The Effects Of Anticardiolipin Antibody Positive Blood Sera From Patients With Recurrent Spontaneous Abortion On The Development Of BeWo Cell And Modulation Of Aspirin

Objective Recurrent spontaneous abortion(RSA)is defined as two or more consecutive pregnancy or fetus(weight≤500g)losses prior to the 20th week of gestation,it affects up to 5%of fertile couples recorded by literatures.The etiology of recurrent spontaneous abortion is often multiplicate,with much controversy regarding diagnosis and treatment.Currently accepted etiologic causes include genetics,anatomical,endocrine,placental anomalies,hormonal problems,infection,smoking and alcohol consumption,exposure to environmental factors,psychological trauma and stressful life event,certain coagulation and immunoregulatory protein defects.Detection of an abnormality in any of these areas may result into specific therapeutic measures,with varying degrees of success.Howrever,about 50%cases of RSA remains unexplained. Reproductive immunology studies suggest that autoimmune factors may be the causes of RSA and some unexplained RSA cases is found to be associated with certain autoantibodies.Autoimmune factors represent the immunologic response of the mother to a pregnancy.Various autoantibodies that can cause abortion in women with RSA are antiphospholipid antibodies(APA),antinuclear antibodies (ANA),antithyroid antibodies(ATA)and so on,in which anticardiolipin antibodies(ACA)and Lupus anticoagulant(LC)are reported a higher prevalence. Thrombosis have been proposed the mechanism by which aPLA may cause pregnancy loss.Aspirin is generally considered to suffice in patients whose aPLA antibodies were positive because of its anticoagulant effect.Di Somone pointed out that besides thrombosis,the direct impairment of autoantibodies on trophoblast maybe also the cause of RSA.Researches about this aspect are uncommon.Using the detection of indexes of trophoblast development,this study is to evaluate the adverse effects of anticardiolipin antibody(ACA)positive blood sera from patients who suffer from recurrent spontaneous abortion on the development of trophoblast and determine the role of aspirin in regulating these functions.Methods Research objective:8 patients with recurrent pregnancy loss whose anticardiolipin antibody was positive were selected as research group and 8 healthy women with history of term delivery were selected as control group.2.Test methods2.1 Specimen collection Sera were collected from two groups in sterile condition and stored at -20℃.2.2 Culture of BeWo cell The culture systems were:ham F-12 culture media (free of fetal bovine serum)containing(1)sera of control group(2)sera of control group,aspirin(3)sera of research group(4)sera of research group,aspirin.The final serum concentration was 10%and for aspirin studies,25ug/ml aspirin were added.2.3 Evaluate the development of BeWo cell Viability was assessed by using MTT reduction test,secretion ofβ-HCG promoted by forskolin by ELISA and apoptosis applying flow cytofluorometry using M30 antibody.Results In the control group,the 490nm OD mean value was 0.2414±0.0320,β-HCG concentration was 22.9900±2.2568mIU/ml,M30 level was 0.7900±0.1786%;there were significant difference between the control group and the study group,the mean values of the latter group were 0.2061±0.0191,19.4488±1.8132 mIU/ml and 4.7868±0.5170%respectively,p＜0.05.Aspirin have no significant effect on the control group,the 490nm OD mean value was 0.2411±0.0320,β-HCG concentration was 23.2288±2.0581mIU/M1,M30 level was 0.6325±0.1701%,p＞0.05.Aspirin can attenuate ACA+ srea-induced adverse effects on BeWo cell in study group,the 490nm OD mean value was 0.2369±0.0311,β-HCG concentration was 21.5675±1.3338mIU/M1,M30 level was 2.7037±0.8920%,p＜0.05.Conclusion Besides thrombosis,ACA have direct effects on trophoblast biology. They can affect cell viability,induce apoptosis and have adverse effect on syneytiotrophoblast formation and hormone secretion.Aspirin can reverse these effects in some degree which providing an additional mechanism to explain the clinical benefits on recurrent pregnancy loss which is its direct modulation on trophoblast development.Significance The data presented in our study provide further evidence for using aspirin to treat RSA and meanwhile facilitate better treatment of placental dysfunction.Medicines regulating cell function may paly a role in the treatment of RSA.