The Expression Of RAS,MAPK,PI3K,AKT In Decidual Tissue Of Patients With RSA And Intervention Of Bushen Antai Granules On Mice With Abortion

1 Objective To observe the expression of Ras,MAPK,PI3K and AKT proteins downstream of VEGF angiogenesis signaling pathway in retinal abortion mice and decidual tissue of patients with recurrent spontaneous abortion,and to explore the effect of kidney-repairing by interfering with recurrent abortion mice.The effect of granules on its expression,and the molecular and biological mechanism of tonifying kidney and strengthening spleen Chinese medicine to promote angiogenesis at maternal-fetal interface,provide a theoretical basis for the application of Chinese herbal medicine for invigorating kidney and strengthening spleen.2 Methods Clinically,30 cases of patients with recurrent spontaneous abortion of renal deficiency who were admitted to the gynaecological clinic of the First Affiliated Hospital of Anhui University of Traditional Chinese Medicine in 2018 were used as observation groups.In the same period of normal early pregnancy,30 patients with induced abortion were used as the control group.The decidual tissue was obtained.The morphological changes of decidual tissue and the immunohistochemical SP method were used to detect the expression of Ras,MAPK,PI3K and AKT in the decidual tissue of the two groups.The results were statistically analyzed.analysis.50 mice with DBA/2×CBA/J Clark classic recurrent abortion model were randomly divided into RSA model group,western medicine control group,Chinese medicine high,medium and low dose groups,10 rats in each group,and BALB./c×CBA/J 10 normal pregnant mice,from the first day of pregnancy,were given different intervention methods for 15 days,24 hours after the last administration,the mice were sacrificed,the uterus was removed,and the embryos of 6 groups were observed.In the same situation,the decidua was exfoliated,and the decidual tissue morphology of each group was compared by HE staining.The expression of Ras,MAPK,PI3K and AKT protein in decidual tissue of 6 groups of mice were detected by immunohistochemistry and Western-blot technique.The situation is tested and analyzed,and the test data is statistically analyzed.3 Results Clinical study results 3.1 Observing the morphological changes of decidua in HE staining group Compared with the control group,the number of decidual tissue cells in the observation group was significantly reduced,the arrangement was disordered,the size was extremely irregular,some cells were squeezed and deformed,the gland atrophied,the nucleus became smaller,and some were fusiform,and the staining was obvious.Deepened,it is condensed,and even the nucleus disappears.The cytoplasm is edema,vacuolization changes,the interstitial cells are cellulose-like,the number of blood vessels is reduced,and the blood vessel wall is visible.3.2 Comparison of immunohistochemical method to determine the expression of decidual tissue-associated protein in two groups of patients 3.2.1 Expression of Ras and MAPK in decidua tissue of two groups of patients Ras and MAPK proteins were mainly expressed in the cytoplasm of glandular epithelial cells and vascular endothelial cells of the two groups of patients.Compared with the control group,the expression of Ras and MAPK protein in the decidual tissue of the observation group was decreased,and the difference was statistically significant(P<0.05).3.2.2 Expression of PI3K and AKT in decidual tissue of two groups of patients The PI3K and AKT proteins are mainly expressed in the cytoplasm of glandular epithelial cells and vascular endothelial cells in the two groups of patients,and a small amount is also expressed on the cell membrane.The expression of related proteins in the observation group was lower than that in the control group,and the difference was statistically significant(P<0.05).Experimental research results3.1 Comparison of embryo loss rate in each group of mice Compared with the normal group,the embryo loss rate of the model group was significantly increased(P<0.01).Compared with the model group,the embryonic loss rate of the western medicine group and the Chinese medicine high and medium dose group was significantly lower,and the difference was statistically significant(P<0.01).The embryo loss rate of the low dose group of Chinese medicine was lower than that of the model group.Academic significance(P<0.05).Compared with the western medicine group,the high-dose group of Chinese medicine had better curative effect,and the embryo loss rate of RSA mice was close to it(P>0.05).Compared with the high-dose group of traditional Chinese medicine,the embryonic loss rate of the middle and low dose groups of Chinese medicine was significantly lower(P<0.01).3.2 Comparison of morphological changes of decidua in each group of HE staining mice Compared with the normal group,the decidual tissue of the model group was degenerated,the number of decidual cells was significantly reduced,the cell arrangement was disordered,the cytoplasmic edema was obvious,the vacuole was changed,part of the nucleus disappeared,the nucleus was pyknosis,the staining was deepened,and the number of blood vessels was significantly reduced..Compared with the model group,the number of cells and the number of blood vessels in the decidua tissue of the western medicine group and the traditional Chinese medicine group increased,and there was no significant difference in the decidual tissue morphology between the western