血吸虫对吡喹酮抗药性的研究ⅩⅢ日本血吸虫吡喹酮抗药性的实验诱导  

Studies on resistance of Schistosoma to praziquantel XIII Resistance of Schistosoma japonicum to praziquantel is experimentally induced in laboratory

作　　者：梁幼生[1] 李洪军[1] 戴建荣[1] 汪伟[1] 曲国立[1] 陶永辉[1] 邢云天[1] 李幼子[1] 钱科[1] 魏剑英[1] 

机构地区：[1]江苏省血吸虫病防治研究所卫生部寄生虫病预防和控制技术重点实验室江苏省寄生虫分子生物学重点实验室,无锡214064

出　　处：《中国血吸虫病防治杂志》2011年第6期605-610,共6页Chinese Journal of Schistosomiasis Control

基　　金：国家科技支撑计划(2009BAI78B06);国家自然科学基金(30471516);江苏省自然科学基金(BK2009076);江苏省卫生厅项目(X200912);江苏省预防医学研究课题(Y201031)

摘　　要：目的实验研究在吡喹酮药物压力下中国大陆日本血吸虫对吡喹酮产生抗药性的可能性。方法取采自湖南省血吸虫病流行区湖滩现场和江苏省实验室传代的感染性钉螺实验室逸蚴,获得日本血吸虫尾蚴感染小鼠,从感染鼠肝脏分离成熟虫卵孵化毛蚴感染湖北钉螺,建立现场采集株和实验室传代株日本血吸虫小鼠钉螺实验室生活史循环。用定量尾蚴（40条/鼠）感染小鼠,感染35d后将小鼠随机分为对照组和抗性诱导组：对照组小鼠感染后45d解剖收集肠系膜静脉和肝门脉静活虫体,计算虫负荷（条/鼠）;抗性诱导组小鼠采用灌胃法一次口服亚治疗剂量吡喹酮进行治疗,服药22d后解剖收集肠系膜静脉和肝门脉静存活虫体。计算虫负荷（条/鼠）和减虫率,完成首轮诱导。取抗性诱导组小鼠肝脏,分离虫卵,实验室孵化出毛蚴重新感染钉螺,感染后的钉螺经25℃生化培养箱内饲养60~70d后,分离感染性钉螺并逸蚴,用成熟尾蚴感染小鼠,开始新一轮循环诱导。首轮诱导吡喹酮口服剂量为100mg/kg,后每循环2~3轮增加100mg/kg口服剂量。取完成8轮诱导后和未经诱导原代虫株的尾蚴感染小鼠,感染35d后分别采用300mg/kg和600mg/kg吡喹酮一次性灌胃治疗感染小鼠,服药后14d解剖感染鼠,收集活虫,计算各虫株减虫率,评价虫株经8轮诱导后对吡喹酮敏感性的变化。结果在实验室内建立了江苏实验室传代株和湖南现场采集株2个虫株,并对其实施了8轮诱导。江苏实验室传代株在小鼠体内经第1轮口服100mg/kg吡喹酮治疗后减虫率为22.3%,第8轮口服300mg/kg吡喹酮治疗后减虫率为53.7%,减虫率随口服吡喹酮剂量增加而增加;湖南现场采集株在小鼠体内经第1轮口服100mg/kg吡喹酮治疗后减虫率为66.8%,第8轮口服300mg/kg吡喹酮治疗后减虫率仅为20.6%,减虫率随口服吡喹酮剂量增加而显著降低。未经吡喹酮诱导的江苏实Objective To investigate the possibility of the emergence of praziquantel resistance in Schistosoma japonicum in China's Mainland under drug pressure. Methods S. japonicum cercaria were released from the infected Oncomelania hupensis snails collected from the marshland in Hunan Province that was endemic for schistosomiasis japonica and raised in the laboratory of Jiangsu Institute of Parasitic Diseases, and mice were infected. O. hupensis snails were infected with miracidia hatched from the schistosome mature eggs that were isolated from the liver of the infected mice. The life cycles of a field isolate and a laboratory passage isolate of S. japonicum were established in laboratory via the cycle of mouse-snail. The mice were infected with 40 cercariae each, 35 days later post-infection, were grouped randomly into control and resistance-induced groups. All the mice in the control group were sacrificed on day 45 post-infection, and any adult S. japonicum worms in the hepatic and portomesenteric veins were recovered and counted, and the worm burdens were calculated. The mice in the resistance-induced group were administered orally with the sub-curative dose of praziquantel, and were sacrificed 22 days post-treatment. Any adult S. japonicum worms in the hepatic and portomesenteric veins were recovered and counted, and the worm burdens and reduction in the worms recovered which were obviously caused by the praziquantel treatment were calculated. The eggs in the liver of the mice in the resistance-induced group were isolated and hatched to yield miracidia, and then the snails were again infected with the newly hatched miracidia to complete the first-passage inducement. After raising in laboratory at 25 ℃ for 60-70 days post-infection, the infected snails were isolated and shed cercaria, and the mice were infected with the newly released cercaria to start a new passage of resistance-inducement. The oral dose of praziquantel for the first-passage inducement was 100 mg/kg, and an additional 100 mg/kg was given every 2-3

关 键 词：日本血吸虫 吡喹酮 抗药性 抗药性诱导 

分 类 号：R383.24[医药卫生—医学寄生虫学]