血浆microRNA与ARDS患者病情严重程度及预后相关性研究

发布时间：2018-05-10 11:52

本文选题：急性呼吸窘迫综合征(ARDS) + 肺内源性急性呼吸窘迫综合征(ARDSp)　；参考：《东南大学》2017年硕士论文

【摘要】：第一部分:血浆microRNA与ARDS患者病情严重程度及预后相关性的研究目的:本实验研究血浆中MSC-VEC-miRNA(与间充质干细胞及血管内皮细胞相关且既往在ARDS中有过研究的miRNA)水平与ARDS患者病情严重程度的相关性及其对患者预后的预测价值。方法:1、MSC-VEC-miRNA筛选:利用Pubmed数据库通过关键词及主题词检索相关文献,阅读文献筛选出MSC-VEC-miRNA。2、患者入组,标本采集:纳入2016年01月至2016年09月收住东南大学附属中大医院重症医学科诊断为ARDS且符合研究纳入和排除标准的患者,于患者诊断ARDS24小时内采外周血,离心后留取血浆保存。3、患者资料收集:基本资料(年龄、身高、体重等);一般情况(诊断、APACHE Ⅱ评分、SOFA评分、ARDS病因等);氧疗方式及参数(无创呼吸机、有创呼吸机及参数);预后指标(28天病死率等)。4、血浆miRNA检测:荧光定量PCR检测血浆中MSC-VEC-miRNA的循环阈值(CT值)。5、血浆内皮损伤标志蛋白及炎症因子检测:采用ELISA法检测患者血浆中VCAM-1、vWF、TNF-α、IL-10 浓度。结果:1、筛选 MSC-VEC-miRNA,符合条件的 MSC-VEC-miRNA 有14种:miR-15a,miR-16,miR-21,miR-24,miR-26a,miR-27a,miR-27b,miR-126,miR-146a,miR-150,miR-155,miR-221,miR-223,miR-320。2、存活组与死亡组患者一般情况比较:57例患者纳入研究分析,存活组39例,死亡组18例,28天病死率为31.6%。存活组与死亡组间比较:年龄、BMI、基础疾病、ARDS病因、呼气末正压(PEEP)、P02/Fi02无统计学差异(P0.05)。死亡组患者APACHE Ⅱ评分(26.9±8.5)、SOFA评分(13.7±3.9)和血乳酸(3.1[1.2-5.2])显著高于存活组(18.7±7.1,P0.001)、(8.9±4.5,P0.001)、(1.7[0.9-2.4],P=0.01)。而死亡组患者Murray评分显著高于存活组(2.8[2.3-3.7]vs.2.3[1.7-2.7],P=0.01)。3、存活组与死亡组患者血浆MSC-VEC-miRNA、血管内皮损伤蛋白及炎症因子比较:ARDS死亡组患者血浆miR-26a水平明显低于存活组(0.33[0.09-1.17]vs.0.97[0.17-3.49],P=0.046),而 ARDS 死亡组患者血浆 miR-320水平明显高于存活组(0.37[0.16-1.66]vs.0.18[0.07-0.39],P=0.041),其他 12 种miRNA在存活组与死亡组血浆中水平无统计学差异(P0.05)。存活组与死亡组血浆 vWF、VCAM-1、TNF-α、IL-10 浓度无统计学差异(P0.05)。4、不同ARDS严重程度患者血浆MSC-VEC-miRNA、血管内皮损伤蛋白及炎症因子比较:纳入患者中,ARDS轻度13例,中度25例,重度19例,轻、中、重度ARDS三组患者血浆MSC-VEC-miRNA水平无统计学差异(P0.05);三组患者血浆vWF、VCAM-1、TNF-α、IL-10水平无统计学差异(P0.05)。5、血浆MSC-miRNA对ARDS28天预后的预测价值:(1)miR-26a:AUC=0.67,p0.05;(2)miR-320:AUC=0.67,p0.05;(3)联合 miR-26a、miR-320:AUC=0.75,p0.001。结论:血浆MSC-VEC-miRNA水平与ARDS严重程度无相关性,血浆miR-26a、miR-320对ARDS患者28天预后有一定的预测价值。第二部分:肺内源性ARDS和肺外源性ARDS患者血浆microRNA表达水平差异的研究目的:本实验探究MSC-VEC-miRNA(与间充质干细胞及血管内皮细胞相关且既往在ARDS中有过研究的miRNA)在肺内源性ARDS和肺外源性ARDS患者血浆中水平差异。方法:1、MSC-VEC-miRNA筛选:利用Pubmed数据库通过关键词及主题词检索相关文献,通过阅读文献筛选出MSC-VEC-miRNA。2、患者入组,标本采集:纳入2016年01月至2016年09月收住东南大学附属中大医院重症医学科诊断为ARDS患者并符合本研究纳入和排除标准的患者57例,于患者诊断ARDS 24小时内采外周血,离心后留取血浆并保存。3、患者资料收集:基本资料(年龄、身高、体重等);一般情况(诊断、APACHE Ⅱ评分、SOFA评分、ARDS病因等);氧疗方式及参数(无创呼吸机、有创呼吸机及参数);预后指标(28天病死率等)。4、血浆miRNA检测:荧光定量PCR检测出血浆中MSC-VEC-miRNA的循环阈值(CT值)。5、血浆内皮损伤标志蛋白及炎症因子检测:采用ELISA法检测患者血浆中VCAM-1、vWF、TNF-α、IL-10浓度。结果:1、筛选 MSC-VEC-miRNA,符合条件 MSC-VEC-miRNA有14种:miR-15a,miR-16,miR-21,miR-24,miR-26a,miR-27a,miR-27b,miR-126,miR-146a,miR-150,miR-155,miR-221,miR-223,miR-320。2、肺内与肺外源性ARDS患者临床病情指标比较:符合标准的57例ARDS患者纳入研究分析,其中肺内源性ARDS 43例,肺外源性ARDS 14例。肺内源性ARDS患者与肺外源性ARDS患者一般情况比较:两组间年龄、性别、BMI、基础疾病、28天病死率、APACHE Ⅱ评分、SOFA评分、乳酸无统计学差异(P0.05)。肺内源性ARDS患者与肺外源性ARDS患者ARDS严重程度比较:PO2/FiO2在肺内源性ARDS组明显低于肺外源性ARDS组(145[119-203]vs.206[184-253],P=0.012);Murray肺损伤评分在肺内源性ARDS组显著高于肺外源性 ARDS 组(2.7[2-3.3]vs.1.8[1.3-2.4],P=0.008),两组间 FiO2、PEEP无统计学差异(P0.05)。3、肺内与肺外源性ARDS患者血浆MSC-VEC-miRNA比较:肺外源性ARDS 患者血浆肺外源性 ARDS 患者血浆 miR-221(0.22[0.12-0.49])、miR-27b(0.34[0.10-0.46])水平显著低于肺内源性 ARDS 患者(0.57[0.22-1.57],P=0.008)、(0.60[0.20-1.46],P=0.025),其他miRNA在两组患者血浆中表达水平无明显差异(P0.05)。4、肺内与肺外源性ARDS患者血管内皮损伤蛋白及炎症因子比较:肺外源性ARDS组患者血浆vWF显著低于肺内源性ARDS患者(0.77[0.29-1.54]vs.1.80[0.95-3.51],P=0.048);VCAM-1、IL-10、TNF-α 在两组患者血浆中的浓度无统计学差异(P0.05)。结论:miR-221、miR-27b、vWF在肺外源性ARDS患者血浆水平显著低于肺内源性ARDS患者。
