人巨细胞病毒孕早期先天性感染模型子代大鼠孤独症谱系障碍症状研究

Offspring symptoms of autism spectrum disorder in the rat’s SD model of HCMV congenital infection

目的:建立人巨细胞病毒(HCMV)孕早期宫内感染模型,探讨子代大鼠ASD症状。探讨HCMV孕早期先天性感染致子代孤独症谱系障碍(ASD)症状。方法:随机选取孕3天SD大鼠14只,实验组7只腹腔注射1×10^-6TCID50病毒悬液0.5ml,对照组7只腹腔注射人胚肺成纤维细胞(HELF)上清液0.5ml。仔鼠中随机各取6只,实验组母鼠的仔鼠记为HCMV组(n=6),对照组母鼠的仔鼠记为HELF组(n=6),分别进行病理及行为学试验。结果:病理及电镜结果显示实验组子代大鼠海马髓鞘破坏。Morris游泳距离:HCMV组1—4天总路程(d1:17.97±3.82;d2:15.22±2.96;d3:14.22±2.08;d4:10.87±2.34)m,高于HELF组1-4天总路程(d1:9.36±1.95;d2:6.31±1.65;d3:5.36±1.23;d4:5.01±1.24)m,差异有显著性意义(P<0.01)。刻板行为结果:HCMV组刻板动作次数(40.7±3.98,38.7±3.4,38.3±4.13,40.2±3.4,39.7±2.01,38.5±3.00)次,明显多于HELF组(18.5±2.70,21.7±3.17,26.4±3.26,20.2±2.29,19.2±2.94,20.9±3.00)次,差异有显著性意义(P<0.01)。旷场实验结果:HCMV组站立次数(19.15±3.63)次、跨格子数(41.70±11.05)均显著低于HELF组站立次数(22.05±4.14)次、跨格子数(66.05±12.57)(P<0.05)。悬吊试验HCMV组仔鼠得分(1.52±0.61)显著低于对照组(2.70±0.73)(P<0.01),斜坡试验HCMV组仔鼠评分(1.96±0.29)显著低于HELF组(4.25±0.48)(P<0.01)、倾斜板试验HCMV组停留时间(48.17±7.15)s显著低于HELF组(63.99±5.76)s(P<0.01)。结论:HCMV孕早期感染,子代大鼠有ASD样症状,表明HCMV孕早期宫内感染可能是诱发ASD的病因之一。

Objective:The intrauterine infection model of HCMV in early pregnancy was established to investigate the ASD symptoms of offspring rats.Method:Fourteen SD rats with 3 days pregnant were randomly selected:7 rats in the experimental group were intraperitoneally injected with 0.5ml of 1×10^-6TCID50 virus suspension,and 7 rats in the control group were intraperitoneally injected with 0.5 ml supernatant of human embryonic lung fibroblast(HELF).Six offspring rats were randomly selected from each group.The offspring rats of the experimental group were recorded as the HCMV group(n=6),and the offspring rats of the control group were recorded as the HELF group(n=6).Pathological and behavioral tests were conducted respectively.Result:Pathological and electron microscopy results showed that the hippocampal myelin sheath of the offspring of the experimental group was destroyed.Morris swimming distance:1—4 days total distance of HCMV group(d1:17.97±3.82;d2:15.22±2.96;d3:14.22±2.08;d4:10.87±2.34)m,was higher than that of HELF group(d1:9.36±1.95;d2:6.31±1.65;d3:5.36±1.23;d4:5.01±1.24)m,and the difference was significant(P<0.01).Stereotyped behavior results:the number of stereotyped actions in the HCMV group(40.7±3.98,38.7±3.4,38.3±4.13,40.2±3.4,39.7±2.01,38.5±3.00)times was significantly more than the HELF group(18.5±2.70,21.7±3.17,26.4±3.26,20.2±2.29,19.2±2.94,20.9±3.00)times,and the difference was significant(P<0.01).Open field experiment results:the number of standing times(19.15±3.63)and the number of cross lattices(41.70±11.05)in the HCMV group were significantly lower than the number of standing times(22.05±4.14)and the number of cross lattices(66.05±12.57)in the HELF group(P<0.05).The suspension test:The score of offspring in the HCMV group(1.52±0.61)was significantly lower than that in the control group(2.70±0.73)(P<0.01),and the score of the slope test in the HCMV group(1.96±0.29)was significantly lower than that in the HELF group(4.25±0.48)(P<0.01).In the inclined plate test,the residence time of HCMV group(48.17±7.15)s was significantly lower than HELF group(63.99±5.76)s(P<0.01).Conclusion:Early infection of HCMV pregnancy and asd-like symptoms in offspring rats suggest that early intrauterine infection of HCMV pregnancy may be one of the causes of ASD induction.