Objectives: To clone genes specifically expressed in the placenta of patients with preeclampsia. and to explain the mechanism in the etiopathology of preeclampsia. Methods: The placentae of preeclamptic and normotensi...Objectives: To clone genes specifically expressed in the placenta of patients with preeclampsia. and to explain the mechanism in the etiopathology of preeclampsia. Methods: The placentae of preeclamptic and normotensive subjects with pregnancy were used as models, and the eDNA Library was constructed and 20 differentially expressed fragments were cloned after a new version of PCR-based subtractive hybridization. The false positive clones were identified by reverse dot blot analysis. With one of the obtained gene taken as the probe, the placentas of 10 normal pregnant women and 10 preeclamptic patients were studied by using dot hybridization methods. Results: Six false positive clones were identified by reverse dot blot, and the rest 14 clones were identified as preeclampsia-related genes. These clones were sequenced, and analyzed with BLAST analysis system. Eleven of 14 clones were genes already known, among which one belongs to necdin family; the rest 3 were identified as novel genes. These 3 genes were acknowledged by GenBank, with the accession numbers AF2322 16, AF2322 17, AF233648. The results of dot hybridization using necdin gene as probe were as follows: (1) There was this mRNA in the placental tissues of normal pregnancy as well as in that of preeclampsia. (2) The intensity of transcription of this mRNA in the placental tissues of preeclampsia increased significantly compared with that of the normal pregnancy (P＜0.05). Conclusions: This study for the first time reported this group of genes, especially necdin-expressing gene, which are related to the etiopathology of preeclampsia. In addition, the overtranscription of necdin gene has been found in preeclampsia. it is helpful in further studies of the etiology of preeclampsia.展开更多
文摘目的:探讨血管内皮生长因子在大鼠子痫前期模型中的作用。方法:选择清洁级W istar大鼠,随机分为两组,每组14只。对照组于妊娠第7天皮下注射生理盐水;亚硝基左旋精氨酸甲酯(L-NAM E)组于妊娠第7天皮下注射一氧化氮合成酶(NOS)抑制剂L-NAM E,每日125 m g/kg体重。两组均于妊娠第8天每天测量大鼠尾动脉血压,若血压增高合并尿蛋白大于或等于(+)时,大鼠子痫前期模型确立。同时,测定两组大鼠血小板数值和血清VEGF浓度后,立即处死大鼠测胎鼠及胎盘重量;采用免疫组化法检测胎盘VEGF表达量。结果:L-NAM E组孕鼠出现类似子痫前期特征,表现为血压明显升高至(145.3±4.6)mmHg,尿蛋白含量增加为(814.3±57.5)m g/L,血小板数显著减少至(467.1±76.3)×109/L及胎鼠胎盘重量明显减轻;对照组孕鼠血压(101.4±7.6)mmHg,尿蛋白(398.2±82.3)m g/L,血小板计数(837.20±71.4)×109/L。两组比较差异均有显著性(P均<0.05)。对照组血清VEGF浓度明显大于L-NAM E组[(13.62±3.33 vs 8.71±2.42)ng/L],胎盘中的VEGF表达量对照组也明显高于L-NAM E组[(0.89±0.19 vs 0.58±0.12)OD],差异均有显著性(P<0.01)。结论:大鼠血清VEGF水平降低和胎盘表达VEGF量下降参与子痫前期的发生。
文摘Objectives: To clone genes specifically expressed in the placenta of patients with preeclampsia. and to explain the mechanism in the etiopathology of preeclampsia. Methods: The placentae of preeclamptic and normotensive subjects with pregnancy were used as models, and the eDNA Library was constructed and 20 differentially expressed fragments were cloned after a new version of PCR-based subtractive hybridization. The false positive clones were identified by reverse dot blot analysis. With one of the obtained gene taken as the probe, the placentas of 10 normal pregnant women and 10 preeclamptic patients were studied by using dot hybridization methods. Results: Six false positive clones were identified by reverse dot blot, and the rest 14 clones were identified as preeclampsia-related genes. These clones were sequenced, and analyzed with BLAST analysis system. Eleven of 14 clones were genes already known, among which one belongs to necdin family; the rest 3 were identified as novel genes. These 3 genes were acknowledged by GenBank, with the accession numbers AF2322 16, AF2322 17, AF233648. The results of dot hybridization using necdin gene as probe were as follows: (1) There was this mRNA in the placental tissues of normal pregnancy as well as in that of preeclampsia. (2) The intensity of transcription of this mRNA in the placental tissues of preeclampsia increased significantly compared with that of the normal pregnancy (P＜0.05). Conclusions: This study for the first time reported this group of genes, especially necdin-expressing gene, which are related to the etiopathology of preeclampsia. In addition, the overtranscription of necdin gene has been found in preeclampsia. it is helpful in further studies of the etiology of preeclampsia.
