Effects Of Bushen Antai Granule On VEGF And VEGFR2 In Placenta Vascular Cells Of RSA Mice

1 Objective To observe the effect of Bushen Antai Granule on embryo loss rate and VEGFR2 protein expression in placenta microvascular endothelial cells,vascular smooth muscle cells and fibroblasts of mice with recurrent abortion,and to explore the mechanism of Bushen Antai Granule on fetal protection at the molecular and cellular level,so as to provide a new theoretical basis for clinical application of Bushen Antai Granule on RSA.2 Methods RSA mice model was established by mating healthy female CBA / J mice with male DBA / 2 mice in 2:1 cage.The normal pregnant mice model was established by mating healthy female CBA / J mice with male BALB / c mice in 2:1 cage.Check the vagina of female rats from 5:00 to 6:00 in the morning every day.If there is a Yin suppository,the model is made successfully and it is recorded as the first day of pregnancy;if there is no Yin suppository,continue to close the cage.There were 50 mice with recurrent abortion and 10 normal pregnant mice.The successful RSA mice were randomly divided into five groups: model group,progesterone group,Chinese medicine high,medium and low dose group,10 mice in each group.From the first day of pregnancy,the normal pregnant group and model group were given distilled water gavage;the progesterone group was given 26 mg / kg / D progesterone capsule;the Bsat high,medium and low dose groups were given 35.1 g / kg / D,11.7 g / kg / D and 3.9 g/ kg / D distilled water dilution gavage,16:00-17:00 every day for 15 days.24 hours after the last administration,the mice were killed,and the complete uterus of each group was isolated.The placenta microvascular endothelial cells,vascular smooth muscle cells and fibroblasts were extracted and cultured under the body microscope.The embryo loss of each group of mice was observed and recorded by naked eyes.Western blot and immunofluorescence were used to detect the expression of VEGF and VEGFR2 in placenta microvascular cells,smooth muscle cells and fibroblasts of pregnant mice.3 Results3.1 embryo loss rate: compared with the normal group,the abortion rate of RSA mice in the model group was significantly increased(P < 0.01).Compared with the model group,the abortion rate of RSA mice in the western medicine group,the middle dose of Bushen Antai granules and the high dose of Bushen Antai granules decreased significantly(P < 0.01).3.2 Western blot was used to detect the expression of VEGF and VEGFR2 protein in placenta microvascular endothelial cells of pregnant mice in each group.Compared with the normal group,the expression of VEGF and VEGFR2 protein in the recurrent abortion model group decreased significantly(P < 0.01).Compared with the model group,the expression of VEGF and VEGFR2 protein in the western medicine group and Bushen Antai granule were significantly higher(P < 0.01).Compared with the high dose group,the expression of VEGF,VEGFR2 and VEGF protein in the middle and low dose group decreased significantly(P < 0.01),but the expression of VEGFR2 protein in the western medicine group did not decrease significantly(P > 0.05).3.3 Western blot was used to detect the expression of VEGF and VEGFR2 protein in the placental smooth muscle cells of the pregnant mice in the recurrent abortion model group compared with the normal group(P < 0.01).Compared with the model group,the expression of VEGF and VEGFR2 protein in the western medicine group and Bushen Antai granule were significantly higher(P < 0.01).Compared with the high-dose group,the expression of VEGF and VEGFR2 in the middle and low-dose group decreased significantly(P < 0.01);the expression of VEGF and VEGFR2 in the western medicine group did not decrease significantly(P > 0.05).3.4 Western blot was used to detect the expression of VEGF and VEGFR2 protein in the placenta fibroblasts of pregnant mice in each group.Compared with the normal group,the expression of VEGF and VEGFR2 protein in the recurrent abortion model group decreased significantly(P < 0.01).Compared with the model group,the expression of VEGF and VEGFR2 protein in the western medicine group and Bushen Antai granule were significantly higher(P < 0.01).Compared with the high-dose group,the expression of VEGF and VEGFR2 in the middle and low-dose group decreased significantly(P < 0.01);the expression of VEGF and VEGFR2 in the western medicine group did not decrease significantly(P > 0.05).3.5 the expression of VEGF and VEGFR2 protein in placenta microvascular endothelial cells of each group was significantly lower than that of the normal group(P< 0.01).Compared with the model group,the expression of VEGF and VEGFR2 protein in the western medicine group and Bushen Antai granule were significantly higher(P <0.01).Compared with the high dose group,the expression of VEGF,VEGFR2 in the middle and low dose group and VEGFR2 in the western medicine group decreased significantly(P < 0.01),while the expression of VEGF in the western medicine group did not decrease significantly(P > 0.05).3.6 the expression of VEGF and VEGFR2 protein in the placental smooth muscle cells of each group was significantly lower than that of the normal group(P < 0.01).Compared with the model group,the expression of VEGF and VEGFR2 protein in the western medicine group and Bushen Antai granule were significantly higher(P < 0.01).Compared with the high-dose group,the expression of VEGF and VEGFR2 in the middle and low-dose group decreased significantly(P < 0.01),while the expression of VEGF and VEGFR2 in the western medicine group did not decrease significantly(P >0.05).3.7 the expression of VEGF and VEGFR2 protein in the placenta fibroblasts of each group was significantly lower than that of the normal group(P < 0.01).Compared with the model group,the expression of VEGF and VEGFR2 protein in the western medicine group and Bushen Antai granule were significantly higher(P < 0.01).Compared with the high-dose group,the expression of VEGF and VEGFR2 in the middle and low-dose group decreased significantly(P < 0.01),while the expression of VEGF and VEGFR2 in the western medicine group did not decrease significantly(P >0.05).4 Conclusion Bushen Antai granule can activate the VEGF / VEGFR2 signal pathway,promote the angiogenesis,increase the number of capillaries,improve the formation of blood vessels at the maternal fetal interface,create a good vascular network environment,provide sufficient nutrition for the development of embryo,and thus reduce the expression of VEGF and VEGFR2 protein in microvascular endothelial cells,smooth muscle cells and fibroblasts The rate of embryo loss.