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目的检测正常孕妇妊娠不同阶段外周血单个核细胞(PBMC)巨噬细胞移动因子(MIF)基因表达水平及血清中MIF含量的变化,为揭示妊娠期间的调控机制提供实验依据。方法 42名妊娠不同阶段的正常孕妇来自天津市中医药大学第一附属医院,年龄26～36岁,平均(31.0±3.0)岁,其中妊娠早期18例,孕龄(8.71±1.35)周;妊娠中期9例,孕龄(21.35±3.37)周;妊娠晚期15例,孕龄(33.01±2.32)周。9例非妊娠对照组来自天津市血液中心健康女性志愿者,年龄(26.5±3.0)岁。用密度梯度离心法分离正常孕妇PBMC,调整细胞浓度为1×106个,提取RNA,反转录(RT)合成cDNA,用荧光定量聚合酶链反应(PCR)方法检测MIFmRNA表达水平,用酶联免疫吸附(ELISA)法检测孕妇血清中MIF含量。结果妊娠早期、中期、晚期PBMCMIF基因表达水平分别为2.15±0.47、2.07±0.39、2.18±0.54,与非妊娠对照组(0.68±0.12)比较,均有明显升高,差异有统计学意义(P<0.05)。妊娠早期、中期、晚期血清中MIF含量分别为(15.25±1.72)、(16.12±1.99)、(15.55±1.82)ng/ml,与非妊娠对照组([5.53±1.77)ng/ml]比较,均有明显升高,差异有统计学意义(P<0.05)。不同妊娠时间比较,MIFmRNA表达水平和MIF含量的差异均无统计学意义(P>0.05)。结论 MIF的基因表达和含量在整个妊娠过程中都维持在较高水平,提示MIF参与了整个妊娠至分娩。
Objective To detect the expression of MIF gene in peripheral blood mononuclear cells (PBMC) and the content of MIF in normal pregnant women at different stages of pregnancy, and provide experimental evidence for revealing the regulation mechanism during pregnancy. Methods Forty-two pregnant women of different stages of pregnancy were from the First Affiliated Hospital of Tianjin University of Traditional Chinese Medicine, aged from 26 to 36 years, with an average of (31.0 ± 3.0) years. Among them, 18 were pregnant during the first trimester and 8.71 ± 1.35 weeks There were 9 cases in the middle period and 21 weeks in the gestational age group (21.35 ± 3.37) weeks, 15 cases in the third trimester of pregnancy and the gestational age (33.01 ± 2.32) weeks. Nine non-pregnant controls were from healthy female volunteers in Tianjin Blood Center, aged (26.5 ± 3.0) years. The PBMC of normal pregnant women were separated by density gradient centrifugation, the concentration of cells was adjusted to 1 × 106, RNA was extracted and reverse transcription (RT) was used to synthesize cDNA. The expression of MIF mRNA was detected by fluorescence quantitative polymerase chain reaction (PCR) MIF content in serum of pregnant women was detected by ELISA. Results The expression levels of PBMCMIF gene in early, middle and late pregnancy were 2.15 ± 0.47, 2.07 ± 0.39 and 2.18 ± 0.54, respectively, which were significantly higher than those in non-pregnant control group (0.68 ± 0.12) (P <0.05). The levels of MIF in the early, middle and late pregnancy were (15.25 ± 1.72), (16.12 ± 1.99) and (15.55 ± 1.82) ng / ml, respectively, compared with those in the non-pregnant control group (5.53 ± 1.77 ng / ml) Were significantly increased, the difference was statistically significant (P <0.05). There was no significant difference in MIF mRNA expression and MIF content between different pregnancy stages (P> 0.05). Conclusion MIF gene expression and content are maintained at a high level throughout pregnancy, suggesting that MIF is involved in the entire pregnancy to childbirth.