中华围产医学杂志
中華圍產醫學雜誌
중화위산의학잡지
CHINESE JOURNAL OF PERINATAL MEDICINE
2013年
11期
677-681
,共5页
巨细胞病毒感染%淋巴细胞%Toll样受体2%Toll样受体9%髓样分化因子88%细胞因子类
巨細胞病毒感染%淋巴細胞%Toll樣受體2%Toll樣受體9%髓樣分化因子88%細胞因子類
거세포병독감염%림파세포%Toll양수체2%Toll양수체9%수양분화인자88%세포인자류
Cytomegalovirus infections%Lymphocytes%Toll-like receptor 2%Toll-like receptor 9%Myeloid differentiation factor 88%Cytokines
目的 探讨新生小鼠巨细胞病毒(cytomegalovirus,CMV)感染后脾淋巴细胞Toll样受体(Toll like receptor,TLR)2、9,髓样分化因子88(myeloid cell differentiation 88,MyD88),白细胞介素(interleukin,IL)-2,干扰素-γ(interferon-γ,IFN-γ)和IL-10的表达变化及其在小鼠CMV感染中的作用. 方法 64只新生小鼠随机分为对照组(32只)和模型组(32只).生后4～6 h,模型组小鼠腹腔接种病毒致半数细胞感染量为104.31 U/0.1 ml的CMV悬液20μl,建立新生小鼠CMV全身播散型感染模型.2组小鼠于3、5、7、10日龄时各随机处死8只.采用聚合酶链反应技术检测7日龄小鼠肝脏、脾脏、心脏、肺脏、肾脏和唾液腺巾的CMV-DNA;采用Western印迹技术检测3、5、7、10日龄小鼠脾淋巴细胞TLR2、TLR9及MyD88蛋白的表达水平;采用酶联免疫吸附试验检测脾淋巴细胞培养上清液中IL-2、IFN-γ和IL 10水平.统计分析采用两独立样本f检验及Pearson相关分析. 结果 模型组脾淋巴细胞TLR2、TLR9和MyD88蛋白表达量在7日龄时达高峰(分别为1.06±0.09、1.19±0.03和0.89±0.07),均高于对照组(分别为0.47±0.04,0.71±0.02和0.43±0.03),差异均有统计学意义(t分别为21.69、50.40和42.06,P均＜0.01).模型组脾淋巴细胞培养上清液IL-2水平7日龄时降至最低[(16.9±7.3)pg/ml],IFNγ5日龄达高峰[(242.9±8.4)pg/ml],而IL-10在5日龄时才开始升高[(62.4±11.1) pg/ml],与正常组相应日龄时[分别为(51.9±10.8) pg/ml、(135.3±10.5) pg/ml和(36.3±5.1)pg/ml]比较,差异均有统计学意义(f分别为15.74、19.00和9.60,P均＜0.05).7日龄时,TLR2、TLR9和MyD88蛋白水平与IL2水平呈负相关(r分别为-0.978、-0.960和-0.979,P均＜0.01);与IL-10水平呈正相关(r分别为0.954、0.914和0.892,P均＜0.01). 结论 在小鼠CMV感染早期(7d内),机体可通过TLR/MyD88信号通路,激活免疫应答,在感染早期起到一定抗病毒作用.但7d后则表现为Th1/Th2细胞免疫平衡失调,使病毒易于进一步复制并导致炎症扩散.
目的 探討新生小鼠巨細胞病毒(cytomegalovirus,CMV)感染後脾淋巴細胞Toll樣受體(Toll like receptor,TLR)2、9,髓樣分化因子88(myeloid cell differentiation 88,MyD88),白細胞介素(interleukin,IL)-2,榦擾素-γ(interferon-γ,IFN-γ)和IL-10的錶達變化及其在小鼠CMV感染中的作用. 方法 64隻新生小鼠隨機分為對照組(32隻)和模型組(32隻).生後4～6 h,模型組小鼠腹腔接種病毒緻半數細胞感染量為104.31 U/0.1 ml的CMV懸液20μl,建立新生小鼠CMV全身播散型感染模型.2組小鼠于3、5、7、10日齡時各隨機處死8隻.採用聚閤酶鏈反應技術檢測7日齡小鼠肝髒、脾髒、心髒、肺髒、腎髒和唾液腺巾的CMV-DNA;採用Western印跡技術檢測3、5、7、10日齡小鼠脾淋巴細胞TLR2、TLR9及MyD88蛋白的錶達水平;採用酶聯免疫吸附試驗檢測脾淋巴細胞培養上清液中IL-2、IFN-γ和IL 10水平.統計分析採用兩獨立樣本f檢驗及Pearson相關分析. 結果 模型組脾淋巴細胞TLR2、TLR9和MyD88蛋白錶達量在7日齡時達高峰(分彆為1.06±0.09、1.19±0.03和0.89±0.07),均高于對照組(分彆為0.47±0.04,0.71±0.02和0.43±0.03),差異均有統計學意義(t分彆為21.69、50.40和42.06,P均＜0.01).模型組脾淋巴細胞培養上清液IL-2水平7日齡時降至最低[(16.9±7.3)pg/ml],IFNγ5日齡達高峰[(242.9±8.4)pg/ml],而IL-10在5日齡時纔開始升高[(62.4±11.1) pg/ml],與正常組相應日齡時[分彆為(51.9±10.8) pg/ml、(135.3±10.5) pg/ml和(36.3±5.1)pg/ml]比較,差異均有統計學意義(f分彆為15.74、19.00和9.60,P均＜0.05).7日齡時,TLR2、TLR9和MyD88蛋白水平與IL2水平呈負相關(r分彆為-0.978、-0.960和-0.979,P均＜0.01);與IL-10水平呈正相關(r分彆為0.954、0.914和0.892,P均＜0.01). 結論 在小鼠CMV感染早期(7d內),機體可通過TLR/MyD88信號通路,激活免疫應答,在感染早期起到一定抗病毒作用.但7d後則錶現為Th1/Th2細胞免疫平衡失調,使病毒易于進一步複製併導緻炎癥擴散.
