Effect Of Drug-containing Serum Of Ramie Root Mixture On Trophoblast Proliferation In AMA With Threatened Abortion

Objective:The culture results of primary trophoblasts of AMA with threatened abortion of kidney deficiency and blood heat were compared with the serum containing low,middle and high doses of Ramie root mixture.the mechanism of Ramie root mixture in promoting trophoblast proliferation was discussed from the aspects of trophoblast growth characteristics,proliferation and apoptosis,changes of serum hormone levels,hormone receptors and immune tolerance mechanism.Methods:The solution of Ramie root mixture and dydrogesterone solution were prepared,and the rats were randomly divided into two groups and fed for 6 days.the drugcontaining serum of low,medium and high concentration Ramie root mixture group,dydrogesterone group and control group were prepared accordingly,and the complements were inactivated and sealed.The aseptic fresh curettage tissue was obtained,and some samples were taken for histopathological staining,and the results were observed and analyzed under microscope.Primary human trophoblasts were extracted from some tissues and cultured and detected with drug-containing serum.The growth status of trophoblasts was observed and photographed under periodic microscope,and the cell proliferation curve was drawn by cell counting with trypan blue.The secretion levels of E2,P.β-HCG,ER,PR.IL-2 and INF-γ of trophoblasts were detected by ELISA method after cultured with drug-containing serum for 72 hours.The effect of drug-containing serum of Ramie root mixture on the proliferation and apoptosis of human trophoblasts was detected by CCK-8 counting and flow cytometry.Results:1.After the primary human AMA trophoblast cells were extracted and cultured,the purity was more than 95%under the microscope.The primary cells were digested and counted with trypan blue staining solution,and the cell survival rate was 87%.2.Compared with the control culture group and the didrone culture group,the proliferation of trophoblasts in the Ramie root mixture culture group was faster,the growth and differentiation activity was stronger,the process of apoptosis was delayed,and the cell growth state was optimized and improved(p<0.05).3.The growth status of trophoblasts was observed.Compared with control culture group and dydrogesterone culture group,the growth status of trophoblasts in Ramie root mixture group was optimized,the biological characteristics and function of trophoblasts were enhanced,and there were significant proliferation changes.there was a certain growth cycle rule between proliferation and apoptosis,and the number of proliferation reached the peak and maintained stable growth at 72 hours.4.CCK-8 proliferation test showed that with the prolongation of the culture time of drug-containing serum,the cell absorbance and cell concentration increased.The viability of trophoblasts in Ramie root mixture culture group was higher than that in control culture group and dydrogesterone culture group(p<0.05).5.Flow apoptosis test showed that the total number of cells in Ramie root mixture culture group was more than that in control culture group and dydrogesterone culture group(p<0.05).The number of apoptotic cells in early and late stage was lower than that in control culture group and dydrogesterone culture group(p<0.05).6.ELISA detection of cell serum medium showed that the expression of E2,P,βHCG,ER and PR increased and the expression of IL-2 and INF-γ decreased significantly in Ramie root mixture culture group than that in control culture group and dydrogesterone culture group.Conclusions:Ramie root mixture plays an important role in the biological characteristics of trophoblasts,which can promote the proliferation of trophoblasts,enhance the ability of invasion and differentiation,and delay the process of apoptosis.At the same time,it can also increase the secretion levels of E2,P,β-HCG,ER and PR in serum(p<0.01).The increase of serum hormone secretion level and receptor can inhibit the secretion level of Thl-type(IL-2 and INF-y)to a certain extent(p<0.05),play a regulatory role in the Th1/Th2 immune tolerance mechanism,make the Th1/Th2 mechanism shift to Th2,participate in cellular immunity,and prevent the trophocells from being affected by adverse factors.In conclusion,the medicated serum of TCM can promote the proliferation of trophoblasts,delay cell apoptosis,and regulate the expression levels of serum hormones,hormone receptors and immune factors in an autocrine or paracrine way improve the maternal and fetal environment of advanced pregnancy in order to maintain pregnancy.