| Follistatin is extracted from porcine follicular fluid single-chain glycoprotein,it has the effect of inhibiting the secretion of anterior pituitary gland cells of follicle stimulating hormone(FSH),and binding to activin FST is widely distributed in adrenal,pituitary,pancreas,uterine,muscle and other tissues,by binding to activin and neutralize its biological function,which jointly regulate the body’s life activities.Studies have shown that,FST is a germ regulating reproductive factors,maintain physiological pregnancy,fetal growth and development.Studies have confirmed that,follistatin expression is upregulated in women with decidual tissue during early pregnancy,and the expression of follistatin in the endometrium of normal non-pregnant women is reduced,but the relationship between the level of FST expression and the development of early decidua and embryonic death due to dysplasia development is still lacking in-depth research.This study explored the relationship between FST expression and early decidual development and spontaneous abortion by establishing a mouse pregnancy model.The relationship between the expression of FST and Activin A in decidua tissue and and the early embryonic death in early pregnancy was analyzed by detecting the expression of FST and Activin A in decidual tissue of early pregnant women and embryonic death patients.1.Methods(1)Serum FST and Activin A levels were detected: ELISA was used to detect serum FST and Activin A levels in normal pregnancy and embryonic death patients.(2)Detection of FST and Activin A expression in decidual tissue:RT-PCR and immunohistochemical staining were used to detect the expression of FST and Activin A m RNA and protein in decidual tissue of normal pregnancy and embryonic death patients.(3)Differential expression analysis of mRNA: gene expression profiling and RT-PCR were used to detect the mRNA expression levels of decidual tissue in different stages of pregnancy.(4)Detection of peripheral blood immune cells: the ratio of NK,T cells in uterus and peripheral blood of normal pregnant mice and pregnant mice on day 7 was measured by flow cytometry.2.Results(1)Changes in FST expression in mouse decidual tissue: the results of mRNA expression profiling showed that FST increased its expression in decidual tissue by more than 20 times,and its family member FSTL1 also increased more than 4 times.The decidua tissues of E5,E7 and E8 mice were further selected,and the expression of FST mRNA and Activin βA mRNA were detected by RT-PCR.The results showed that the expression of FST and Activin βA mRNA in the decidual tissue of pregnant mice was higher than that of normal unpregnant mice,and the expression of E7 mice was higher than that of E5 mice.(2)Changes in FST expression in decidual tissue of aborted mice: a female abortion model was established using female CBA/J mice: male DBA /2 mice.The expressions of FST and Activin βA mRNA in normal uterine decidua and aborted embryonic decidual tissue were detected by RT-PCR.The results showed that the expression of FST in normal uterine decidua was lower than that in aborted uterine decidual tissue.The expression level of Activin βA did not change significantly.(3)Changes of FST and Activin A levels in peripheral blood of patients with embryonic death: The results of ELISA showed that the serum FST level of embryonic death patients was significantly lower than that of normal pregnant women,while the level of Activin A did not change significantly between the two.(4)Expression of FST and Activin A in uterine decidua tissue of patients with embryonic death:Firstly,the expression of FST and Activin βA mRNA was detected by RT-PCR.The results showed that the expression level of FST mRNA in artificial abortion uterine decidua was lower than that in normal pregnant group.The expression level of Activin βA mRNA dign,t change significantly compared with normal early pregnancy group.Secondly,the expression of FST and Activin A protein was detected by immunohistochemical staining.The results showed that the expression of FST protein in decidual tissue of embryonic abortion patients wassignificantly lower than that of normal decidua in early pregnancy,and Activin A protein expression was no significant change,and this result was consistent with the results of the mRNA study.(5)Changes in immune cells during decidualization of the uterusPregnancy uterine decidualization not only has differences in molecular expression,but also the composition of decidual immune cells.Therefore,the study also used flow cytometry to analyze the changes of NK and T cells in decidual tissue.The results showed that the proportion of NK cells in the uterus of the 7th day of pregnancy was significantly higher than that of the normal non-pregnant mice,while the proportion of NK cells in the peripheral blood of the mice on the 7th day of pregnancy was lower than that of the normal non-pregnant mice.On the 7th day of pregnancy,the ratio of CD3+CD8+ T cells and CD3+CD4+ T cells in the uterus of the mice was not significantly different from that in the normal uterus of the pregnant mice.3.Conclusions Based on the above findings,the expression of FST increased during decidualization of the uterus during pregnancy,while the expression of decidual tissue decreased in embryonic death patients,while the expression of Activin A in the decidual tissue of embryonic death patients was not significantly different from that of normal pregnant decidua.This result suggests that FST expression is associated with decidualization of the uterus and early embryonic death.The study also showed that the proportion of NK cells in decidual tissue of mice was more obvious than that of T cells during decidualization of pregnancy.Therefore,not only the expression level of FST can be used as an important marker of decidualization,NK cell changes,but also an important marker of decidualization of the uterus.Therefore,detecting the expression level of FST and the proportion of NK cells can help clinically determine the early development of decidua,and it is helpful to prevent the occurrence of spontaneous abortion. |
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