七氟醚对离体重度子痫前期胎盘蛋白表达的影响

发布时间：2018-05-19 12:51

  本文选题：重度子痫前期 + sEng蛋白　； 参考：《浙江大学》2014年硕士论文

【摘要】：子痫前期(pre-eclampsia PE)多发生于妊娠20周后,以高血压和蛋白尿为主要临床表现,伴全身多器官功能损害或功能衰竭,严重者可出现抽搐、昏迷,甚至死亡。目前,诱发子痫前期的病因及病理机制仍是围产医学中研究的一个重要课题。因此,子痫前期中胎盘组织内蛋白表达的改变,是PE发生发展的关键性环节之一。 目前认为子痫前期的发展主要分为两个阶段。第一阶段是无临床症状期。其主要特点是孕早期胎盘发育异常导致胎盘缺血、缺氧并释放过量的胎盘物质进入母体血循环。此阶段并无明显临床表现。第二阶段是临床症状期,其主要表现为高血压、肾损害、蛋白尿甚至HELLP综合症(溶血、肝酶升高、血小板降低)、子痫和其他器官损害。 胎盘是孕期的特殊器官,对胎儿的生长和母体的孕期状况都有很大的影响。因此,孕期胎盘的病理状况与胎儿生长受限及子痫前期均有密切相关。通过大量研究已经认识到胎盘浅着床和促炎因子的产生在子痫前期发病中的起到关键作用。其发病机制在于胎盘缺氧,异常的抗血管生成因子如sFltl,sEng等从胎盘中释放,抑制血管内皮生长因子(VEGF)、胎盘生长因子(PIGF)的生理作用,从而导致子痫前期的发生。 此外,对那些严重的重度子痫前期病人,任何疼痛刺激或紧张情绪都可诱发重度子痫前期患者抽搐,危及母婴生命安全,因此必须终止妊娠来挽救患者的生命。而在终止妊娠中采用吸入全身麻醉是理想的选择,在众多吸入麻醉剂中,七氟醚是一种应用于产科麻醉和无痛分娩的吸入麻醉药。血气分配系数小,具有快速诱导和苏醒的优点,能迅速从孕妇和新生儿体内清除,对新生儿Apgar评分和脐带血气无明显影响。因此在产科领域具有良好的应用前景。我们通过七氟醚干预重度子痫前期胎盘组织来观察其对胎盘组织的sEng蛋白表达有无改善作用。 本研究分成两个部分：第一部分：七氟醚对离体重度子痫前期胎盘以及外周血中sEng蛋白表达的影响重度子痫前期是子痫前期的一种,是导致孕妇、胎儿和新生儿死亡的主要原因。本研究通过对重度子痫前期患者胎盘组织的分离培养并通过七氟醚干预干预后sEng表达变化的研究,探讨七氟醚对重度子痫前期患者胎盘sEng蛋白表达的影响。 研究目的： (1)通过酶联免疫法(ELISA)检测重症子痫前期孕妇外周血浆中sEng水平和脐血sEng水平有无差异；(2)运用3%七氟醚干预重症子痫前期产妇的胎盘组织,用酶联免疫法(ELISA)检测检测胎盘组织干预在干预前后的分泌的sEng蛋白的变化,探讨吸入七氟醚对重度子痫前期患者胎盘分泌sEng蛋白的影响,为子痫前期的治疗及麻醉方案提供一个新的方法和思路。 方法： 1.研究对象为2012年6月至2013年1月间在本院住院行择期剖宫产术的重度子痫前期患者48例,重度子痫前期的诊断标准参照《威姆斯产科学》,纳入标准为既往无高血压、心脏病、肾脏病、糖尿病及甲状腺功能亢进症等病史的病例。 2.胎盘组织培养24h后干预于1MAC七氟醚1h,检测干预前后胎盘组织分泌的sEng的浓度。sEng的检测用酶联免疫吸附分析法(Elisa) 结果： 1.脐血的sEng浓度高于母血血清sEng浓度(P0.05) 2.七氟醚干预前后的sEng表达有差异,与干预前相比,干预后的sEng表达下降(P0.05)。 结论： 1.sEng是引起子痫前期的主要因素,由于sEng主要来源于胎盘。而重度子痫前期胎盘组织经七氟醚干预后分泌的sEng减少,由此可以证明七氟醚对重度子痫前期症状的改善可能有一定的作用。 第二部分：免疫印迹法分析七氟醚对细胞骨架蛋白在离体重症子痫前期患者胎盘中滋养层细胞中表达的影响 研究目的：已有研究表明,keratin, type I cytoskeletal-9(细胞骨架蛋白-9)在子痫前期患者胎盘中的表达远高于正常胎盘组织,而相关机制的研究仍鲜有报道,故此实验旨在研究七氟醚对子痫前期患者胎盘中该蛋白表达的影响。 方法： 1、研究对象为2012年8月—2013年5月在浙江大学附属妇产科医院产科住院分娩的孕妇60例,分为重度子痫患者组40例、正常妊娠孕妇(对照)组20例,收集其胎盘组织。 2、将正常胎盘组织设为A组,重症子痫前期患者胎盘组织分为B、C两组。分别用37℃5%CO2-95%O2.37℃5%CO2-95%O2干预混合七氟醚气体分别干预处理。 3、应用Western blot检测A,B,C3组的胎盘组织中cytoskeletal-9表达的差异；从而观察七氟醚处理后的重症子痫前期产妇的胎盘组织内cytoskeletal-9表达的变化。 结果： 1、在重度子痫前期患者胎盘中,cytoskeletal-9表达与正常妊娠的胎盘相比显著升高(P0.01)；用七氟醚处理后,重度子痫前期患者胎盘中cytoskeletal-9的表达水平较未处理的组明显降低(P0.01),但和正常妊娠的胎盘相比仍偏高(P0.01)。 结论： 1、重症子痫前期患者胎盘组织中keratin, type I cytoskeletal-9(细胞骨架蛋白-9)表达量增加。 2、七氟醚能降低重度子痫前期患者胎盘中cytoskeletal-9的表达水平。
[Abstract]:Preeclampsia (pre-eclampsia PE) occurs more frequently after 20 weeks of pregnancy, with hypertension and proteinuria as the main clinical manifestation, multiple organ dysfunction or functional failure in the whole body. Severe convulsions, coma and even death can occur in severe cases. At present, the etiology and pathological mechanism of preeclampsia in the preeclampsia are still an important subject in perinatal medicine. The change of placental protein expression in preeclampsia is one of the key links in the development of PE.
At present, the development of preeclampsia is mainly divided into two stages. The first stage is no clinical symptom phase. Its main feature is placental ischemia in the early pregnancy, hypoxia and release of excessive placental material into the maternal blood circulation. There is no obvious clinical manifestation in this stage. The second stage is the clinical symptom period, its main manifestation is Hypertension, kidney damage, proteinuria and even HELLP syndrome (hemolysis, elevated liver enzymes, thrombocytopenia), eclampsia and other organ damage.
