本文選題：胎糞吸入綜合征 + 法尼醇X受體��； 參考：《第三軍醫(yī)大學(xué)》2017年碩士論文

【摘要】：背景與目的胎糞吸入綜合征(Meconium aspiration syndrome,MAS)是指新生兒在子宮內(nèi)或產(chǎn)時(shí)吸入胎糞污染的羊水,導(dǎo)致氣道阻塞及全身炎癥反應(yīng),可累及全身各個(gè)臟器及系統(tǒng),癥狀因羊水污染程度及吸入的羊水量而表現(xiàn)不同。MAS發(fā)病機(jī)制復(fù)雜,尚未完全明確,重癥MAS并發(fā)癥多,治療困難,故MAS仍是產(chǎn)科醫(yī)師及新生兒科醫(yī)師重點(diǎn)關(guān)注疾病。細(xì)胞死亡是生命現(xiàn)象不可逆停止以及生命的結(jié)束,是細(xì)胞代謝的生物學(xué)過(guò)程,與細(xì)胞生長(zhǎng)、增殖、分化一樣都受到機(jī)體調(diào)控。前期實(shí)驗(yàn)證明:膽酸可誘導(dǎo)A549細(xì)胞死亡,低濃度膽酸誘導(dǎo)細(xì)胞凋亡,高濃度的膽酸誘導(dǎo)壞死性凋亡。但其具體的作用機(jī)制如何?我們將采用人肺癌上皮細(xì)胞株A549細(xì)胞作為工具細(xì)胞,A549細(xì)胞與肺泡上皮細(xì)胞功能相似,進(jìn)一步探討肺損傷因素如胎糞、膽酸等如何誘導(dǎo)細(xì)胞死亡。法尼醇X受體(Farnesoid X Receptor,FXR)是幾個(gè)代謝途徑的重要監(jiān)管者。FXR作為一種膽汁酸受體和膽汁酸生物合成的生物感受器,有助于影響膽汁酸在新陳代謝、炎癥及細(xì)胞周期控制中的作用。近年來(lái),多項(xiàng)臨床研究發(fā)現(xiàn)FXR在急性肺損傷(Acute Lung Injury,ALI)/急性呼吸窘迫綜合征(Acute Respiratory Distress Syndrome,ARDS)發(fā)生中發(fā)揮重要作用,但原因不清楚。前期有研究中發(fā)現(xiàn)FXR可能作為膽酸的核受體介導(dǎo)了膽酸誘導(dǎo)肺泡上皮細(xì)胞(Alveolar Epithelial Cells,AEC)死亡和抑制PS分泌的作用。具有調(diào)節(jié)炎癥及保護(hù)內(nèi)皮細(xì)胞功能的FXR為何在新生兒呼吸系統(tǒng)疾病中發(fā)揮損傷放大效應(yīng),其機(jī)制有待深入研究。方法第一部分:通過(guò)氣管插管,氣管內(nèi)注入胎糞制備新生大鼠胎糞吸入綜合征的動(dòng)物模型;第二部分:采用不同濃度胎糞混懸液刺激A549細(xì)胞,檢測(cè)細(xì)胞凋亡及壞死情況,并采用Wester blot免疫印跡法、SDS-PAGE凝膠電泳法測(cè)定經(jīng)胎糞刺激后的肺組織及細(xì)胞FXR及RIPK3的表達(dá)。探索FXR在新生大鼠胎糞吸入綜合征發(fā)病機(jī)制中的作用;第三部分:臨床選擇機(jī)械通氣新生兒,分為肺源性插管患者組、非肺源性插管患者的對(duì)照組,收集肺泡灌洗液細(xì)胞行流式細(xì)胞學(xué)檢查,分析肺泡細(xì)胞死亡比例及類型。結(jié)果(一)氣管導(dǎo)管內(nèi)注入胎糞混懸液2ml/kg的MAS組出現(xiàn)了相應(yīng)的肺損傷。(二)1.A549細(xì)胞經(jīng)過(guò)不同濃度的胎糞刺激后,細(xì)胞死亡比例明顯增加,呈濃度依賴性,胎糞混懸液濃度越高,A549細(xì)胞死亡比例越高。2.MAS與C組和NS組比較,MAS組動(dòng)物肺組織FXR及RIPK3蛋白表達(dá)水平有增加趨勢(shì)。3.不同濃度胎糞刺激A549細(xì)胞后,FXR、RIPK3的蛋白表達(dá)水平增加。(三)1.肺源性氣管插管組細(xì)胞死亡平均數(shù)為25%(凋亡21%)左右,非肺源性插管患者細(xì)胞死亡平均數(shù)為5%左右。2.肺源性氣管插管組血液中IL-6的平均值為533,明顯高于非肺源性插管患者。結(jié)論1.通過(guò)氣管導(dǎo)管內(nèi)注入胎糞混懸液2ml/kg可成功制作胎糞吸入綜合征模型,為研究胎糞吸入綜合征及其相關(guān)疾病提供更好的動(dòng)物模型。2.胎糞可誘導(dǎo)A549細(xì)胞死亡比例增加,呈濃度依賴性,20mg/ml組刺激24小時(shí)為最適濃度組,細(xì)胞死亡比例在39%左右。隨著胎糞混懸液濃度的增加,FXR、RIPK3的蛋白表達(dá)水平明顯增加。3.肺源性氣管插管組患者中,細(xì)胞死亡比例明顯大于非肺源性氣管插管患者,且死亡方式以凋亡為主。
[Abstract]:Background and objective Meconium aspiration syndrome (MAS) refers to the neonate inhalation of meconium contaminated amniotic fluid in the uterus or during birth, resulting in airway obstruction and systemic inflammatory reaction, which may involve the various organs and systems of the whole body. The symptoms are complex because of the degree of amniotic fluid pollution and the amount of amniotic fluid inhaled. It is not completely clear that the complications of severe MAS are many and the treatment is difficult, so MAS is still an obstetrician and newborn pediatrician. Cell death is an irreversible stop of life phenomenon and the end of life. It is the biological process of cell metabolism, which is controlled by the organism as well as cell growth, proliferation and differentiation. Can induce A549 cell death, low concentration of cholic acid to induce apoptosis and high concentration of cholic acid to induce necrotic apoptosis. But how is its specific mechanism? We will use human lung cancer epithelial cell line A549 cells as tool cells, A549 cells similar to alveolar epithelial cells, and further explore lung injury factors such as meconium, cholic acid and so on. What induces cell death. Farnesoid X Receptor (FXR) is an important regulator of several metabolic pathways,.FXR, as a biosynthesis of bile acid receptor and bile acid biosynthesis, which contributes to the effect of bile acids on metabolism, inflammation and cell cycle control. In