中华妇产科杂志
中華婦產科雜誌
중화부산과잡지
CHINESE JOUNAL OF OBSTETRICS AND GYNECOLOGY
2013年
6期
453-456
,共4页
贺芳%陈敦金%李连英%陈启宪
賀芳%陳敦金%李連英%陳啟憲
하방%진돈금%리련영%진계헌
妊娠中期%活性氧%一氧化氮%细胞因子类%淋巴细胞活化
妊娠中期%活性氧%一氧化氮%細胞因子類%淋巴細胞活化
임신중기%활성양%일양화담%세포인자류%림파세포활화
Pregnancy trimester,second%Reactive oxygen species%Nitric oxide%Cytokines%Lymphocyte activation
目的 探讨妊娠对小鼠外周血中性粒细胞(PMN)胞内活性氧簇(ROS)、血清NO和细胞因子水平、T淋巴细胞活化的影响.方法 以处于中孕期(孕14 d)的BALB/c小鼠为研究对象,分为妊娠组(为正常孕鼠)、对照组(为正常非孕雌鼠).以2,7-二氯双氢荧光素乙二脂(H2DCFDA)进行外周血染色,结合流式细胞仪检测外周血PMN胞内ROS水平；Griess试剂盒检测外周血血清中NO的水平；用流式微球阵列(CBA)方法检测血清细胞因子白细胞介素(IL)6、IL-10、单核细胞趋化蛋白l(MCP-1)、干扰素γ(IFN-γ)、肿瘤坏死因子α(TNF-α)和IL-12的水平；采用荧光抗体双色标记方法结合流式细胞仪分别检测外周血T淋巴细胞早、中、晚期活化标志——CD69、CD25及CD71的表达率.结果 与对照组小鼠外周血PMN胞内ROS水平(101.1 ±2.2)比较,妊娠组小鼠ROS水平升高(为134.5 ±10.3),两组比较,差异有统计学意义(P＜0.05).对照组小鼠外周血血清NO水平为(22.7±0.7) μmol/L,妊娠组小鼠血清NO水平提高到(36.3±1.2)μmol/L,两组比较,差异有统计学意义(P＜0.01).对照组小鼠外周血IL-6、IL-10、MCP-1、IFN-γ、TNF-α和IL-12 6种细胞因子的水平分别为(9.3±0.5)、(26.7±0.9)、(21.2±1.6)、(14.5±1.8)、(22.6±1.6)和(8.4±1.2)pg/ml,妊娠组分别为(26.5±1.0)、(40.4±2.5)、(25.1±0.7)、(457.4±17.9)、(93.2±4.3)及(7.5±0.9) pg/ml；与对照组分别比较,妊娠组小鼠外周血血清中IL-6、IFN-γ、TNF-α的水平显著升高(P＜0.01),但IL-10、MCP-1的水平则无差异(P＞0.05).对照组小鼠外周血CD3+ T淋巴细胞中CD69、CD25和CD71的表达率分别为(0.43±0.15)％、(5.13±0.25)％和(0.37±0.11)％,而妊娠组分别为(0.40±0.10)％、(6.17±0.40)％及(6.10±0.31)％；与对照组分别比较,CD25和CD71的表达率明显升高(P＜0.05),CD69则无明显差异(P=0.15).结论 中孕期小鼠外周血PMN胞内ROS水平和血清NO水平升高,细胞因子IL-6、IFN-γ、TNF-α水平增加,T淋巴细胞的中、晚期活化能力增强.