Placenta is a special organ during pregnancy, which has a great influence on the growth of the fetus and the status of maternal pregnancy. Therefore, the pathological status of placenta during pregnancy is closely related to fetal growth restriction and preeclampsia. Its pathogenesis lies in placenta anoxia. Abnormal antiangiogenic factors such as sFltl, sEng, etc. are released from the placenta and inhibit the physiological role of vascular endothelial growth factor (VEGF) and placental growth factor (PIGF), leading to the occurrence of preeclampsia.
In addition, for severe preeclampsia, any pain irritation or tension can induce convulsions in severe preeclampsia and endanger the safety of mother and infant life. Therefore, it is necessary to terminate pregnancy to save the life of the patient. In the termination of pregnancy, inhalation general anesthesia is an ideal choice, seven fluorine among a large number of inhaled anesthetics. Ether is a inhaled anesthetic used in obstetric and painless delivery. The coefficient of blood gas distribution is small and has the advantages of rapid induction and revival. It can be quickly removed from pregnant women and neonates, and has no obvious influence on the Apgar score of the newborn and the umbilical cord blood gas. Therefore, we have a good application prospect in the field of obstetrics. We use sevoflurane to intervene in the field of obstetrics. Severe preeclampsia placenta tissues were used to observe the effect of placental tissue on the expression of sEng protein.
This study is divided into two parts: the first part: the effect of sevoflurane on the expression of sEng protein in the placenta and peripheral blood of severe preeclampsia in vitro, severe preeclampsia is a preeclampsia, which is the main cause of death in pregnant women, fetus and newborn. This study was conducted by isolation and culture of placental tissue in preeclampsia patients. The effect of sevoflurane on the expression of sEng protein in placenta of severe preeclampsia was studied through intervention of sevoflurane intervention on the expression of sEng.
The purpose of the study is:
(1) detection of sEng level in peripheral plasma and sEng level of umbilical cord blood in pregnant women with severe preeclampsia by enzyme linked immunosorbent assay (ELISA); (2) use 3% sevoflurane to intervene in placental tissue of preeclampsia parturients and detect the changes of sEng protein secreted by placental tissue before and after intervention by enzyme linked immunosorbent assay (ELISA). The effect of sevoflurane on the secretion of sEng protein in placenta of severe preeclampsia patients provides a new method and train of thought for the treatment of preeclampsia and the anesthesia plan.
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