recent years, a number of clinical studies have found FXR in a number of clinical studies. The cause of acute lung injury (Acute Lung Injury, ALI) / acute respiratory distress syndrome (Acute Respiratory Distress Syndrome, ARDS) plays an important role, but the reason is unclear. In previous studies, it was found that FXR may be used as a nuclear receptor for cholic acid to induce cholic acid induced alveolar epithelial cells (Alveolar Epithelial) death and inhibition. The role of secretory function. The mechanism of FXR, which regulates inflammation and protecting endothelial cell function, plays a damaging magnification effect in neonatal respiratory diseases. The first part: the first part: the animal model of neonatal meconium inhalation syndrome was prepared by intratratracheal intubation and intratracheal injection of meconium; the second part: different The concentration of meconium suspension was used to stimulate A549 cells to detect cell apoptosis and necrosis. The Wester blot immunoblotting method was used to determine the expression of FXR and RIPK3 in the lung tissues and cells stimulated by meconium by SDS-PAGE gel electrophoresis. The role of FXR in the pathogenesis of neonatal meconium inhalation syndrome was explored. The third part: clinical selection of machinery. The newborns were divided into pulmonary intubation group, non pulmonary intubation group and control group without pulmonary intubation. The cell line flow cytology of alveolar lavage fluid was collected to analyze the proportion and type of alveolar cell death. Results (1) MAS group of 2ml/kg in the endotracheal tube injected with meconium suspension showed corresponding lung injury. (two) 1.A549 cells passed different concentrations. After the meconium stimulation, the proportion of cell death increased significantly, the concentration depended, the higher the concentration of meconium suspension, the higher the proportion of A549 cell death, the higher the ratio of.2.MAS to C and NS, the expression level of FXR and RIPK3 protein in the lung tissue of MAS group was increased, and.3. at different concentration of meconium, the FXR, RIPK3 protein expression level was increased. (three) the average number of cell deaths in the 1. pulmonary endotracheal intubation group was 25% (apoptosis 21%), and the average number of cell deaths in non pulmonary intubation patients was 5%.2. pulmonary endotracheal intubation group, the mean value of IL-6 in the blood of pulmonary endotracheal intubation group was 533, obviously higher than that of non pulmonary intubation patients. Conclusion 1. through intratracheal catheter injection of meconium suspension 2ml/kg can be successfully made. The meconium aspiration syndrome model provides a better animal model for the study of meconium aspiration syndrome and related diseases..2. meconium can induce the increase of A549 cell death ratio, which is concentration dependent. The 20mg/ml group is stimulated for 24 hours as the optimum concentration group, the proportion of cell death is about 39%. With the increase of meconium suspension concentration, FXR, RIPK3 protein In the patients with.3. pulmonary endotracheal intubation, the proportion of cell death was significantly greater than that of non pulmonary endotracheal intubation patients, and the death mode was mainly apoptosis.
【學(xué)位授予單位】：第三軍醫(yī)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R722.1

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本文編號(hào)：2019013