目的 探討妊娠對小鼠外週血中性粒細胞(PMN)胞內活性氧簇(ROS)、血清NO和細胞因子水平、T淋巴細胞活化的影響.方法 以處于中孕期(孕14 d)的BALB/c小鼠為研究對象,分為妊娠組(為正常孕鼠)、對照組(為正常非孕雌鼠).以2,7-二氯雙氫熒光素乙二脂(H2DCFDA)進行外週血染色,結閤流式細胞儀檢測外週血PMN胞內ROS水平；Griess試劑盒檢測外週血血清中NO的水平；用流式微毬陣列(CBA)方法檢測血清細胞因子白細胞介素(IL)6、IL-10、單覈細胞趨化蛋白l(MCP-1)、榦擾素γ(IFN-γ)、腫瘤壞死因子α(TNF-α)和IL-12的水平；採用熒光抗體雙色標記方法結閤流式細胞儀分彆檢測外週血T淋巴細胞早、中、晚期活化標誌——CD69、CD25及CD71的錶達率.結果 與對照組小鼠外週血PMN胞內ROS水平(101.1 ±2.2)比較,妊娠組小鼠ROS水平升高(為134.5 ±10.3),兩組比較,差異有統計學意義(P＜0.05).對照組小鼠外週血血清NO水平為(22.7±0.7) μmol/L,妊娠組小鼠血清NO水平提高到(36.3±1.2)μmol/L,兩組比較,差異有統計學意義(P＜0.01).對照組小鼠外週血IL-6、IL-10、MCP-1、IFN-γ、TNF-α和IL-12 6種細胞因子的水平分彆為(9.3±0.5)、(26.7±0.9)、(21.2±1.6)、(14.5±1.8)、(22.6±1.6)和(8.4±1.2)pg/ml,妊娠組分彆為(26.5±1.0)、(40.4±2.5)、(25.1±0.7)、(457.4±17.9)、(93.2±4.3)及(7.5±0.9) pg/ml；與對照組分彆比較,妊娠組小鼠外週血血清中IL-6、IFN-γ、TNF-α的水平顯著升高(P＜0.01),但IL-10、MCP-1的水平則無差異(P＞0.05).對照組小鼠外週血CD3+ T淋巴細胞中CD69、CD25和CD71的錶達率分彆為(0.43±0.15)％、(5.13±0.25)％和(0.37±0.11)％,而妊娠組分彆為(0.40±0.10)％、(6.17±0.40)％及(6.10±0.31)％；與對照組分彆比較,CD25和CD71的錶達率明顯升高(P＜0.05),CD69則無明顯差異(P=0.15).結論 中孕期小鼠外週血PMN胞內ROS水平和血清NO水平升高,細胞因子IL-6、IFN-γ、TNF-α水平增加,T淋巴細胞的中、晚期活化能力增彊.
목적 탐토임신대소서외주혈중성립세포(PMN)포내활성양족(ROS)、혈청NO화세포인자수평、T림파세포활화적영향.방법 이처우중잉기(잉14 d)적BALB/c소서위연구대상,분위임신조(위정상잉서)、대조조(위정상비잉자서).이2,7-이록쌍경형광소을이지(H2DCFDA)진행외주혈염색,결합류식세포의검측외주혈PMN포내ROS수평；Griess시제합검측외주혈혈청중NO적수평；용류식미구진렬(CBA)방법검측혈청세포인자백세포개소(IL)6、IL-10、단핵세포추화단백l(MCP-1)、간우소γ(IFN-γ)、종류배사인자α(TNF-α)화IL-12적수평；채용형광항체쌍색표기방법결합류식세포의분별검측외주혈T림파세포조、중、만기활화표지——CD69、CD25급CD71적표체솔.결과 여대조조소서외주혈PMN포내ROS수평(101.1 ±2.2)비교,임신조소서ROS수평승고(위134.5 ±10.3),량조비교,차이유통계학의의(P＜0.05).대조조소서외주혈혈청NO수평위(22.7±0.7) μmol/L,임신조소서혈청NO수평제고도(36.3±1.2)μmol/L,량조비교,차이유통계학의의(P＜0.01).대조조소서외주혈IL-6、IL-10、MCP-1、IFN-γ、TNF-α화IL-12 6충세포인자적수평분별위(9.3±0.5)、(26.7±0.9)、(21.2±1.6)、(14.5±1.8)、(22.6±1.6)화(8.4±1.2)pg/ml,임신조분별위(26.5±1.0)、(40.4±2.5)、(25.1±0.7)、(457.4±17.9)、(93.2±4.3)급(7.5±0.9) pg/ml；여대조조분별비교,임신조소서외주혈혈청중IL-6、IFN-γ、TNF-α적수평현저승고(P＜0.01),단IL-10、MCP-1적수평칙무차이(P＞0.05).대조조소서외주혈CD3+ T림파세포중CD69、CD25화CD71적표체솔분별위(0.43±0.15)％、(5.13±0.25)％화(0.37±0.11)％,이임신조분별위(0.40±0.10)％、(6.17±0.40)％급(6.10±0.31)％；여대조조분별비교,CD25화CD71적표체솔명현승고(P＜0.05),CD69칙무명현차이(P=0.15).결론 중잉기소서외주혈PMN포내ROS수평화혈청NO수평승고,세포인자IL-6、IFN-γ、TNF-α수평증가,T림파세포적중、만기활화능력증강.
