中华围产医学杂志
中華圍產醫學雜誌
중화위산의학잡지
CHINESE JOURNAL OF PERINATAL MEDICINE
2015年
1期
30-34
,共5页
吴德%潘娜娜%王莉莉%唐瑾%姜俊红%赵青松%李浩%卜笑松%唐久来
吳德%潘娜娜%王莉莉%唐瑾%薑俊紅%趙青鬆%李浩%蔔笑鬆%唐久來
오덕%반나나%왕리리%당근%강준홍%조청송%리호%복소송%당구래
妊娠并发症,感染性%巨细胞病毒感染%海马%学习%记忆%疾病模型,动物
妊娠併髮癥,感染性%巨細胞病毒感染%海馬%學習%記憶%疾病模型,動物
임신병발증,감염성%거세포병독감염%해마%학습%기억%질병모형,동물
Pregnancy complications,infectious%Cytomegalovirus infections%Hippocampus%Learning%Memory%Disease models,animal
目的 探讨先天性感染人巨细胞病毒(human cytomegalovirus,HCMV)的子代大鼠空间学习记忆情况及其影响机制. 方法 40只12周龄无特定病原体级Sprague-Dawley大鼠按雌雄3∶1合笼.用随机数字表法将受精大鼠分为研究组和对照组,每组15只.研究组妊娠3d腹腔接种组织半数感染量为1×10-6的病毒悬液0.5 ml;对照组同时腹腔注射人胚肺成纤维细胞上清液0.5 ml.接种后l周检测2组孕鼠外周血HCMV特异性抗体IgM和IgG.于临产前,取研究组和对照组孕鼠各3只行剖宫产,用随机数字表法在2组剖宫产所获仔鼠中各取5只,取双侧海马组织进行病毒分离.仔鼠饲养至28日龄时,从各组随机选择6只进行神经行为学检测(包括Morris水迷宫学习后的定位航行试验和空间探索试验).取完成神经行为学检测的大鼠脑组织,HE染色切片,光镜下观察;取仔鼠海马组织块透射电镜下观察、摄片.Morris水迷宫空间学习记忆测试数据采用广义混合线性模型,并用Origin 8.0绘制结果图.采用秩和检验、独立样本t检验和x2检验进行统计学分析. 结果 (1)研究组15只雌鼠中,11只成功分娩;对照组15只均成功分娩.研究组母鼠的产仔量低于对照组[(5.5±2.4)只与(8.7±3.1)只,t=2.366,P=0.033),仔鼠生后1周内死亡率高于对照组[26.1％(18/69)与4.9％(6/122),x2=191.020,P=0.000].研究组7只次海马病毒分离阳性,对照组均为阴性.(2)研究组第1～4天逃避潜伏期均长于对照组,差异有统计学意义(广义混合线性模型检验,F=499.473,P=0.000).研究组第1～4天的总路程长于对照组(广义混合线性模型检验,F=440.167,P=0.000).空间探索能力方面,研究组穿越平台次数少于对照组[(4.2l±1.44)次与(7.50±1.72)次,t=7.182,P=0.003],目标象限游泳时间占总时间比例少于对照组[(26±4)％与(47±5)％,t=15.487,P=0.000].(3)光镜观察可见研究组大鼠脑组织存在不同程度皮层液化坏死,空泡变性较多,海马结构紊乱;电镜下可见研究组大鼠脑组织存在典型的陈旧性髓鞘损伤及崩解. 结论 HCMV先天性感染仔鼠出现较明显的空间学习记忆功能下降,其病理机制主要与皮层和海马结构损伤有关.
目的 探討先天性感染人巨細胞病毒(human cytomegalovirus,HCMV)的子代大鼠空間學習記憶情況及其影響機製. 方法 40隻12週齡無特定病原體級Sprague-Dawley大鼠按雌雄3∶1閤籠.用隨機數字錶法將受精大鼠分為研究組和對照組,每組15隻.研究組妊娠3d腹腔接種組織半數感染量為1×10-6的病毒懸液0.5 ml;對照組同時腹腔註射人胚肺成纖維細胞上清液0.5 ml.接種後l週檢測2組孕鼠外週血HCMV特異性抗體IgM和IgG.于臨產前,取研究組和對照組孕鼠各3隻行剖宮產,用隨機數字錶法在2組剖宮產所穫仔鼠中各取5隻,取雙側海馬組織進行病毒分離.仔鼠飼養至28日齡時,從各組隨機選擇6隻進行神經行為學檢測(包括Morris水迷宮學習後的定位航行試驗和空間探索試驗).取完成神經行為學檢測的大鼠腦組織,HE染色切片,光鏡下觀察;取仔鼠海馬組織塊透射電鏡下觀察、攝片.Morris水迷宮空間學習記憶測試數據採用廣義混閤線性模型,併用Origin 8.0繪製結果圖.採用秩和檢驗、獨立樣本t檢驗和x2檢驗進行統計學分析. 結果 (1)研究組15隻雌鼠中,11隻成功分娩;對照組15隻均成功分娩.研究組母鼠的產仔量低于對照組[(5.5±2.4)隻與(8.7±3.1)隻,t=2.366,P=0.033),仔鼠生後1週內死亡率高于對照組[26.1％(18/69)與4.9％(6/122),x2=191.020,P=0.000].研究組7隻次海馬病毒分離暘性,對照組均為陰性.(2)研究組第1～4天逃避潛伏期均長于對照組,差異有統計學意義(廣義混閤線性模型檢驗,F=499.473,P=0.000).研究組第1～4天的總路程長于對照組(廣義混閤線性模型檢驗,F=440.167,P=0.000).空間探索能力方麵,研究組穿越平檯次數少于對照組[(4.2l±1.44)次與(7.50±1.72)次,t=7.182,P=0.003],目標象限遊泳時間佔總時間比例少于對照組[(26±4)％與(47±5)％,t=15.487,P=0.000].(3)光鏡觀察可見研究組大鼠腦組織存在不同程度皮層液化壞死,空泡變性較多,海馬結構紊亂;電鏡下可見研究組大鼠腦組織存在典型的陳舊性髓鞘損傷及崩解. 結論 HCMV先天性感染仔鼠齣現較明顯的空間學習記憶功能下降,其病理機製主要與皮層和海馬結構損傷有關.
