新生小鼠巨细胞病毒感染早期脾淋巴细胞Toll样受体2和9与髓样分化因子88及Th1／Th2类细胞因子的表达变化  

Expression of Toll-like receptor 2 and 9, myeioid cell differentiation 88 and Th1/Th2 eytokines in spleen lymphocytes of early cytomegalovirus infected newborn murine

作　　者：郑玉艳[1] 王军[1] 叶黎离[1] 

机构地区：[1]徐州医学院附属医院儿科,221002

出　　处：《中华围产医学杂志》2013年第11期677-681,共5页Chinese Journal of Perinatal Medicine

基　　金：江苏省“六大人才高峰计划”项目（2011WS-075）

摘　　要：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4～6h，模型组小鼠腹腔接种病毒致半数细胞感染量为104．31U／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、肾脏和唾液腺中的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t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]，IFN-γ5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（t分别为-15．74、19．00和9．60，P均〈0．05）。7日龄时，TILR2、TLR9和MyD88蛋白水平与IL-2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〈0．01Objective To investigate the alteration of Toll-like receptor （TLR） 2 and 9, myeloid cell differentiation 88 （ MyDS）, interleukin-2 （ IL-2 ）, interferon-γ（ IFN-γ） and interleukin 10 （IL-10） expression in spleen Iymphocytes 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 20μl 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 TLR 2, 9 and MyD88 in spleen lymphocyte, and enzyme linked immunosorbent assay wits used to test the levels of IL-2, IFN γ and IL-10 in supernalant of spleen lymphocyte from 3,5,7 and 10 day old 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. 7100. 02 and 0. 4±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 CMVinfection [（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]�

关 键 词：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 

分 类 号：R722.1[医药卫生—儿科]