The Role Of TLR3 And Placenta Apoptosis In Placenta HBV Infection And HBV Intrauterine Infection

OBJECTIVE:①To show the condition of apoptosis in term human placenta of HBsAg-positive pregnant women.③To explore the relationship between placenta apoptosis, Toll-Like Receptor 3 and placenta HBV infection.③To explore the effection of TLR3 and placenta apoptosis in HBV intrauterine affection.METHODS:1. From July 2003 to June 2004,136 HBsAg-positive pregnant women were collected in Taiyuan infectious hospital and 30 normal pregnant women were collected in NO.1 hospital attached to shanxi Medical University. Right after delivery maternal elbow vein blood and newborn femoral vein blood were collected. At the same time, the placental tissues were also collected. The epidemiological base line data involving gestation and postpartum were also collected. Then the following studies were performed:①HBsAg in serum were tested by ELISA; HBV DNA in serum was detected by fluorescent quantitative PCR.②Both HBsAg and TLR3 in infected placental tissues were detected by double labeling immunofluorescent histochemistry assay.③The condition of apoptosis in placenta were detected by TdT-mediated dUTP nick end labeling assay (Tunel).2. From December 2008 to January 2009,20 HBsAg-positive pregnant women who were defined HBV intrauterine infected were collected in Taiyuan infectious hospital,60 HBsAg-positive but HBV intrauterine non-infected pregnant women and 40 normal pregnant women were also collected by ratio 1:3 and 1:2 separately. Fresh placenta were collected sterility to extract total RNA,2ml maternal elbow vein blood and 2ml newborn femoral vein blood were collected. Then the following studies were performed:①HBsAg in serum were tested by ELISA; HBV DNA in serum was detected by fluorescent quantitative PCR.②TLR3 mRNA and caspase3 mRNA expression were detection by Real-time RT-PCR.RESULTS:①Compare the index of apoptosis (IA) by groups:the IA of HBsAg-positive pregnant women was higher than normal pregnant women (t=37.834, P<0.001),but no significant difference among each HBV amount group. The IA of placenta HBV infected women was significant lower than placenta HBV non-infected women (t=2.614, p=0.010). The IA was no significant difference between of placenta TLR3-positive women and placenta TLR3-negative women or between HBV intrauterine infected and non-infected women.②Compare TLR3 mRNA and caspase3 mRNA in placenta by groups. The relative expression of TLR3 mRNA in HBsAg-positive pregnant women was higher than normal pregnant women (t=-21.714, P<0.0001), but there was not significant difference among each HBV amount group or between HBV intrauterine infected and non-infected women. The relative expression of caspase3 mRNA in HBsAg-positive pregnant women was higher than normal pregnant women (t=-2.467,P=0.015), but there was also not significant difference among each HBV amount group or between HBV intrauterine infected and non-infected women.③In normal pregnant women, there was negative correlation of Caspas3_Ct/GAPDH Ct and TLR3_Ct/β-actin Ct(r=-0.479,p=0.002),while there was no correlation of them in HBsAg-positive pregnant women, whether HBV intrauterine infected or not.CONCLUSIONS:①Both the IA and caspase3 mRNA in placenta of normal pregnant women were higher than HBsAg-positive pregnant women, indicate that HBV could increase the apoptosis of placenta.②In HBsAg-positive pregnant women, the IA of placenta HBV infected women was significant lower than placenta HBV non-infected women, indicate that the apoptosis of placenta may be a protective factor against placenta HBV infection.③The expression of TLR3 mRNA in normal pregnant women's placenta was significant lower than HBsAg-positive pregnant women, imply that HBV may up-regulation TLR3 of placenta。④The correlation of TLR3 and placenta apoptosis in HBsAg-positive pregnant women has not been found yet.