鼠疟原虫感染大鼠和小鼠的种特异性分析  

Specie⁃specific analysis of plasmodia infecting rats and mice

作　　者：郭帅 何彪 高源利 范永铃 朱锋 丁艳 刘太平 徐文岳 GUO Shuai;HE Biao;GAO Yuanli;FAN Yongling;ZHU Feng;DING Yan;LIU Taiping;XU Wenyue(Department of Pathogenic Biology,Army Medical University,Chongqing 400038,China)

机构地区：[1]陆军军医大学基础医学院病原生物学教研室,重庆400038

出　　处：《中国寄生虫学与寄生虫病杂志》2023年第5期539-545,551,共8页Chinese Journal of Parasitology and Parasitic Diseases

基　　金：国家自然科学基金面上项目(82172296)。

摘　　要：目的用约氏疟原虫(P.y)BY265株和伯氏疟原虫(P.b)ANKA⁃luciferase株感染小鼠和大鼠,分析鼠疟原虫感染宿主的种特异性。方法制备P.y和P.b的子孢子和裂殖子。分别用5×104的P.y和P.b子孢子经尾静脉注射感染SD大鼠、BALB/c小鼠(P.y感染组,5只/组)和SD大鼠、C57BL/6J小鼠(P.b感染组,5只/组),每日取尾静脉血涂片镜检,记录红内期疟原虫出现的时间。P.y子孢子感染SD大鼠和BALB/c小鼠后42 h取肝组织,实时荧光定量PCR(qRT⁃PCR)检测肝组织疟原虫18S rRNA的相对表达量。P.b子孢子感染SD大鼠和C57BL/6J小鼠后42 h活体成像法检测肝脏荧光值。分别用1×10^(8)、1×10^(7)、1×10^(6)的P.y和P.b被感染红细胞(iRBC)感染SD大鼠(P.y感染组、P.b感染组)和易感小鼠(BALB/c、C57BL/6J)(阳性对照组),每日取尾静脉血涂片镜检,计算原虫血症。分别用P.y、P.b裂殖子感染SD大鼠和易感小鼠,每日取尾静脉血涂片镜检,计算原虫血症;当大鼠、小鼠原虫血症达到峰值后,每隔8小时取尾静脉血涂片镜检,持续24 h,分析疟原虫发育节律;观察大鼠、小鼠实验性脑型疟(ECM)的发生情况。分别用P.y、P.b裂殖子感染T、B细胞缺陷的Rag2⁃KO SD大鼠、野生型SD大鼠和易感小鼠,每日取尾静脉血涂片观察,计算原虫血症。两组数据之间比较采用非配对t检验,组间原虫血症趋势比较采用双因素方差分析(two⁃way ANOVA)。结果P.y感染组的SD大鼠、BALB/c小鼠,P.b感染组的SD大鼠、C57BL/6J小鼠体内疟原虫均能完成肝期发育,于第3天进入红内期。qRT⁃PCR结果显示,P.y子孢子感染SD大鼠、BALB/c小鼠体内疟原虫特异性18S rRNA相对表达量分别为(1.63±0.381)、(1.00±0.232),二者差异有统计学意义(t=2.801,P<0.05)。活体成像结果显示,P.b子孢子感染SD大鼠体内荧光度值为(6.243±1.425)×10^(7),高于C57BL/6J小鼠的(1.624±0.530)×10^(7)(t=6.077,P<0.01)。红内期iRBC感染结果显示,3种剂量的P.y感染组SDObjective To analyze the infection specie⁃specificity of Plasmodium yoelii(P.y)BY265 strain and P.berghei(P.b)ANKA⁃luciferase strain in rats and mice.Methods The sporozoites and merozoites of P.y and P.b were prepared.Infection was performed by injecting 1×10^(5)P.y and P.b sporozoites to SD rats and BALB/c mice(P.y infection,5 SD rats/group,5 BALB/c mice/group)and SD rats,C57BL/6J mice(P.b infection,5 SD rats/group,5 C57BL/6J mice/group)through tail vein,respectively.The tail vein blood samples were collected daily to prepare blood mears for microscopic examination to record the appearance time of erythrosytic stage of Plasmodium.Liver tissue samples were collected at 42 h post⁃infection from the SD rats and BALB/c mice of P.y infection group to examine the relative expression level of Plasmodium 18S rRNA by real⁃time quantitative PCR(qRT⁃PCR),and measure the liver fluorescence signal in SD rats and C57BL/6J mice in P.b infection groups using IVIS Lumina imaging system.A dose of 1×10^(8),1×10^(7),1×10^(6)infected red blood cells(iRBCs)of P.y and P.b was used to infect SD rats(P.y in⁃fection group and P.b infection group)and susceptible mice(BALB/c and C57BL/6J)(positive control group),the tail vein blood was collected daily for smear to exam the parasitemia.The SD rats and susceptible mice were infected with P.y and P.b merozoites,respectively.The tail vein blood was collected daily for microscopic examination to calcu⁃late the parasitemia.When the parasitemia in rats and mice reached its peak,the tail vein blood smears were pre⁃pared every 8 hours for 24 hours for microscopic examination,and the proportions of rings,trophozoites,and schizonts were calculated to analyze the developmental rhythm of the rodent Plasmodium.The experimental cerebral malaria(ECM)in rats and mice were recorded.Rag2⁃KO SD rats,wild⁃type SD rats,and susceptible mice were infected with P.y and P.b merozoites respectively,the tail vein blood smears were examined daily to calculate parasitemia.The com⁃parison betwe

关 键 词：约氏疟原虫 伯氏疟原虫 肝期 红内期 感染 

分 类 号：R382.314[医药卫生—医学寄生虫学]