【作者】 赵云鹤；

【导师】 侯玉英；

【作者基本信息】 山西医科大学 ， 病原生物学， 2006， 硕士

【摘要】 目的:本实验采用弓形虫感染BALB/c雌鼠,①观察染虫雌鼠的毒性反应,建立生殖毒性的小鼠模型;②染虫雌鼠排卵情况及卵母细胞染色体的变化;③染虫雌鼠性激素(FSH、LH、E2)水平的变化;④着床前受精卵的发育情况;⑤孕中期鼠染虫后胚胎的发育情况,探讨弓形虫对雌鼠生殖功能及胚胎发育的毒性作用。方法:(1)建立生殖毒性的小鼠模型:将BALB/c雌鼠(21±0.5g,9-10周)随机分为11组,每组10只:A组正常对照组(PBS)、染虫B组(腹腔注射经胰酶消化后的弓形虫速殖子25个/只/2组、50个/只/3组、100个/只/4组、200个/只/5组、400个/只/6组)和染虫C组(腹腔注射不经胰酶消化的弓形虫速殖子,设剂量、组数相同)。观察小鼠生存情况(20天)。(2)弓形虫对卵母细胞的影响:将BALB/c处女鼠随机分为:正常对照1组(PBS),染虫组(腹腔注射纯化的弓形虫速殖子:25个/只/2组、50个/只/3组、100个/只/4组、200个/只/5组),每组10只;染虫3天后超排,取卵母细胞计数、观察其染色体。(3)弓形虫对雌鼠性激素分泌的影响:设正常对照组(PBS)和染虫组(腹腔注射纯化的弓形虫速殖子200个/只),染虫6天后,摘眼球取血,分离血清,用放免法测定雌鼠在一个动情周期中FSH、LH的水平;并测定雌鼠妊娠前3天E2的水平。(4)弓形虫对受精卵发育的影响:将BALB/c鼠随机分为正常对照组(PBS)和染虫组(腹腔注射纯化的弓形虫速殖子:25个/只/2组、50个/只/3组、100个/只/4组、200个/只/5组),3天后超排、合笼、交配,次晨查阴栓,取各组受精鼠,再随机分3组,每组10只。观察妊娠前3天的受精卵、2-细胞胚、4-细胞胚、桑椹胚及未受精卵、1-细胞胚、延迟胚,退化胚的数量、比率,并取小鼠主要脏器作病理切片,观察其病理学变化。(5)弓形虫感染孕中期BALB/c鼠的胚胎毒性:将受精鼠随机分为阴性对照组(PBS,1组)、染虫组(2～5组)和环磷酰胺组(6组),每组10只,孕8天染虫(25个/只/2组、50个/只/3组、100个/只/4组、200个/只/5组),孕18天颈椎脱臼处死孕鼠,称雌鼠子宫、胎盘、胎鼠重量。观察胎鼠有无吸收胎、死胎,并计数,取出活胎,肉眼观察其外观、内脏、骨骼有无畸形。结果:(1)胰酶消化组的生存天数显著高于未消化组(P<0.01),且胰酶消化各组间均有差异(P<0.05);雌鼠染虫25个/只无毒性反应;染虫400个/只,毒性较大,实验中全部死亡,可视为致死量。且随着剂量的增加,生存时间缩短。(2)染虫组:25个/只/2组、50个/只/3组、100个/只/4组、200个/只/5组的卵母细胞回收数均低于正常对照1组(P<0.05),3～4组染色体畸变率均高于正常组,但无显著性差异更多还原

【Abstract】 Objective:This study was performed to research the reproductive toxicity on BALB/c female mice infected with toxoplasma gondii.①To observ the toxicity of infected mice and establish mice reproductive toxicity model;②To observe the infected BALB/c female mice ovulation quantums and ovum chromosome;③To observe the oestrogenic hormone( FSH, LH, E2 )level of infected BALB/c female mice;④To observe zygote developmental condition before nidation;⑤To observe embryonic development condition of infected mid-gravidity mice to research the reproductive toxicity and development toxicity of toxoplasma gondii to the BALB/c female mice.Methods:This study included five parts:The first part: To create the reproductive toxicity mice model: The healthy BALB/c mice were divided into 11 groups randomly which every group was 10: the normal control group injected with PBS (A), the infected group B (trypsinization group, 25, 50, 100, 200, 400 Toxoplasma gondii tachyzoites every group); the infected group C of giving the same doses as B (no trypsinization). Then we observed the mice survival state (20 days) and record foodintake, waterintake everyday and piloerection, idle motion, shiver, fastidium, death.The second part: To observe the influence of toxoplasma gondii on ovum of female mice infected. Take 50 health virgin mice, divided into 5 groups in random: the normal control group and the infected groups (25, 50, 100, 200 toxoplasma gondii tachyzoites). 3 days after infection, take ovum in anatomical lens and observe, count. Make ovum chromosome, to observe chromosome strip number and leakage, and so on.The third part: To observe the influences by toxoplasmosis to female hormone: set up the normal control group (PBS) and the toxoplasma infected group (200 tachyzoites), after 6 days, take the blood, separate the serum, determine the FSH, LH level in one estrous cycle; evaluate the E2 in the pregnant first 3 days.The fourth part: This study was performed to study the toxicity of toxoplasma gondii infection to the fertilized ovum: Divided the BALB/c virgin mice into 5 groups, including the infected group (25, 50, 100, 200 tachyzoites) and the normal control group in random. Mice infected was gived homologous dose toxoplasma gondii tachyzoites by tne intraperitoneal injection, each mice intraperitoneal injected 0.2ml, while mice peritoneal injection administrated with PBS were control. The day inoculated was recorded with 0 day. The mice were更多还原