妊娠高血压综合征患者胎盘滋养细胞凋亡基因表达谱的研究

目的　采用基因芯片技术筛查妊娠高血压综合征 (妊高征 )患者胎盘滋养细胞凋亡基因的差异表达 ,探讨细胞凋亡与妊高征发病的关系。方法　妊高征患者 2 0例为妊高征组 ,正常孕妇 10例为对照组。采用脱氧核糖核苷酸末端转移酶介导的缺口末端标记 (TUNEL)方法检测两组孕妇胎盘组织滋养细胞凋亡情况。同时采用上海博星公司生产的基因芯片进行凋亡基因表达谱的筛查。判断基因差异表达的标准 :(1)cy5 (妊高征组 ) /cy3(对照组 )的自然对数绝对值 >0 6 9,cy3与cy5的信号相差 2倍以上。 (2 )cy3或cy5、cy3和cy5的信号值其中之一必须 >80 0。 结果　 (1)TUNEL检测显示 ,每 10 0 0 0 μm2 绒毛面积中滋养细胞凋亡数 ,妊高征组为 1 5 84个 ,对照组为 0 0 32个 ,两组比较 ,差异有极显著性 (P <0 0 1)。 (2 )基因芯片筛查有差异表达的凋亡基因 10个 ,占基因芯片筛查凋亡基因总数的 5 %。妊高征组胎盘组织中的凋亡基因表达均明显下调 ,即cy5 /cy3比值 <1。其中包括具有明显抗凋亡作用的SFRP2 、IAP2 、DHCY2 4及ATPIA1基因。结论　妊高征患者胎盘滋养细胞存在显著的凋亡现象 ,具有抗凋亡作用的基因表达明显下调 ,将可能导致胎盘滋养细胞凋亡 ,从而导致妊高征发病。

Apoptosis gene expression profiling of placental trophoblast cells in patients with pregnancy induced hypertension

OBJECTIVE: To explore the function of placental trophoblast cell apoptosis on the pathogenetic mechanism of pregnancy induced hypertension (PIH). METHODS: Apoptosis of trophoblast cells in 20 cases of PIH (PIH group) and in 10 cases of normal pregnancy (control group) were directly observed using the terminal-deoxynucleotidyl transferase mediated d-UTP nick end labeling (TUNEL) method. Apoptosis gene expression patterns were screened with gene chip provided by Poxing Company, Shanghai. Standards for differently expressed genes were: (1) An absolute value of the natural logarithm of cy5 (PIH group)/cy3 (control group) greater than 0.69 with a difference of signal of cy5 2 times over that of cy3. (2) The signal value either cy3 or cy5 must be greater than 800. RESULTS: (1) TUNEL test showed that the number of trophoblast cells apoptosis per ten thousand micro m(2) was 1.584 in the PIH group and 0.032 in the control group with significant difference between the two groups (P < 0.01). (2) Ten differently expressed apoptosis genes were obtained through gene chip test (occupy 5% of total apoptosis gene in the gene chip). There was a significant decrease of apoptosis gene expression in all of PIH patient placental tissues (i.e a ratio of cy5/cy3 less than 1). Among them, there were genes that possess significant anti-apoptosis functions (including SFRP(2), IAP(2), DHCY24 and ATPIA1). CONCLUSIONS: Remarkable apoptosis was found in placental trophoblast cells of PIH patients. Significant decrease in genes with anti-apoptosis functions can result in the apoptosis of placental trophoblast cells and thus contributes to the pathogenesis of PIH.