论 CP基因在猬迭宫绦虫裂头蚴感染初期及裂头蚴体段和成虫孕节高表达,提示CP可能参与猬迭宫绦虫的生长、发育、入侵及移行等过程。
Objective To investigate the temporal and spatial expression pattern of cysteine protease (CP) gene in different stages of adult Tissue and adult sporozoite. Methods The wild Eupolyphaga henrymans were collected from the wild Wang Jin snake in Anshun area. Twenty-four Kunming mice were randomly divided into 8 groups with 3 mice in each group. Five mice were challenged with 5 splenicclasts per mouse, and one group of mice were killed 30 to 14 days after infection. Spleniccosis was collected in 10 mice / Infected only 3 puppies, 1 month after infection, puppies, collect adult. Real-time quantitative PCR was used to detect the expression of CP gene in different parts of adult and sporozoite larvae of the hedgehog and different stages of infection. Results The mice were infected orally by spleen larvae for 30min and the splenic cercariae penetrating the gastrointestinal wall were observed intraperitoneally. After 30min ~ 6h infection, the spleniccrocytes were mainly found in the gastrointestinal tract, gastrointestinal wall and abdominal cavity. A small amount of fission larvae migrated to Mouse subcutaneous muscle tissue; 7d after infection, most of the migrating spleen cell subcutaneous muscle tissue parasitization. Real-time PCR showed that CP gene was expressed in different parts of adult and sporozoite, but the expression of CP in different parts of adult was lower than that in larval stage. The expression levels of different parts of adult worms were in order of pregnancy node> head node> nodule, and the expression of different parts of sporozoite in order of body segment> head segment> tail segment. The expression of CP gene was down-regulated in mice infected with sporozoite at 30min, peaked at 6h, then decreased and then decreased to the lowest at 14d. CONCLUSION: The CP gene is highly expressed in the early stage of infection and in the second stage of adult metacercaria and second stage adult female of metacercaria of Toxoplasma gondii, suggesting that CP may be involved in the growth, development, invasion and migration of Toxoplasma gondii.