The Mechanism of lncRNA NEAT1 in Alleviating Acute Respiratory Distress Syndrome Through miR-27b Regulated Nrf2 Pathway
试验速览
- 阶段
- 不适用
- 发起方
- 入组人数
- 425
- 试验地点
- 1
- 主要终点
- The expression of lncRNA NEAT1 in blood and BALF in all groups
研究概览
简要总结
The acute respiratory distress syndrome, formerly known as the acute lung injury (ARDS/ALI), is a critical illness with high mortality due to the lack of effective treatment. The pathogenesis of ARDS/ALI has not been fully elucidated. Nuclear factor E2-related factor 2 (Nrf2) plays a key role in regulating lung inflammation and oxidative stress which are closely related to lung injury in ARDS/ALI, but its regulatory mechanism remains unclear. The investigator's provious study shown that microRNA-27b (miR-27b) downregulated Nrf2 to aggravate lung inflammation and histological injury. Furthermore, in lipopolysaccharide (LPS)-induced cell (J774A.1) inflammation model, miR-27b was upregulated while the long non-coding RNA (lncRNA) NEAT1 was downregulated, the putative binding sites of lncRNA NEAT1 and miR-27b were successfully predicted by bioinformatics approach. Thus, the investigators propose that NEAT1 plays as a competing endogenous RNA (ceRNA) to adsorb miR-27b and liberate Nrf2, therefore, to attenuate lung inflammation and related lung injury in ARDS/ALI. This project aims to explore the role of the lncRNA NEAT1/ mir-27b /Nrf2 signal axis in the development and treatment of ARDS/ALI in patients, as well as in LPS-induced ALI animal and cell models by using bioinformatics, molecular biology, histomorphology and clinical phenotype approaches, and to clarify the new mechanism in ARDS/ALI development and to provide new therapeutic targets.
详细描述
Collect blood and BALF from 400 ARDS patients at different time (at check-in, 24, 48 and 72 h after check-in the hospital) and 25 gender and age matching healthy controls. Use RT-PCR to detect the expression of lncRNA NEAT1、miR-27b and Nrf2 in blood and BALF of ARDS patients and health controls. The expressions of inflammatory and oxidative stress associated factors (NLRP3、NF-κB-P65、 p-P65、IκB、p-IκB、HO-1、NQO1、caspase-1、IL-1β、IL-6、IL-18、TNF-α) will be detected by western blot、ELISA and RT-PCR. Moreover, flow cytometry will be adopted to measure the numbers and kinds of cells in BALF. Then, analyze the differences of the expressions of lncRNA NEAT1、miR-27b and Nrf2 in the groups. To explore the correlation of expressions of lncRNA NEAT1、miR-27b and Nrf2 with inflammation and oxidative stress in the groups. Finally, to declare the relative of lncRNA NEAT1、miR-27b and Nrf2 with the time of mechanical ventilation, severity and mortality in 28 days of ARDS patients.
研究设计
- 研究类型
- Observational
- 观察模型
- Other
- 时间视角
- Prospective
入排标准
- 年龄范围
- 18 Years 至 70 Years（Adult, Older Adult）
- 性别
- All
- 接受健康志愿者
- 是
入选标准
- •We included patients with acute respiratory distress according to 2012 ARDS Berlin new definition (Acute Respiratory Distress Syndrome: The Berlin Definition. JAMA, 2012, 307(23):2526).
- •Acute or progressive dyspnea within 1 week with identify cause;
- •Chest radiograph/chest CT showed double lung infiltration, which could not be fully explained by pleural effusion, atelectasis, or nodules;
- •Respiratory failure cannot be fully explained by heart failure and fluid overload;
- •Hypoxemia, partial pressure of oxygen in arterial blood (PaO2)/oxygen fraction in air (FIO2) <150 mm Hg under PEEP ≥5 cm H2O, (mild ARDS: 200mmHg<PaO2/FiO2≤300mmHg, moderate ARDS: 100mmHg<PaO2/FiO2≤200mmHg, severe ARDS: PaO2/FiO2≤100mmHg);
- •18~70 years old;
- •Agree to participate in the trial, and sign the informed consent.
排除标准
- •Age less than 18 years old;
- •Time of hospital stay <24 h;
- •Pregnancy;
- •Using V-V ECOM;
- •Cardiac index <1.5L·ml.min-1.m-2;
- •Pulmonary resection;
- •Pulmonary embolism ;
- •Refused to participate in the study.
结局指标
主要结局
The expression of lncRNA NEAT1 in blood and BALF in all groups
时间窗: up to 24 day
Use RT-PCR to measure the expression of lncRNA NEAT1 in blood and BALF in all groups
The expression of miR-27b in blood and BALF in all groups
时间窗: up to 3 day
Use RT-PCR to measure the expression of miR-27b in blood and BALF in all groups
The expression of Nrf2 in blood and BALF in all groups
时间窗: up to 3 day
Use RT-PCR and Wsetern blot to measure the expression of Nrf2 in blood and BALF in all groups
The expression of inflammatory factors(IL-1β、IL-6、IL-18、TNF-α) in blood and BALF in all groups
时间窗: up to 3 day
Use RT-PCR and ELISA to measure the expression of inflammatory factors(IL-1β、IL-6、IL-18、TNF-α) in blood and BALF in all groups
The expression of oxidative stress associated factors in blood and BALF in all groups
时间窗: up to 3 day
Use Western blot to measure the expression of oxidative stress associated factors(NLRP3、NF-κB-P65、 p-P65、IκB、p-IκB、HO-1、NQO1、caspase-1) in blood and BALF in all groups
The kinds of inflammatory cells in BALF and blood in all groups
时间窗: up to 3 day
Use flow cytometry to detect the kinds of inflammatory cells(neutrophile、macrophage、 lymphocyte) in BALF and blood in all groups
The time of mechanical ventilation of patients in ARDS groups
时间窗: up to28 day
Record the time of mechanical ventilation of patients in ARDS groups
The severity of ARDS patients in ARDS groups
时间窗: up to 28 day
Record the severity(PaO2/FiO2、OI、S/F、OSI) of ARDS patients in ARDS groups
The numbers and kinds of inflammatory cells in BALF and blood in all groups
时间窗: up to 3 day
Use flow cytometry to detect the number of inflammatory cells in BALF and blood in all groups
the mortality in 28 days of ARDS patients
时间窗: up to 28 day
Record the mortality in 28 days of ARDS patients
次要结局
- The differences and correlation of the expressions of lncRNA NEAT1、miR-27b and Nrf2 in the groups(up to 28 day)
- The correlation of expressions of lncRNA NEAT1、miR-27b and Nrf2 with inflammation and oxidative stress in the groups.(up to 28 day)
- The relative of lncRNA NEAT1、miR-27b and Nrf2 with the time of mechanical ventilation, severity and mortality in 28 days of ARDS patients(up to 28 day)
研究者
Guangfa Zhu
Director, Head of Respiratory and Critical Medicine Department
Beijing Anzhen Hospital