Objective To investigate the influence of pregnancy on the production of reactive oxygen species (ROS) from mouse peripheral blood neutrophils (PMN),the levels of NO and cytokines from serum,the activation of T lymphocytes,and initially find the immune regulation effects of pregnancy on the mouse peripheral blood lymphocytes.Methods Take the BALB/c mice which were at the mid trimester of pregnancy (day 14) as the object,full blood staining using ROS probe 2',7'-dichlorodihydrofluorescein diacetate (H2DCFDA) combing with flow cytometry was used to test the levels of ROS from PMN.The production of NO from peripheral blood serum were analyzed by Griess kit while the soluble cytokines interleukin (IL) 6,IL-10,monocyte chemotactic protein 1 (MCP-1),interferon γ (IFN-γ),tumor necrosis factor α (TNF-α) and IL-12 were detected by liquid protein quantitative technology cytometric bead array (CBA) using flow cytometry.The activation of peripheral blood lymphocytes at early,middle and later phases which marked with CD69,CD25 and CD71 respectively were tested by flow cytometry and two-color fluorescent staining.Results Comparing to the normal non-pregnant mouse,pregnancy obviously promoted the production of ROS from PMN (101.1 ± 2.2 versus 134.5 ± 10.3,P ＜ 0.05).Comparing to the normal non-pregnant mouse,pregnancy obviously promoted the secretion of NO [(22.7 ± 0.7) versus (36.3 ±1.2) μmol/L,P ＜0.01].In normal non-pregnant mouse,the serum levels of IL-6,IL-10,MCP-1,IFN-γ,TNF-αandlL-12were (9.3±0.5),(26.7±0.9),(21.2±1.6),(14.5 ±1.8),(22.6±1.6) and (8.4 ± 1.2) pg/ml,while in pregnancy group the levels were (26.5 ± 1.0),(40.4 ± 2.5),(25.1 ±0.7),(457.4 ± 17.9),(93.2 ± 4.3) and (7.5 ± 0.9) pg/ml correspondingly ; the levels of IL-6,IFN-γ,TNF-α from peripheral blood serum (P ＜ 0.01),while had no effects on the production of IL-10 and MCP-1 (P ＞ 0.05).About the CD3+ T lymphocytes activation,in normal non-pregnant mouse,the CD69,CD25 and CD71 expression rate were (0.43 ±0.15)％,(5.13 ±0.25)％ and (0.37 ±0.11)％,while in pregnancy group the CD69,CD25 and CD71 expression rate were (0.40 ±0.10)％,(6.17 ±0.40)％ and (6.10 ±0.31)％.The levels of middle and later phases markers as CD25 and CD71 were highly up-regulated (P ＜ 0.05),while the early phase action CD69 had no obvious variation (P ＞ 0.05).Conclusion The mid trimester of pregnancy promoted the production of ROS from PMN,the levels of NO,IL-6,IFN-γ,TNF-αfrom peripheral blood serum,and the middle-and later-phase activation of T lymphocytes.