목적 탐토선천성감염인거세포병독(human cytomegalovirus,HCMV)적자대대서공간학습기억정황급기영향궤제. 방법 40지12주령무특정병원체급Sprague-Dawley대서안자웅3∶1합롱.용수궤수자표법장수정대서분위연구조화대조조,매조15지.연구조임신3d복강접충조직반수감염량위1×10-6적병독현액0.5 ml;대조조동시복강주사인배폐성섬유세포상청액0.5 ml.접충후l주검측2조잉서외주혈HCMV특이성항체IgM화IgG.우임산전,취연구조화대조조잉서각3지행부궁산,용수궤수자표법재2조부궁산소획자서중각취5지,취쌍측해마조직진행병독분리.자서사양지28일령시,종각조수궤선택6지진행신경행위학검측(포괄Morris수미궁학습후적정위항행시험화공간탐색시험).취완성신경행위학검측적대서뇌조직,HE염색절편,광경하관찰;취자서해마조직괴투사전경하관찰、섭편.Morris수미궁공간학습기억측시수거채용엄의혼합선성모형,병용Origin 8.0회제결과도.채용질화검험、독립양본t검험화x2검험진행통계학분석. 결과 (1)연구조15지자서중,11지성공분면;대조조15지균성공분면.연구조모서적산자량저우대조조[(5.5±2.4)지여(8.7±3.1)지,t=2.366,P=0.033),자서생후1주내사망솔고우대조조[26.1％(18/69)여4.9％(6/122),x2=191.020,P=0.000].연구조7지차해마병독분리양성,대조조균위음성.(2)연구조제1～4천도피잠복기균장우대조조,차이유통계학의의(엄의혼합선성모형검험,F=499.473,P=0.000).연구조제1～4천적총로정장우대조조(엄의혼합선성모형검험,F=440.167,P=0.000).공간탐색능력방면,연구조천월평태차수소우대조조[(4.2l±1.44)차여(7.50±1.72)차,t=7.182,P=0.003],목표상한유영시간점총시간비례소우대조조[(26±4)％여(47±5)％,t=15.487,P=0.000].(3)광경관찰가견연구조대서뇌조직존재불동정도피층액화배사,공포변성교다,해마결구문란;전경하가견연구조대서뇌조직존재전형적진구성수초손상급붕해. 결론 HCMV선천성감염자서출현교명현적공간학습기억공능하강,기병리궤제주요여피층화해마결구손상유관.
Objective To investigate the influence of congenital human cytomegalovirus (HCMV) infection on spatial learning and memory skill in rats and its mechanism.Methods Forty specific pathogen free Sprague-Dawley rats (12 weeks old,male and female mated by 1:3) were selected and randomly divided into the experimental group and control group,fifteen in each group.Fertilized female rats in the experimental group were injected with 0.5 ml HCMV (1 × 10-6 of the 50％ tissue culture infective dose/rat) by intraperitoneal inoculation on the third gestational day.At the same time,a 0.5 ml suspension of human embryonic lung fibroblast cells was inoculated intraperitoneally into fertilized female rats in the control group.HCMV-specific IgM and IgG in maternal peripheral blood of the two groups were detected on one week after inoculation.Both side of hippocampus of ten neonatal rats (five in each group),born by cesarean section,were collected for virus isolation.Spatial learning and memory skill in 4-6 week offspring in the two groups were assessed using the Morris Water Maze (MWM) test,and results were presented by Origin 8.0.Pathological damage in offspring was determined by microscopy and electron microscopy.Independent samples t-test,Chi square and rank sum tests were used for statistical analysis.Results (1) Compared with the control group,litter size and mortality of neonatal rats within one week after birth in the experimental group were significantly higher [litter size:(8.7± 3.1) vs (5.5±2.4),t=2.366,P=0.033; mortality of neonatal rats:4.9％ (6/122) vs 26.1％ (18/69),x2=191.020,P=0.000.In the MWM test,escape latency in 30-day old offspring in the experimental group was significantly increased compared with the control group during 4 days of training (F=499.473,P=0.000).The total distance in the experimental group was significantly increased compared with the control group (F=440.167,P=0.000).In addition,the numbers of platform crossing (4.21 ± 1.44) and swimming time in the target quadrant [(26±4)％] in the congenital infection group were decreased compared with the control group [(7.50± 1.72) and (47 ± 5)％,t=-7.182 and 15.487,P=0.003 and 0.000].In the congenital infection group,liquefaction necrosis of the cerebral cortex (light microscopy) and myelin disintegration in the hippocampus (electron microscopy) were observed.Conclusions Injury of the cortex and hippocampus in offspring with congenital HCMV infection is involved in the pathological mechanism of decreased spatial learning and memory skill.
