TLR4, MD2 And CD14 Mediated IL-6 And TNF-α In The Onset Of Preterm Delivery

Objective:To study the expression of TLR4,MD2,CD14mRNA in peripheral blood mononuclear cells of patients of preterm labor(in vaginal), normal term labor(in vaginal), and normal term labor(in cesarean section delivery without labor).To Analyze the relationship of TLR4,MD2 and CD 14 expression with premature delivery and its effect on the pathogenesis of subclinical chorioamnionitis.Methods:To chose 20 cases of preterm birth patients in labor,28 of cases of normal term birth patients in labor,10 cases of normal term birth with caesarean delivery before threatened labor. To observe the expression of the TLR4, MD2,CD14mRNA in peripheral blood mononuclear cells by reverse transcription reaction (RT-PCR).And the change of placenta and fetal membrane with subclinical chorioamnionitis by HE staining.Result:1.The expression of TLR4mRNA(0.41±0.17) and MD2mRNA (0.39±0.20) in preterm birth group were significantly higher than normal term birth patients in labor group TLR4mRNA(0.18±0.85),MD2mRNA (0.22±0.13)(p<0.05),but there is no difference for the CD14mRNA in the two groups(1.82±0.58 Vs 1.69±0.53)(p=0.42>0.05),all of there are high expression; the expression of TLR4,MD2,CD14mRNA in the two groups are higher than the normal term birth with caesarean delivery group,but there is no statistic difference between the two term groups(p >0.05).2.There were 14 cases who were subclinical chorioamnionitis patients in 20 cases of preterm birth patients in labor(70%).6 cases are mild.5 cases are midrange,3 cases are severe among them.There were 9 cases who were subclinical chorioamnionitis patient in 28 cases of the normal term group (32.14%).5 cases are mild,3 cases are midrange,1 is severe among them.There were 2 cases who were mild subclinical chorioamnionitis patient in 10 cases of the normal term birth with caesarean delivery group.The expression of TLR4,MD2 and CD14mRNA in subclinical chorioamnionitis group were significantly higher than without subclinical chorioamnionitis group.To compare the correlation of TLR4,MD2 and CD14mRNA and the chorioamnionitis, r are 0.635,0.467,0.404,p<0.05.3.There are positive correlation between the expression of TLR4,MD2 and CD14mRNA in the maternal peripheral blood.Conclusion:1.The expression of TLR4, MD2 coincidenced with pregnant weeks and the chorioamnionitis condition, without the modus of labor.2.The expression of CD14 in two vaginal group are higher than the normal term birth with caesarean delivery group, is coincidenced with the modus of labor and the chorioamnionitis condition.Objective:To study the expression and the relation between TLR4. MD2 and CD14 in the placenta.Methods:The expression of TLR4, MD2 and CD 14 in placenta were detected by immunohistochemical method in the three groups.Result:1.Immunoreactivity for TLR4 was found in the cytoplasm of the syncytiotrophoblast, with darker staining in some areas of the maternal facing plasma membrane (MVM),and was also found in the cytoplasm of the extravillous trophoblast (EVT) and the amniotic membrane, the nucleus is negative. MD2 was found in the cytoplasm of the syncytiotrophoblast, EVT, the hofbauer cell and the amniotic membrane, the nucleus is strong positive. CD 14 was found in the cytoplasm of the syncytiotrophoblast and the amniotic membrane, EVT is negative.2.Immunoreactivity for TLR4, MD2 and CD14 is statistic difference between the three groups (p<0.05).The expression of TLR4, MD2 and CD 14 in preterm labor group were significantly higher than the normal term birth patients in labor group, the two group all significantly higher than the normal term birth with caesarean delivery group.3.Immunoreactivity for TLR4, MD2 and CD 14 in subclinical chorioamnionitis group were significantly higher than without subclinical chorioamnionitis group.There is statistic difference between the two groups (p<0.05).4. There are positive correlation between the expression of TLR4, MD2 and CD 14 in the maternal-fetal interface.Conclusion:1.The syncytiotrophoblast and amniotic membrane express the TLR4, MD2 and CD14, but the EVT just express the TLR4, MD2.2.The expression of TLR4, MD2 and CD 14 in the placenta coincidenced with pregnant weeks, the modus of labor and the chorioamnionitis condition.Objective:The aim of the study was to investigate the expression of TLR4, MD2 and CD 14 in primary chorionic trophocyte cell with or without the stimulation of LPS,the effect of TLR4, MD2 and CD 14 on the cell and the secretion of IL-6 and TNF-a induced by LPS. Methods:1.Trophoblasts were isolated from chorionic of placenta and cultured in vitro.Cultured chorionic trophocyte cells were treated with LPS for different time (0 h,2 h,4 h,8 h,12 h,24 h) and different concentration. The expression of TLR4 mRNA was detected by reverse transcription polymerase chain reaction (RT-PCR), and the secretion of IL-6 and TNF-a was measured by enzyme linked immunosorbent assay (ELISA).To known the efficiency concentration and the efficiency time.2.Pretreated with TLR4Ab, to investigate the change of TLR4, MD2 and CD14mRNA and protein, the secretion of IL-6 and TNF-a induced by LPS.Results:1.TLR4,MD2 and CD 14 mRNA was expressed in primary chorionic trophoblast cells and the expression of mRNA increased when cells treated whith LPS.2. LPS could increase the secretion of IL-6 and TNF-a in a dose-dependent and time-dependent manner, the efficiency concentration is 10ng/ml and the efficiency time is 12h3.TLR4, MD2, CD 14 mRNA and protein was increase after treated whith LPS,could also be blocked by TLR4 Ab.Conclusion:1.Cultured primary chorionic trophocytes express TLR4,MD2, CD14 mRNA.TLR4-MD2-CD14 may play an important role in LPS-induced secretion of IL-6 and TNF-a. TLR4 Ab could block this effect.2.There is a immune tolerance phenomenon in trophocytes cell.3.LPS-TLR4-MD2-CD14-proinflammation cytokines may take part in the inflammation reaction of trophoblast cells which induced preterm labor.