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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 effect of anti-cardiolipin antibody-positive serum on proliferation of human chorionic cell line BeWo in recurrent spontaneous abortion and the regulatory effect of heparin on the cell growth and development. Methods Thirty patients with recurrent spontaneous abortion with anticardiolipin antibody were selected as the study group and their serum was added to the culture system of BeWo cells containing heparin or heparin. Another 30 healthy women who had full-term labor history were selected as control Group, the same serum was added to heparin or heparin-free BeWo cell culture line, cultured for 24 h, the expression of PCNA in BeWo cells was detected by immunofluorescence staining, and the average fluorescence intensity Value, and the proliferation of BeWo cells was detected by MTT assay and expressed as average absorbance (A) value at 490 nm. Results The expression level of PCNA in the study group was 33.4 ± 2.2. The expression level of PCNA in the study group was 25.1 ± 2.3 without heparin, the difference was statistically significant (P <0.05). The serum level of PCNA The expression of PCNA in heparin group was 34.8 ± 3.1. The expression level of PCNA in control group without heparin was 33.8 ± 1.8. There was no significant difference between the two groups (P> 0.05). In the study group, the A value of serum added heparin at 0.054 ± 0.0027 was 0.0524 ± 0.0027, and the serum no heparin in study group was 0.0350 ± 0.0040, the difference was statistically significant (P <0.05) The value of A at 490 nm was 0.0560 ± 0.0033, while that of the control group without heparin was 0.0535 ± 0.0024. There was no significant difference between the two groups (P> 0.05). Conclusion Anticardiolipin antibody-positive serum in patients with recurrent spontaneous abortion may inhibit the proliferation of BeWo cells, which may be one of the causes of recurrent spontaneous abortion. However, heparin can reverse this effect.