medicine group and the high-dose group.With the increase of the dose,the number of decidual cells and the number of blood vessels gradually increased,the cytoplasm gradually became rich,the cytoplasmic edema was relieved,the nucleolus gradually became lightly stained,became larger and more obvious,the wall became more intact,and the vascular endothelial cells were arranged.neat.3.3 Analysis of the expression of decidual tissue related proteins in each group of mice by immunohistochemistry 3.3.1 Immunohistochemical method was used to determine the expression of Ras and MAPK protein in decidua of mice in each group Ras and MAPK proteins were mainly distributed in the cytoplasm of mouse glandular epithelial cells and vascular endothelial cells,and no obvious expression was observed on the nucleus and membrane.Compared with the normal group,the expression levels of Ras and MAPK protein in the model group were significantly decreased(P<0.01).Compared with the model group,the expression of Ras and MAPK protein in the high,middle and low dose groups of western medicine group and Chinese medicine was significantly increased(P<0.01).Compared with the high-dose group of traditional Chinese medicine,the expression of Ras and MAPK protein in the middle and low dose groups of Chinese medicine decreased significantly(P<0.01).The expression of Ras and MAPK protein in western medicine group decreased,the difference was statistically significant(P<0.05).3.3.2 Immunohistochemical method was used to determine the expression of PI3K and AKT protein in decidua of mice in each group PI3K and AKT proteins are mainly distributed in the cytoplasm of mouse glandular epithelial cells and vascular endothelial cells,and a small amount of expression is also observed on the cell membrane.Compared with the normal group,the expression of PI3K and AKT protein in the model group was significantly decreased(P<0.01).Compared with the model group,the expression levels of PI3K and AKT protein in the high,middle and low dose groups of western medicine group and Chinese medicine group were significant.Elevated(P < 0.01).Compared with the western medicine group,there was no significant difference in the expression of PI3K and AKT protein in the high dose group(P>0.05).Compared with the high-dose group of traditional Chinese medicine,the expression levels of PI3K and AKT protein in the middle and low dose groups of Chinese medicine were significantly lower(P<0.01).3.4 Comparison of the expression of decidual tissue-associated protein in each group of mice by Western blot 3.4.1 Comparison of Ras and MAPK protein expression in decidual tissue of each group by Western blotCompared with the normal group,the expression levels of Ras and MAPK protein in the model group were significantly decreased(P<0.01).Compared with the model group,the expression levels of Ras and MAPK protein in the high,middle and low dose groups of western medicine group and Chinese medicine group were significantly increased(P<0.01).Compared with the high-dose group of traditional Chinese medicine,the expression of Ras and MAPK protein in the middle and low dose groups of Chinese medicine decreased significantly(P<0.01).The expression of Ras and MAPK protein in western medicine group decreased,the difference was statistically significant(P<0.05).3.4.2 Comparison of the expression of PI3K and AKT protein in decidual tissue of each group by Western blot Compared with the normal group,the expression of decidual-associated PI3K and AKT in the model group was significantly decreased(P<0.01).Compared with the model group,the western medicine group,the high-dose group of Chinese medicine,and the traditional Chinese medicine.The expression of PI3K and AKT in the decidua of the mice in the low dose group was significantly increased(P<0.01).Compared with the western medicine group,there was no significant difference in the expression of PI3K and AKT in the high dose group(P>0.05).Compared with the high-dose group of traditional Chinese medicine,the expression of PI3K and AKT in the decidual tissue of the middle and low dose groups of Chinese medicine was significantly lower(P<0.01).4 Conclusion 4.1 RSA patients and mice morphological changes in the morphology of the decidua,and maternal fetal vascular dysplasia,poor blood supply.Decidual angiogenesis is associated with the expression of Ras,MAPK,PI3K,and AKT-related proteins.4.2 Bushen Antai Granules with Bushen Jianpi Recipe can significantly improve the decidual tissue morphology of RSA mice,promote the regeneration of decidual blood vessels,and reduce the embryonic loss rate of RSA mice,so as to protect pregnancy.4.3 Possible Mechanisms of Bushen Antai Granules to Protect Pregnancy:(1)Bushen Antai Granules up-regulated the expression of Ras,MAPK,PI3K and AKT proteins in decidual tissue of RSA mice,and the protein expression level increased with the increase of the dose.Thereby promoting decidual angiogenesis,improving the blood supply of the decidual tissue,ensuring the smooth flow of blood at the maternal-fetal interface and the supply of embryonic nutrients,thereby reducing the embryo loss rate.(2)Bushen Antai Granules can promote the signal transduction of VEGF/Ras/MAPK and VEGF/PI3K/AKT pathways by improving the expression levels of Ras,MAPK,PI3K and AKT proteins in decidual blood vessels,and ensure the coordination of various signal molecules in the pathway.Thereby improving decidual angiogenesis and promoting embryonic development in RSA mice.