Effects Of Autoantibody Positive Serum From Patients With Recurrent Pregnancy Loss On The Proliferation And Apoptosis Of BeWo Cells

Objective To investigate the effects of autoantibody positive serum from patients with recurrent pregnancy loss on the proliferation and apoptosis of BeWo cells as well as the mODulation of heparin on the growth of BeWo cell.MethODs 1.The culture of BeWo cell: the culture systems were : with heparin and normal serum(10%), normal serum(10%), heparin and serum with positive autoantibody(10%), serum with positive autoantibody(10%). 2. evaluate index:After 48 hours,the bcl-2 protain was detected by flow cytometry to evaluate the apoptosis of BeWo cell, and the PCNA and 490nm OD value were detected by immunofluorescence and MTT to evaluate the proliferation of BeWo cell.Results1. Evaluate of BeWo cell's proliferation1.1 MTT to evaluate the proliferation of BeWo cell.In the culture systems with heparin and normal serum, normal serum, heparin and serum with positive autoantibody, serum with positive autoantibody, the 490nm OD values were 0.054 4±0.006 8, 0.054 6±0.005 6, 0.047 8±0.003 0 and 0.032 1±0.005 8, respectively. There were significant difference between the the control group and autoantibODy positive group (p<0.01);There were significant differenc between the group of heparin and serum with positive autoantibody and the positive autoantibody group(p<0.01);there were no significant difference between the control group and the group of heparin and normal serum.1.2 The mean fluorecence gray scale values of PCNA(%)In the culture systems with heparin and normal serum, normal serum, heparin and serum with positive autoantibody, serum with positive autoantibody,the mean fluorecence gray scale values of PCNA were 35.11±1.25 35.84±1.04 35.39±1.31 and 26.14±3.27. There were significant difference between the the control group and autoantibody positive group (p<0.01);There were significant differenc between the group of heparin and serum with positive autoantibody and the positive autoantibody group(p<0.01);there were no significant difference between the control group and the group of heparin and normal serum.2. Evaluate of BeWo cell's apoptosisIn the culture systems with heparin and normal serum, normal serum, heparin and serum with positive autoantibody, serum with positive autoantibody, the expression of bcl-2 proteins were (34.26±1.22)%, (33.81±1.55)%, (32.81±1.44)% and (25.57±1.49)% , There were significant difference between the the control group and autoantibody positive group (p<0.01);There were significant differenc between the group of heparin and serum with positive autoantibody and the positive autoantibody group(p<0.01);there were no significant difference between the control group and the group of heparin and normal serum.Conclusion The serums with autoantibody positive can influence the prospective potency of BeWo cell by promoting cell apoptosis and inhibiting cell proliferation in vitro. Thus its direct influence on biological behaviour of the trophoblastic cell will lead to recurrent pregnancy loss, whereas heparin can increase the achievement ratio of pregnancy by reversing the effects on those patients.