[Abstract]:Part one: Study of the correlation between the severity and prognosis of patients with plasma microRNA and ARDS. Objective: To study the correlation between the level of plasma MSC-VEC-miRNA (related to mesenchymal stem cells and vascular endothelial cells and the previously studied miRNA in ARDS) and the severity of the severity of the patients with ARDS and the prognosis of the patients Methods: 1, MSC-VEC-miRNA screening: using the Pubmed database to retrieve the relevant literature through the key words and subject words, and read the literature to screen out MSC-VEC-miRNA.2, the patients into the group, the specimen collection: from 01 months to 09 months of 2016, the medical department of Zhongda Hospital Affiliated to Southeast University in the severe medical department was diagnosed as ARDS and accords with the inclusion and exclusion criteria. Patients received peripheral blood in ARDS24 hours and retained plasma after centrifugation and retained.3 after centrifugation. The data were collected: basic data (age, height, weight, etc.); general conditions (diagnosis, APACHE II score, SOFA score, ARDS etiology, etc.); oxygen therapy and parameters (non-invasive ventilator, invasive ventilator and parameters); prognosis index (28 days fatality rate, etc.). 4, plasma miRNA detection: fluorescence quantitative PCR detection of plasma MSC-VEC-miRNA cycle threshold (CT value).5, plasma endothelial damage marker protein and inflammatory factors detection: ELISA method for detecting VCAM-1, vWF, TNF- alpha, IL-10 concentration in patients plasma. Results: 1, screening MSC-VEC-miRNA, 14 types of MSC-VEC-miRNA. R-24, miR-26a, miR-27a, miR-27b, miR-126, miR-146a, miR-150, miR-155, miR-221, miR-223, miR-320.2, the general comparison between the survival group and the death group: 57 cases were included in the study analysis, the survival group 39 cases, the death group 18 cases, the 28 day fatality rate were compared between the 31.6%. survival group and the death group: age, basic disease, etiology, end expiratory pressure ( PEEP), there was no statistical difference in P02/Fi02 (P0.05). The score of APACHE II in the death group was (26.9 + 8.5), the SOFA score (13.7 + 3.9) and blood lactic acid (3.1[1.2-5.2]) were significantly higher than that in the survival group (18.7 + 7.1, P0.001), (8.9 + 4.5, P0.001), and (1.7[0.9-2.4], P=0.01). Compared with the plasma MSC-VEC-miRNA, vascular endothelial damage protein and inflammatory factors in the survival group and the death group, the plasma miR-26a level of the patients in the ARDS death group was significantly lower than that in the survival group (0.33[0.09-1.17]vs.0.97[0.17-3.49], P=0.046), while the plasma miR-320 level in the ARDS death group was significantly higher than that in the survival group (0.37[0.16-1.66]vs.0.18[0.07-0.39], P=). 0.041) there was no significant difference in the plasma levels of the other 12 kinds of miRNA in the survival group and the death group (P0.05). The plasma vWF, VCAM-1, TNF-, and IL-10 concentrations in the survival group and the death group were not statistically different (P0.05).4, and the plasma MSC-VEC-miRNA, vascular endothelial damage protein and inflammatory factors in patients with different ARDS severity were compared: ARDS mild 13 was included in the patients. There were 25 cases of moderate, 19 severe, moderate, moderate, and severe ARDS three groups with no statistically significant difference in plasma MSC-VEC-miRNA (P0.05); the plasma vWF, VCAM-1, TNF- a, IL-10 levels were not statistically different (P0.05) in the three groups, and the predictive value of plasma MSC-miRNA to ARDS28 day prognosis: (1) miR-26a:AUC=0.67, 2); (3) union 26a, miR-320:AUC=0.75, p0.001. conclusion: there is no correlation between plasma MSC-VEC-miRNA level and ARDS severity. Plasma miR-26a and miR-320 have a certain predictive value for the 28 day prognosis of ARDS patients. The second