Relationship Between TLRs Expression In Placenta And Hepatitis B Virus Infection Of Pregnant Women

OBJECTIVE:①To show the changes of TLR3 mRNA,TLR4 mRNA,TLR7 mRNA and TLR9 mRNA expression in term human placenta of normal and HBsAg-positive pregnant women.②To explore the distribution of Toll-Like Receptor 3 in term human placenta of normal and HBsAg-positive pregnant women.③To show the changes of TLR3 protein expression in term human placenta of normal and HBsAg-positive pregnant women.METHODS: From July 2006 to Octomber 2007 ,116 HBsAg-positive pregnant women were collected in Taiyuan infectious hospital and 41 normal pregnant women were collected in NO.1 hospital of shanxi Medical University. The epidemiological base line data involving gestation and postpartum were also collected. Maternal elbow vein blood were collected. At the same time, the placental tissues were also collected. Following studies were performed: HBsAg in gravidas'serum were tested by ELISA; HBV DNA was detected by fluorescent quantitative PCR TLR3 mRNA,TLR4 mRNA,TLR7 mRNA and TLR9 mRNA expression in term human placenta of normal and HBsAg-positive pregnant women were detection by RT-PCR;TLR3 in placentas were detected by immunohistochemistry ABC; TLR3 protein expression in term human placenta of normal and HBsAg-positive pregnant women were detection by western-blot; All of the data was analyzed by the statistical packages Visual Fox Por 6.0, SPSS 13.0 for windows.RESULTS:①TLR3 mRNA,TLR4 mRNA,TLR7 mRNA and TLR9 mRNA were found in human term placenta of normal and HBsAg-positive pregnant women,RT-PCR revealed that TLR3 decreased in HBV infection women's plancenta(t=3.003,P<0.05)and there was a significant increase in mRNA expression in human term placenta of pregnancy complicated with HBV for TLR4,TLR7 and TLR9 (t=3.530,t=4.036,t=3.3834, P<0.05). There was no significant difference in mRNA expression in human term placenta of pregnancy complicated with HBV for TLR4,TLR7 and TLR9 (t =4.60,t =3.50,t =1.236, P>0.05).②The positive rate of TLR3 in HBsAg-positive pregnant women was 73.28%(85/116). TLR3 was mostly located in the cytoplasm, a part on the cell membrane and occasionally in the nuceus. TLR3 positive rate gradually increased from the maternal side to the fetal side (trend test:χ2=61.654,P=0.000) in the placentalcell layers. Maternal decidual cells (30.17%) < trophoblastic cells (45.69%) < villous mesenchymal cells(70.69%) < villous capillary endothelial cells (73.28%). There was statistical difference in distribution of TLR3 between placentas of normal and HBsAg-positive pregnant women (χ2=12.018,P=0.001) and There was statistical differenceof the rate of intrauterine infection because of the distribution of TLR3 in placental barrier (χ2=5.769,P=0.016).③TLR3 protein was found in human term placenta of normal and HBsAg-positive pregnant women,western-blot revealed that there was a significant decrease in protein expression in human term placenta of pregnancy complicated with HBV for TLR3 (t=5.183,P=0.000).CONCLUSIONS:①TLR3 was mostly located in the cytoplasm ,TLR3 was expressed in different layers of placental cells,TLR3 positive rate gradually increased from the maternal side to the fetal side. TLR3 mRNA and TLR3 protein was found in human term placenta of normal and HBsAg-positive pregnant women,there was a significant decrease in protein and gene expression in human term placenta of pregnancy complicated with HBV for TLR3,The down-regulation of TLR3 associates with HBV infection in pregnant women ,TLR3 perhaps can effectively protect placenta from HBV infection and intrauterine infection.②TLR4 mRNA,TLR7 mRNA and TLR9 mRNA were found in human term placenta of normal and HBsAg-positive pregnant women,there was a significant increase in gene expression in human term placenta of pregnancy complicated with HBV for TLR4,TLR7 and TLR9.All these indicate that TLRs play an important roles in HBV infects pregnant women's placenta...