| Objective: With the VEGF signaling path as the breakthrough, concerning Bushenantai Granule, the research is to observe the condition of the uterine and embryonic development of the mouse model of recurrent spontaneous abortion and the embryonic loss rate, and to detect the protein expression of VEGF, VEGFR2, RAS and MAPK in the decidual tissue, so as to figure out the mechanism of Chinese herbs that are used to reinforce the kidney and invigorate the spleen, in the aspect of pregnancy vascular remoulding when treating RSA. As a result, it is supposed to provide a theoretical basis for Bushenantai Granule in the treatment of RSA.Methods :The mouse model of recurrent spontaneous abortion was established by female mice CBA/J and male mice DBA/2, the mouse model of normal pregnancy established by female mice CBA/J and male mice BALB/c, with the female and the male mated by the ratio of 2:1. After one week of adaptive feed, 60 pregnant mice as models were selected, all of which divided into six groups: the normal pregnancy group,the pregnancy model group of recurrent spontaneous abortion, the western medicine group, the Chinese medicine group of low dose, the Chinese medicine group of medium dose, the Chinese medicine group of high dose. The gavage for mice started from the first day of pregnancy: the pregnancy model group of recurrent spontaneous abortion,i.e., the model group, were gavaged with diluted distilled water; the western medicine group were given progesterone capsules with the dose of 0.4mg/kg.d; the Chinese medicine group of low dose, i.e., the group of low-dose Bushenantai Granule, would be administered with Bushenantai Granule, the dose of which was 3.9g/kg.d; the Chinese medicine group of medium dose, i.e., the group of medium-dose Bushenantai Granule,would be administered with Bushenantai Granule, the dose of which was11.7g/kg.d; the Chinese medicine group of high dose, i.e., the group of high-dose Bushenantai Granule,would be administered with Bushenantai Granule, the dose of which was 35.1g/kg.d.The successive administration was given 15 days. The mice were executed in 24 h after the last administration, whose uteri were then pulled out. The next step was to observe the uterine development after dissection, calculate the embryonic loss rate, and detect the protein expression of VEGF, VEGFR2, RAS and MAPK in the decidual tissue.Results:1. The observation finds that the uteri and embryos in the normal pregnancy group are well-developed, being rufous, bead-like shaped, and in a relatively uniform size.Besides, there are almost no notable points of blood stasis in the uteri, neither evident bleeding in the implantation sites. The development of the uteri and embryos in the model group are poor, being dark brown, shaped in a bamboo spine, and in a poor uniformity. There are evident blood stasis in the uteri, and the embryonic tissues have disappeared or partly disappeared in the implantation sites. As for the western medicine group and all the low-dose, medium-dose, and high-dose Bushenantai Granule groups,uteri and embryos are well-distributed, being rufous, with no evident bleeding inside.The embryonic development is good.2. Compared to the normal pregnancy group, the embryonic loss rate in the control model group, rose significantly(P<0.05). When Comparing with the pregnancy model group of recurrent spontaneous abortion, the western medicine group, the group of medium-dose Bushenantai Granule, the group of high-dose Bushenantai Granule, the rate rose significantly, P<0.05, with statistical difference, while the group of low-dose Bushenantai Granule declined sharply, P>0.05, with the difference not statistically significant.3. Compared to the normal pregnancy group, the VEGF expression, P<0.01, in the pregnancy model group of recurrent spontaneous abortion dropped sharply. However,when compared to the pregnancy model group of recurrent spontaneous abortion, the VEGF expression, P<0.01, in the western medicine group and all the low-dose,medium-dose, and high-dose Chinese medicine groups, rose evidently; while compared to the western medicine group and the Chinese medicine ones of low dose and medium dose, the expression level of VEGF, P>0.05, in the high dose group rose significantly.4. Compared to the normal pregnancy group, the VEGFR2 expression, P<0.01, in the pregnancy model group of recurrent spontaneous abortion, dropped sharply. when compared to the pregnancy model group of recurrent spontaneous abortion, the VEGFR2 expression, P<0.01, in the western medicine group and all the low-dose,medium-dose, and high-dose Chinese medicine groups, rose evidently; while compared to the western medicine group and the Chinese medicine ones of low dose and medium dose, the expression level of VEGFR2, P>0.05, in the high dose group, rose significantly.5. Compared to the normal pregnancy group, the RAS expression, P<0.01, in the pregnancy model group of recurrent spontaneous abortion, dropped sharply. when compared to the pregnancy model group of recurrent spontaneous abortion, the RAS expression, P<0.01, in the western medicine group and all the low-dose, medium-dose,and high-dose Chinese medicine groups, rose evidently; while compared to the western medicine group and the Chinese medicine ones of low dose and medium dose, the expression level of RAS, P>0.05, in the high dose group, rose significantly.6. Compared to the normal pregnancy group, the MAPK expression, P<0.01, in the pregnancy model group of recurrent spontaneous abortion dropped sharply. However,when compared to the pregnancy model group of recurrent spontaneous abortion, the MAPK expression, P<0.01, in the western medicine group and all the low-dose,medium-dose, and high-dose Chinese medicine groups, rose evidently; while compared to the western medicine group and the Chinese medicine ones of low dose and medium dose, the expression level of MAPK, P>0.05, in the high dose group rose significantly.Conclusion:1. Concerning mice of recurrent spontaneous abortion, Bushenantai Granule may have a positive effect on the uterine and embryonic development.2. The embryonic loss rate of the mouse model of recurrent spontaneous abortion can be sharply reduced by Bushenantai Granule, which has the efficacy of preventing miscarriages.3. It is indicated by the results of western blot of VEGF, VEGFR2, RAS and MAPK,that the mechanism of Bushenantai Granule's efficacy for the recurrent spontaneous abortion might promote the expression level of RAS and MAPK in the VEGF signaling path's downstream, which, meanwhile, is also true of that of VEGF and VEGFR2 in the decidual tissue of mice of recurrent spontaneous abortion, better the vascularization of the maternal-fetal interface of the mouse model of recurrent spontaneous abortion, and maintain pregnancy, thus reducing the embryonic loss rate of mice of recurrent spontaneous abortion. |
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