豚鼠胎仔肾脏巨细胞病毒感染的实验研究

The experimental study of guinea pig cytomegalovirus infection in the kidney of the pup of guinea pig

目的 探讨豚鼠胎仔肾脏巨细胞病毒感染与妊娠结局的关系。方法 随机选择孕早期豚鼠 2 0只 ,腹腔接种巨细胞病毒 ,观察其妊娠结局 ,分娩后 2 4h内处死新生豚鼠。用地高辛标记的巨细胞病毒晚期mRNA三相探针进行原位杂交 ,检测豚鼠胎仔肾脏内巨细胞病毒负荷量及分布情况。结果 2 0只母豚鼠共妊娠 6 3只胎仔 ,其中足月活产并存活到 2 4h以上的胎仔 4 2只 ,流产等异常胎仔 2 1只。原位杂交检测正常胎仔巨细胞病毒感染率 7 1% (3/ 4 2 )、平均吸光度 (A)为 0 10 5±0 0 5 2 ,异常胎仔巨细胞病毒感染率 2 8 6 % (6 / 2 1)、平均A为 0 15 8± 0 0 4 7,两者平均A比较 ,差异有显著性 (P <0 0 5 )。杂交阳性信号主要分布在肾小管和集合管上皮细胞。结论 巨细胞病毒晚期mRNA主要在肾小管和集合管上皮细胞中表达 ,其表达水平与异常妊娠结局相关。

Objective To study the relationship of guinea pig cytomegalovirus (GPCMV) infection with the outcome of pregnancy by the kidney of guinea pig(GP). Methods Twenty first-trimester gestation GPs were randomly selected, intraperitoneally inoculated with GPCMV. Then female GPs and the pups were killed within 24 h after delivery. By in situ hybridization(ISH) with three phases GPCMV late-mRNA probes labeled by digoxin, the virus load and its distribution were screened inside the pup′s kidney. Results Twenty GPs totally conceived 63 pups. Among them, 42 had normal outcome and lived longer than 24 h; 21 had abnormal outcome such as abortion, fetal death, et al. By in situ hybridization, the infection rate of normal pups was 7.1%(3/42) and the average optical density( A ) was 0.105±0.052. The infection rate of abnormal pups was 28. 6%(6/21) and the A was 0.158±0.047. The difference of the A was significant ( t =2.57, P <0.05). The positive signal of ISH was mainly distributed in the epithelium of renal tubule and collecting duct. Conclusion It is concluded that the late-mRNA mainly expressed in the epithelium of renal tubule and collecting duct and the expression level was related with the abnormal pregnancy outcome.