| Objective:To preliminary explore the mechanism of adverse pregnancy outcomes caused by infection with Chinese 1 Toxoplasma.Methods:Animal models of C57BL/6 mice infected by WH3 and WH6 strains of T.gondii in early pregnancy were established.Severity of adverse pregnancy outcomes The statistics of abortion rates and the HE staining of the decidua tissue were used to evaluate the severity of adverse pregnancy outcomes.Single cell suspensions of uterine tissues from the mice infected with different strains of Toxoplasma in the early and second trimester of pregnancy were analyzed by flow cytometry to detect the changes of decidual tissue resident NK(CD49a~+Eomes~+tr NK)cells and the changes in secretion of GPFs:pleiotrophin(PTN),osteopontin(OPN)and osteoglycin(OGN).The changes of the GPFs were verified by Western blotting.To observe the improvement of the developmental restriction caused by GPFs,PTN,OPN and OGN were administered to the pregnant mice intravenously in the first trimester,respectively,and then immunohistochemistry ofα-SMA in the uterine tissue of mice at gd 9 and immunohistochemistry of Neu N in the embryonic tissue at gd 15 were conducted,respectively.HE staining of fetal mouse eye tissues of gd 15 was performed to observe the effect of supplementation of GPFs on the development of fetal mouse eye tissues;Alcian blue-alizarin red staining was used to evaluate the skeletal tissue development of fetal mice of 15 days of pregnancy after supplementing GPFs.Results:(1)Adverse pregnancy outcomes caused by early pregnancy infection with WH3 and WH6:The increase of the abortion rate,the intrauterine growth retardation and the bleeding of the fetal mice were observed in all mice infected with WH3 and WH6,and the adverse pregnancy outcome caused by the WH3 strain is more obvious.(2)The effect of WH3 and WH6 strains in early pregnancy on the decidual tr NK cell(CD49a~+Eomes~+tr NK)subgroups and their secretion of GPFs:early pregnancy infection leads to the proportion of CD49a~+Eomes~+tr NK cells decreased from 59.3±2.7%to 43.1±2.1%(WH3 group)and 48.6±1.4%(WH6 group),and the proportion of this cell subset in the second trimester was reduced from 41.0±3.2%to 8.6±2.9%(WH3 group)and 21.2±2.5%(WH6 group).The proportions of cell subsets secreting PTN,OPN and OGN in early pregnancy in nonifection group were 45.2±4.5%,51.1±2.1%,53.0±2.9%,respectively.While the proportions of in WH3 infection group were 22.7±2.2%,17.2±2.7%and 37.6±3.0%,respectively,and the proportions in WH6 infection group were 28.6±1.9%,28.4±1.9%,and 40.8±2.3%respectively;The proportions of cell subsets in the second trimester in nonifection group were 28.0±2.3%,35.9±1.4%,and 42.9±3.0%,and decreased to 17.3±3.5%,11.1±1.8%,and 17.4±3.8%,respectively,in WH3 infection group,while in WH6 infection group decreased to 24.8±2.1%,25.8±1.2%,28.0±2.8%,respectivly;As the infection progresses,the second trimester declines more than the early trimester,and the decline caused by the WH3 strain infection is more obvious.(3)Improvement of adverse pregnancy outcomes by supplemention of GPFs:The expression of Neu N in the cortex,the development of fetal rat bone tissues(including the spine and limbs),and the increase in the inner diameter of the spiral artery were found significantly improved after supplemention with PTN and OPN,while no significant improvement on the development of eye tissues after supplemention with GPFs.Conclusions:Mouse models infected with WH3 and WH6 in early pregnancy were successfully established.Both WH3 and WH6 can cause adverse pregnancy outcomes in mice.Infection of WH3 and WH6 in early pregnancy can significantly reduce the proportion of CD49a~+Eomes~+decidual tr NK and the secretion of GPFs(including PTN,OPN,OGN),thus affecting the remodeling of maternal spiral artery and the development of fetal nervous system and bone tissue thus affecting the remodeling of maternal spiral artery and the development of fetal nervous system and bone tissue. |
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