肝素对复发性流产患者抗心磷脂抗体阳性血清抑制人绒毛膜癌细胞增殖的调节作用

Effects of heparin and anticardiolipin antibodies positive serum from patients with recurrent pregnancy loss on proliferation of BeWo cells

目的探讨复发性流产患者抗心磷脂抗体阳性血清对人绒毛膜细胞株 BeWo 细胞增殖能力的影响,以及肝素对该细胞生长发育的调节作用。方法选择抗心磷脂抗体阳性的复发性流产患者30例为研究组,将其血清加入到含肝素或不含肝素的 BeWo 细胞培养系中;另选择有足月分娩史的健康妇女30例为对照组,同样将其血清加入到含肝素或不含肝素的 BeWo 细胞培养系中,均培养24 h 后,采用免疫荧光化学法检测 BeWo 细胞增殖细胞核抗原(PCNA)的表达水平,以平均荧光灰度值表示;四甲基偶氮唑蓝比色法(MTT)检测 BeWo 细胞增殖情况,以490 nm 处的平均吸光度(A)值表示。结果研究组血清加肝素者 PCNA 的表达水平为33.4±2.2、研究组血清未加肝素者 PCNA的表达水平为25.1±2.3,两者比较,差异有统计学意义(P<0.05);对照组血清加肝素者 PCNA 的表达水平为34.8±3.1、对照组血清未加肝素者 PCNA 的表达水平为33.8±1.8,两者比较,差异无统计学意义(P>0.05)。研究组血清加肝素者490 nm 处的 A 值为0.0524±0.0027,研究组血清未加肝素者为0.0350±0.0040,两者比较,差异有统计学意义(P<0.05);对照组血清加肝素者490 nm 处的 A值为0.0560±0.0033、对照组血清未加肝素者为0.0535±0.0024,两者比较,差异无统计学意义(P>0.05)。结论复发性流产患者抗心磷脂抗体阳性血清抑制 BeWo 细胞增殖,可能是导致复发性流产的原因之一;而肝素能够逆转这一作用。

Objective To investigate the effects of anticardiolipin antibodies positive serum from patients with recurrent pregnancy loss on the proliferation of BeWo cells as well as the modulation of heparin on the growth of BeWo cell. Methods Thirty patients with recurrent pregnancy loss whose anticardiolipin antibodies were positive and thirty healthy women with a history of term delivery were selected. Their sera were separately added to BeWo cell culture systems which contained either heparin or not. After 24 hours, the PCNA and 490 nm A value were detected by immunofluorescence and methyl thiazolyl tetrazolium （ MTT）, and the proliferation of BeWo cell was evaluated. Results In the culture systems with heparin and normal serum, normal serum, heparin and serum with positive autoantibody, serum with positive autoantibody, the mean fluorescence gray scale values of PCNA were 34. 8 ± 3.1,33. 8 ± 1.8,33.4 ± 2. 2 and 25.1 ± 2. 3, the 490 nm A values were 0. 0560 ± 0. 0033,0. 0535 ± 0. 0024,0. 0524 ± 0. 0027 and 0. 0350 ± 0. 0040, respectively. Conclusions The serum with anticardiolipin antibodies from patients with recurrent pregnancy loss can influence the prospective potency of BeWo cell, inhibit cell proliferation in vitro. Through this direct effect on biological behaviour of the trophoblastic cell, it will lead to recurrent pregnancy loss, whereas heparin can increase the ratio of pregnancy by reversing such effects in those patients.