Effects of pregnancy on the ROS, NO, cytokine levels and lymphocytes activation from mouse peripheral blood
10.3760/cma.j.issn.0529-567x.2013.06.011
- VernacularTitle:妊娠对小鼠外周血ROS、NO、细胞因子水平及T淋巴细胞活化的影响
- Author:
Fang HE
;
Dunjin CHEN
;
Lianying LI
;
Qixian CHEN
- Publication Type:Journal Article
- Keywords:
Pregnancy trimester,second;
Reactive oxygen species;
Nitric oxide;
Cytokines;
Lymphocyte activation
- From:
Chinese Journal of Obstetrics and Gynecology
2013;(6):453-456
- CountryChina
- Language:Chinese
-
Abstract:
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.
