构建并利用绒毛外滋养细胞组织芯片检测磷酸化信号转导和转录活化因子3在子痫前期患者的表达

Detection of phosphorylated signal transducer and activator of transcription 3 in extravillous trophoblast cells of preeclampsia by placental tissue microarray

目的构建子痫前期患者绒毛外滋养细胞（extravillous trophoblast, EVCT）组织芯片，并利用该组织芯片检测磷酸化信号转导和转录活化因子3 （phosphorylated signal transducer and activator of transcription 3,pStat3）在子痫前期EVCT中的表达水平，探讨Stat3信号途径在子痫前期病理过程中的作用。方法收集2007年12月12日至2010年12月31日郑州大学第三附属医院80例子痫前期和58例正常妊娠孕妇的胎盘组织用来构建组织芯片，利用波形蛋白、细胞角蛋白和人类白细胞抗原-G抗体进行免疫组织化学染色验证EVCT组织芯片，采用免疫组织化学半定量方法检测pStat3在子痫前期和正常妊娠孕妇胎盘EVCT组织的表达情况。采用非参数秩和检验、Kruskai-wallisH检验、t检验和卡方检验进行统计分析。结果57例子痫前期（109个组织点）和31例正常妊娠孕妇的胎盘块（65个组织点）适于EVCT组织芯片的构建。组织芯片中EVCT细胞角蛋白染色阳性，波形蛋白染色阴性，人类白细胞抗原-G染色阳性，符合EVCT的特点，86．4％（76／88）的样本保留了EVCT，组织芯片构建成功。子痫前期患者胎盘EVCT中pStat3阳性率为51．1％（24／47），重度子痫前期组阳性率为52．3％（23／44），早发型子痫前期组阳性率为50．0％（19／38），均显著低于正常妊娠组的72．4％（21／29），差异均有统计学意义（U分别为492．00、473．00和401．00，P均〈0．05）。结论EVCT组织芯片构建成功，利用该芯片可高通量检测pStat3的表达情况。pStat3信号转导途径可能参与了早发型和重度子痫前期的发生发展。

Objective To construct extravillous trophoblast （EVCT） tissue microarray and detect the expression of phosphorylated signal transducer and activator of transcription 3 （pStat3） in EVCT and to explore the role of Stat3 signal transduction pathway in the pathogenesis of preeclampsia. Methods Placentas of 80 pregnant women with preeclampsia and 58 normal pregnant women hospitalized in the Third Affiliated Hospital of Zhengzhou University from December 12,2007 to December 31,2010 were recruited for constructing EVCT tissue microarray. Vimentin,cytokeratin and human leukocyte antigen-G were used to verify EVCT tissue microarray immunohistochemically. The difference of pStat3 expression was detected between preeclampsia patients and normal pregnant women by immunohistochemical staining. Rank-sum test, Kruskai-Wallis H test, t-test and Chi- square test were used for statistical analysis. Results Placental tissues from 57 preeclampsia patients （109 tissue cores） and 31 normal pregnant women （65 tissue cores） were suitable for constructing EVCT tissue microarray. The target tissue was positive for both cytokeratin and human leukocyte antigen-G staining and negative for vimentin, which was in accordance with the characters ofEVCT tissue. Totally 86.4%（76/88） samples retained the target EVCT tissues, which meant EVCT tissue microarray was constructed successfully. The expression of pStat3 was significantly decreased in EVCT of preeclampsia patients （51.1%, 24/47）, the early onset （50.0%, 19/38） and severe preeclampsia patients（52, 3%,23/44） as compared to normal pregnant women （72.4% ,21/29） （U= 492.00, 473.00 and 401.00,P〈0.05 respectively）. Conclusions EVCT tissue microarray has been successfully constructed, and could be used to detect pStat3 expression, pStat3 signal transduction pathway may be involved in the development of preeclampsia.