The Role And Clinical Significance Of GGT And Beclin-1 In Missed Abortion

Objective:Missed abortion(Missed abortion,MA)is a special type of abortion.With the liberalization of the two-child policy,the incidence of missed abortion is gradually increasing.At present,the pathogenesis of missed abortion is not very clear,and there is a lack of early prediction target.Some studies have suggested that oxidative stress and apoptosis may be involved in the occurrence of missed abortion.This study screened the clinical common blood indexes involved in oxidative stress,apoptosis and other metabolic pathways: ?-glutamyltransferase(gamma glutamyltransferase,GGT),lactic dehydrogenase(lactate dehydrogenase,LDH),adenosine deaminase(adenosine deaminase,ADA)and fibrinogen(Fibrinogen,FIB),and analyze its expression in peripheral blood of patients with missed abortion and normal pregnancy(Normal pregnancy,NP),and analyze the diagnostic value of each index.At the same time,the expression of autophagy key factors Beclin-1 and LC3 in pregnancy tissues(villi and decidua)of normal pregnancy and missed abortion patients were detected to explore the relationship between autophagy and the pathogenesis of missed abortion.Through the bioinformatics analysis of Beclin-1,the possible pathogenic mechanism was explored.Methods:1.The expression of GGT,LDH,ADA and FIB in 795 patients with missed abortion and 694 normal pregnancy patients from March 2014 to March 2017 were analyzed retrospectively.according to the average age,they were divided into age group 1(17-27 years old),age group 2(28-38 years old)and age group 3(39-49 years old).The expression differences of each age group were counted in order to understand the relationship between the age factors and the expression differences of each index.The hematological indexes were analyzed by ROC curve.2.From January 2017 to March 2017,36 patients with missed abortion were selected as missed abortion group,and 36 normal pregnant women who voluntarily terminated pregnancy were selected as normal pregnancy group.HE staining wasused to observe the characteristics of villi and decidua,immunohistochemistry was used to observe the localization of Beclin-1 in villi and decidua,RT-PCR was used to detect the expression of Beclin-1 mRNA in villi and decidua,and Western Blot was used to detect the expression of Beclin-1 protein and LC3II/LC3 I ratio in villi and decidua.3.Using STRING online software to draw the Beclin-1 protein interaction network diagram,and then using DAVID online analysis software to analyze the signal pathway enrichment of the screened interacting proteins,in order to predict the mechanism of Beclin-1 participation and the possible regulation of downstream proteins.Results:1.Compared with normal pregnancy patients,the levels of serum GGT and ADA were significantly increased in patients with missed abortion(GGT : MA :17.68�9.85 IU / L;NP:14.57�7.93 IU / L,P <0.0001;ADA:MA:6.05�3.94 IU /L;NP:5.67�3.36 IU / L,P<0.05).The level of plasma FIB decreased in patients with missed abortion(MA:4.31�0.91 g / L;NP:4.44�0.96 g / L,P<0.01).There was no significant difference in serum LDH level between the two groups(MA :148.05�30.46 IU / L;NP:149.34�27.88 IU / L,P>0.05).In further age groups,it was found that the expression of ADA and FIB was only statistically different in the age group of 17 to 27 years old,and there was significant difference in GGT among all age groups.2.The area under the ROC curve of GGT,ADA,FIB and LDH was 0.643?0.548?0.539 and 0.527 respectively.3.HE staining showed that compared with the villi and decidua of normal pregnancy,the syncytiotrophoblast and cytotrophoblast of missed abortion villi were significantly atrophied and thinned,nuclear pyknosis appeared in trophoblast,villi tissue showed different degrees of degeneration,villous stroma showed fibroid degeneration.The number of blood vessels in decidual tissue of patients with missed abortion was relatively reduced,and the vascular cavity was irregular,which showed loose tissue structure,swelling of glandular epithelial cells,unclear display ofglandular cavity,increased volume of decidual cells,shallow staining and obvious edema.4.Immunohistochemical staining showed that the expression pattern of Beclin-1protein was up-regulated in villi and decidua of missed abortion.5.Compared with normal pregnancy group,Beclin-1 mRNA was up-regulated in missed abortion villi(NP:1.00�0.17;MA:1.48�0.24,P<0.001)and decidual tissue(NP:1.00�0.37;MA:2.30�0.37,P<0.001).Beclin-1 protein was up-regulated in missed abortion villi(NP:1.00�0.34;MA:2.90�0.60,P<0.001)and decidual tissue(NP:1.00�0.29;MA:3.00�0.73,P<0.001).The difference was statistically significant.LC3II/LC3 I was up-regulated in missed abortion villi(NP:1.00�0.27;MA:2.34�0.46,P<0.001)and decidual tissue(NP:1.00�0.02;MA:1.89�0.33,P<0.001).The difference was statistically significant.6.30 proteins interacting with Beclin-1 were screened by STRING,among which the downstream proteins ATG14 and PIK3C3 play an important role.Signal pathway enrichment analysis showed that Beclin-1 and its interacting proteins were significantly enriched in autophagy,apoptosis,cancer,inflammation and other pathways.Conclusion:1.Compared with normal pregnant women,the levels of GGT and ADA in peripheral blood of patients with missed abortion were up-regulated,and the levels of FIB were down-regulated,and there were statistical differences.After further age groups,it was found that the expression of serum ADA and FIB was only significantly different in the age group of 17 to 27 years old,and there was significant difference in GGT among all age groups.At the same time,the area under the ROC curve of GGT is the largest and the diagnostic value is relatively high,so GGT is expected to become a reliable target for predicting missed abortion.2.The expression of Beclin-1 mRNA and protein and the ratio of LC3II/LC3 I were up-regulated in villi and decidua of patients with missed abortion,suggesting that autophagy was involved in the occurrence of missed abortion.3.ATG14 and PIK3C3 may be downstream proteins closely related to Beclin-1.The network system formed by the interaction with other related proteins may act on autophagy,apoptosis,tumor,inflammation and other pathways,which provides a direction for further study of the mechanism of Beclin-1 in missed abortion.