The elucidation of the mechanism of abortion/ premature birth by the key enzymes (PLA2 and COX) in the arachidonic acid cascade

花生四烯酸级联中关键酶（PLA2和COX）阐明流产/早产的机制

• 批准号: 13671712
• 负责人: KOYAMA Masayasu
• 金额: $ 2.3万
• 依托单位: Osaka University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13671712/
• 关键词: oxytocin receptor; preterm labor; methylated DNA; uterine contraction; chorioamnionitis; transcription; prostaglandin; HNE(4-hysroxy-2-nonenal)

I have focused attention to phospholipase A2 (PLA2) which has been thought to be the key enzyme associated uterine contractions and an inflammatory reaction. I developed high sensitive an enzyme activity measurement system with high-speed chromatography and measured the activity of PLA2 in serum, amniotic fluid and amniotic membrane of a clinical premature labor patient. In addition, I measured the concentration of immunoreactive PLA2 with RIA method by a specific monoclonal antibody of this enzyme (gift from Shionogi Pharmaceutical). As a result, a large quantity of PLA2 was secreted in serum and amniotic fluid of a premature labor patient and group II PLA2 principally was proved to be secreted from amniotic cells of a premature labor patient in large quantities. The concentration of the immunoreactive group II PLA2 in serum and amnion related to the enzymatic activity and the system of measurement was able to be established. Since this measurement system proved very high **ensitivity … More with HPLC, this enzyme activity measurement corollary was more superior than other inflammatory protein as a biosensor of premature labor. Furthermore, I identified manifestation of PLA2 to a premature labor patient without clinical inflammation. This result suggests that inflammation resemblance or abnormality of enzyme groups of the arachidonic acid cascade participates in onset mechanism of premature labor.It is necessary to consider influence of the oxidation stress which guides arachidonic acid metabolism (phospholipase and cyclooxygenase) when thinking about placental infection and mechanism of premature labor. HNE (4-hydroxy-2-nonenal) is a representative lipid peroxide degradation product. Alkyl modified protein with HNE is easy to be accumulated in the tissue because of its difficulty of digestion and it might be a good marker of oxidative stress. I demonstrated that HNE is strongly expressed in the placenta with clinical inflammatory diseases and manifestation of Cox-2 mRNA in the chorionic villi was observed in addition with HNE by RY-PCR. I also demonstrated that prostaglandine E2 was clearly elevated in the culture media of chorionic tissue by the addition of HNE. Less

我把注意力集中在磷脂酶A2（PLA 2），它被认为是与子宫收缩和炎症反应相关的关键酶。本研究建立了一种高灵敏度的酶活性检测系统，并应用该系统对一名早产患者的血清、羊水和羊膜中的PLA 2活性进行了检测。此外，我用该酶的特异性单克隆抗体（由Shionogi Pharmaceutical赠送）用RIA法测定了免疫反应性PLA 2的浓度。结果，在早产患者的血清和羊水中分泌大量的PLA 2，并且证明II组PLA 2主要从早产患者的羊水细胞大量分泌。建立了与酶活性相关的血清和羊膜中Ⅱ型PLA_2的浓度及测定系统。由于该测量系统证明了非常高的 ** 灵敏度 ...更多信息 用高效液相色谱法测定酶活性的推论，作为早产的生物传感器，比其它炎性蛋白更具有上级优势。此外，我确定了PLA 2的表现，早产患者没有临床炎症。这一结果表明，花生四烯酸级联反应的酶群的炎症相似性或异常参与了早产的发生机制，在考虑胎盘感染和早产机制时，有必要考虑引导花生四烯酸代谢的氧化应激（磷脂酶和环氧合酶）的影响。HNE（4-羟基-2-壬烯醛）是代表性脂质过氧化物降解产物。HNE烷基化修饰的蛋白质不易被消化，易在组织中积累，可作为氧化应激的良好标志物。我证明了HNE在临床炎症性疾病的胎盘中强烈表达，并且通过RY-PCR观察到除了HNE之外，绒毛中考克斯-2 mRNA的表现。我还证明了在绒毛膜组织的培养基中加入HNE明显升高了泽兰定E2。少

项目成果

Shimoya, K., Hashimoto, K., Shimizu, T.et al.: "Cervical fluid oncofetal firbronectin as a predictor of early ectopic pregnancy Is it affected by blood contamination?"J Reprod Med. 47(8). 640-644 (2002)

Shimoya, K.、Hashimoto, K.、Shimizu, T.等人：“宫颈液癌胎儿纤连蛋白作为早期异位妊娠的预测因子，它是否受到血液污染的影响？”J Reprod Med。

Nakamura, H., Kimura, T., Nakajima, A., et al.: "Detection of oxidative stress in seminal plasma and fractionated sperm from subfertile male patients"Eur J Obstet Gyn R B. 105. 155-160 (2002)

Nakamura, H.、Kimura, T.、Nakajima, A.等人：“精浆和来自低生育力男性患者的分级精子中氧化应激的检测”Eur J Obstet Gyn R B. 105. 155-160 (2002)

Q.Zhang, K.Shimoya, et al.: "Detection of fractalkine in huanm seminal plasma and its role in infertile patients"Hum Reprod. 17(16). 1560-1564 (2002)

Q.Zhang、K.Shimoya 等人：“Huam 精浆中 fractalkine 的检测及其在不孕患者中的作用”Hum Reprod。

Zhang, Q., Shimoya, K., et al.: "production of secretory leukocyte protease inhibitor by human amniotic membrances and regulation of its concentration in amniotic fluid"Mol Hum reprod. 7(6). 573-579 (2001)

张，Q.，Shimoya，K.，等人：“人羊膜分泌性白细胞蛋白酶抑制剂的产生及其在羊水中浓度的调节”Mol Hum reprod。

Ota, Y., Shimoya, K., Zhang, Q.et al.: "The expression of secretory leukocyte protease inhibitor(SLPI) in the fallopian tube : SLPI protects the acrosome reaction of sperm from inhibitory effects of elastase"Hum Reprod. 17(10). 2517-2522 (2002)

Ota, Y.、Shimoya, K.、Zhang, Q.等：“分泌性白细胞蛋白酶抑制剂 (SLPI) 在输卵管中的表达：SLPI 保护精子的顶体反应免受弹性蛋白酶的抑制作用”Hum Reprod。