목적 탐토신생소서거세포병독(cytomegalovirus,CMV)감염후비림파세포Toll양수체(Toll like receptor,TLR)2、9,수양분화인자88(myeloid cell differentiation 88,MyD88),백세포개소(interleukin,IL)-2,간우소-γ(interferon-γ,IFN-γ)화IL-10적표체변화급기재소서CMV감염중적작용. 방법 64지신생소서수궤분위대조조(32지)화모형조(32지).생후4～6 h,모형조소서복강접충병독치반수세포감염량위104.31 U/0.1 ml적CMV현액20μl,건립신생소서CMV전신파산형감염모형.2조소서우3、5、7、10일령시각수궤처사8지.채용취합매련반응기술검측7일령소서간장、비장、심장、폐장、신장화타액선건적CMV-DNA;채용Western인적기술검측3、5、7、10일령소서비림파세포TLR2、TLR9급MyD88단백적표체수평;채용매련면역흡부시험검측비림파세포배양상청액중IL-2、IFN-γ화IL 10수평.통계분석채용량독립양본f검험급Pearson상관분석. 결과 모형조비림파세포TLR2、TLR9화MyD88단백표체량재7일령시체고봉(분별위1.06±0.09、1.19±0.03화0.89±0.07),균고우대조조(분별위0.47±0.04,0.71±0.02화0.43±0.03),차이균유통계학의의(t분별위21.69、50.40화42.06,P균＜0.01).모형조비림파세포배양상청액IL-2수평7일령시강지최저[(16.9±7.3)pg/ml],IFNγ5일령체고봉[(242.9±8.4)pg/ml],이IL-10재5일령시재개시승고[(62.4±11.1) pg/ml],여정상조상응일령시[분별위(51.9±10.8) pg/ml、(135.3±10.5) pg/ml화(36.3±5.1)pg/ml]비교,차이균유통계학의의(f분별위15.74、19.00화9.60,P균＜0.05).7일령시,TLR2、TLR9화MyD88단백수평여IL2수평정부상관(r분별위-0.978、-0.960화-0.979,P균＜0.01);여IL-10수평정정상관(r분별위0.954、0.914화0.892,P균＜0.01). 결론 재소서CMV감염조기(7d내),궤체가통과TLR/MyD88신호통로,격활면역응답,재감염조기기도일정항병독작용.단7d후칙표현위Th1/Th2세포면역평형실조,사병독역우진일보복제병도치염증확산.
Objective To investigate the alteration of Toll-like receptor (TLR) 2 and 9,myeloid cell differentiation 88 (MyD8),interleukin-2 (IL 2),interferon-γ(IFN γ) and interleukin-10 (IL-10) expression in spleen lymphocytes of cytomegalovirus (CMV) infected newborn murine.Methods Sixty-four neonatal mice were randomly divided into control group and CMV infected group (32 mice in each).Mice in CMV infected group were injected intraperitoneally with 20 μl CMV suspension (tissue culture infections dose 50 of 104.31 U/0.1 ml) at 4 6 hours after birth to create the systematic CMV infection models.At 3,5,7 and 10 days old,eight mice randomly selected from each group were sacrificed.Polymerase chain reaction was applied to detect CMV-DNA in liver,spleen,heart,lung,kidney and salivary glands of 7 day old mice.Western blot was used to measure the protein expression of T1R 2,9 and MyD88 in spleen lymphocyte,and enzyme linked immunosorbent assay was used to test the levels of IL-2,IFN-γand IL-10 in supernatant of spleen lymphocyte from 3,5,7 and 10-dayold mice.Independent samples t test and Pearson correlation analysis were applied as the statistical methods.Results On the 7th day after CMV infection,the protein expression of TLR 2,9 and MyD88 in spleen lymphocyte in CMV infected group increased to the maximum level (1.06±0.09,1.19±0.03 and 0.89±0.07),which were higher than those in control group (0.47±0.04,0.71 ±0.02 and 0.43±0.03),the differences were statistically significant(t=21.69,50.40 and 42.06,P＜0.01,respectively).In CMV infected group,expression of IL-2 decreased to the lowest level on the 7th day after CMV infection [(16.9±7.3) pg/ml],IFN γ reached its peak level on the 5th day [(242.9±8.4) pg/ml] and IL-10 increased since the 5th day [(62.4±11.1) pg/ml],there were statistically significant differences as compared with control group [(51.9± 10.8) pg/ml,(135.3±10.5) pg/ml and (36.3±5.1) pg/ml](t=15.74,19.00 and 9.60,P＜0.05,respectively).There was a negative correlation between TLR2,TLR9,MyD88 protein expression and IL-2 level (r =0.978,0.960 and-0.979,all P＜0.01,respectively) and a positive correlation between TLR2,TLR9,MyD88 protein and IL-10 level (r =0.954,0.914 and 0.892,P ＜ 0.01,respectively).Conclusions In the early phase of CMV infection (within 7 days),immunity is activated through TLR/MyD88 signal pathway resulting anti-infection effect.However,after the 7th day,Th1/Th2 cellular immunity imbalance allows virus to replicate easily.
