【摘要】 目的:探讨体外培养的人早孕绒毛膜滋养细胞在抗子宫内膜抗体(EmAb)环境中,低分子肝素对其增殖及凋亡能力的调节作用。方法:胰蛋白酶/DNA酶Ⅰ联合消化,通过Percoll细胞分离液纯化得到绒毛滋养细胞。按照EmAb(+)血清未加低分子肝素(A1组)、EmAb(+)血清加低分子肝素(A2组)、EmAb(-)血清未加低分子肝素(B1组)、EmAb(-)血清加低分子肝素(B2组)4个分组进行培养24h后,四甲基偶氮唑蓝比色法(MTT)检测滋养细胞细胞增殖情况,以490nm处的平均吸光度(A)值表示;采用免疫荧光化学法检测增殖细胞核抗原(PCNA)的表达水平,以平均荧光灰度值表示;采用流式细胞术检测细胞凋亡,以细胞凋亡率(%)表示。结果:(1)A1组细胞490nm处的A值为0.1007±0.0125,A2组为0.1982±0.0135,两者比较,差异有统计学意义(P<0.05);B1组细胞490 nm处的A值为0.2117±0.0106,B2组为0.2211±0.0080,两者比较,差异无统计学意义(P>0.05);(2)A1组细胞PCNA的表达水平为9.77±0.61,A2组为20.96±1.31,两者比较,差异有统计学意义(P<0.05);B1组细胞PCNA的表达水平为21.40±1.93,B2组为22.42±1.70,两者比较差异无统计学意义(P>0.05);(3)A1组细胞凋亡率为(2.64±0.10)%,A2组为(0.82±0.02)%,两者差异有统计学意义(P<0.05);B1组细胞凋亡率为(0.79±0.03)%,B2组为(0.76±0.03)%,两者差异无统计学意义(P>0.05)。结论:EmAb能抑制人早孕绒毛膜滋养细胞增殖,并促进其凋亡,这可能是导致自然流产的原因之一,而低分子肝素能够逆转这一过程。更多还原

【Abstract】 Objective:To investigate the modulation of low-molecular weight heparin on proliferation and apoptosis of cultured human cytotrophoblast cells under free EmAb environment.Methods:Chorionic villi tissue of normal 6～8 weeks pregnancy were obtained.Cytotrophoblast cells were collected by trypsin-DNase I digestion and Percoll gradient centrifugation.They were cultured under EmAb(+)serum group and EmAb(-)serum group which contained low-molecular weight heparin or not.After 24 hours,the 490 nm value,PCNA and apoptosis rate were detected by MTT,immunofluorescence and flow cytometry respectively.The proliferation and apoptosis of cultured human cytotrophoblast cells were evaluated.Results:In the culture systems with EmAb(+)serum,EmAb(+)serum and low-molecular weight heparin,EmAb(-)serum,EmAb(-)serum and low-molecular weight heparin,the 490 nm values were 0.1007±0.0125,0.1982±0.0135,0.2117±0.0106,0.2211±0.0080,the mean fluorescence grey scale values of PCNA were 9.77±0.61,20.96±1.31,21.40±1.93,22.42±1.70,the apoptosis rates were 2.64%±0.10%,0.82%±0.02%,0.79%±0.03%,0.76%±0.03%.Conclusion:EmAb can influence the prospective potency of the cytotrophoblast cells,inhabit cell proliferation and accelerate cell apoptosis,it will lead to spontaneous abortion,whereas low-molecular weight heparin can increase the ratio of pregnancy by reversing such effects in those patients.更多还原