part: Study on the difference of plasma microRNA expression level in pulmonary endogenous ARDS and pulmonary ARDS patients: this experiment explored MSC-VEC-miRNA (and between) The levels of plasma levels of miRNA in the endogenous ARDS and pulmonary exogenous ARDS patients in ARDS were related to endothelium derived stem cells and vascular endothelial cells. Methods: 1, MSC-VEC-miRNA screening: using the Pubmed database to retrieve the relevant literature through the key words and subject words, and to select the MSC-VEC-miRNA.2 and the patients into the group through the reading literature. Sample collection: 57 patients, diagnosed as ARDS patients in the Department of severe medicine of Zhongda Hospital Affiliated to Southeast University from 01 to 2016 2016, were diagnosed as patients and were eligible for the inclusion and exclusion of this study. The peripheral blood was collected within 24 hours of the diagnosis of ARDS, the plasma was retained after centrifugation and.3 was preserved, and the data were collected: basic data (age, height, body). Heavy wait); general conditions (diagnosis, APACHE II score, SOFA score, ARDS etiology, etc.); oxygen therapy and parameters (non-invasive ventilator, invasive ventilator and parameters); prognostic index (28 days of mortality, etc.).4, plasma miRNA detection: fluorescence quantitative PCR detection of MSC-VEC-miRNA cycle threshold (CT).5, plasma endothelial damage marker protein and inflammation ELISA method was used to detect VCAM-1, vWF, TNF- alpha and IL-10 concentration in patients' plasma. Results: 1, screening MSC-VEC-miRNA, miR-15a, miR-16, miR-21, miR-24, miR-26a. Index comparison: 57 cases of ARDS patients who met the standard were included in the study, including 43 pulmonary endogenous ARDS cases and 14 pulmonary ARDS cases. Pulmonary endogenous ARDS patients and pulmonary exogenous ARDS patients were compared: the two groups were age, sex, BMI, basic disease, 28 day mortality, APACHE II score, SOFA score, and no statistical difference in lactic acid (P0.05). Lung The severity of ARDS in endogenous ARDS patients and pulmonary exogenous ARDS patients was compared: PO2/FiO2 in endogenous ARDS group was significantly lower than that of pulmonary ARDS group (145[119-203]vs.206[184-253], P=0.012), and Murray lung injury score in the pulmonary endogenous ARDS group was significantly higher than that in the pulmonary ARDS group (2.7[2-3.3]vs.1.8[1.3-2.4], P=0.012). No statistical difference (P0.05).3, plasma MSC-VEC-miRNA in patients with pulmonary and pulmonary exogenous ARDS: plasma miR-221 (0.22[0.12-0.49]) and miR-27b (0.34[0.10-0.46]) in pulmonary exogenous ARDS patients were significantly lower than that of lung endogenous ARDS (0.57 [0.22-1.57]). There was no significant difference in plasma expression level in the two groups (P0.05).4. The vascular endothelial damage protein and inflammatory factors in the pulmonary and exogenous ARDS patients were compared: the plasma vWF of the pulmonary exogenous ARDS patients was significantly lower than that of the pulmonary endogenous ARDS patients (0.77[0.29-1.54]vs.1.80[0.95-3.51], P= 0.048); VCAM-1, IL-10, TNF- alpha was in the plasma of two groups of patients. There was no significant difference in concentration (P0.05). Conclusion: miR-221, miR-27b and vWF in plasma of patients with extrinsic ARDS are significantly lower than those in patients with endogenous ARDS.

【学位授予单位】：东南大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R563.8

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