切换至 "中华医学电子期刊资源库"
高级检索
中华肺部疾病杂志(电子版)

中华肺部疾病杂志(电子版) ›› 2019, Vol. 12 ›› Issue (01) : 9 -14. doi: 10.3877/cma.j.issn.1674-6902.2019.01.002

所属专题： 文献；

论著

NLRP3炎症小体在海水吸入性急性肺损伤中的表达和作用
王虎1, 鲁曦1, 王博文2, 师少魁3, 柳松勃4, 金发光1,†()   
  1. 1. 710038　西安，空军军医大学（第四军医大学）唐都医院呼吸与危重症医学科
    2. 361001　厦门，厦门大学附属成功医院（解放军第174医院）
    3. 721004　宝鸡，中国人民解放军第三医院
    4. 710043　西安，西安市北方医院呼吸科
  • 收稿日期:2018-12-25 出版日期:2019-02-20
  • 通信作者: 金发光
  • 基金资助:
    军队保健项目(14BJZ55); 国家自然科学基金面上项目(81270124)

Expression and effect of NLRP3 inflammasome in acute lung injury induced by seawater

Hu Wang1, Xi Lu1, Bowen Wang2, Shaokui Shi3, Songbo Liu4, Faguang Jin1,†()   

  1. 1. Department of Respiratory diseases, Tangdu Hospital, the Air Force Military Medical University, Xi′an 710038, China
    2. Chenggong Hospital Affiliated to Xiamen University (No.174 Hospital of PLA )1, Xiamen 361001, China
    3. Department of Respiratory diseases, No.3 Hospital of PLA, Baoji 721004, China
    4. Department of Respiratory diseases, North Hospital, Xi′an 710043, China
  • Received:2018-12-25 Published:2019-02-20
  • Corresponding author: Faguang Jin
  • About author:
    Corresponding author: Jin Faguang, Email:
全文 ( ) 下载PDF ( ) 导出引用
引用本文：

王虎, 鲁曦, 王博文, 师少魁, 柳松勃, 金发光. NLRP3炎症小体在海水吸入性急性肺损伤中的表达和作用[J]. 中华肺部疾病杂志(电子版), 2019, 12(01): 9-14.

Hu Wang, Xi Lu, Bowen Wang, Shaokui Shi, Songbo Liu, Faguang Jin. Expression and effect of NLRP3 inflammasome in acute lung injury induced by seawater[J]. Chinese Journal of Lung Diseases(Electronic Edition), 2019, 12(01): 9-14.

目的

探讨海水吸入型急性肺损伤大鼠肺组织中NOD样受体热蛋白结构域相关蛋白3(NLRP3)炎症小体表达的变化及介导的炎性因子在急性肺损伤(ALI)发生发展中的作用。

方法

将50只健康雄性SD大鼠随机分为5组，对照组，海水吸入1 h组，海水吸入3 h组，海水吸入6 h组，海水吸入9 h组，每组10只。采用经气管缓慢滴注(3 ml/kg)海水的方法制作大鼠损伤模型。制作大鼠肺脏石蜡切片并HE染色观察病理形态学变化。检测大鼠肺组织湿干比。ELISA检测测定各组肺组织中IL-1β和IL-18水平，RT-PCR检测肺组织中IL-1β、IL-18和NLRP3mRNA的表达。Western-blot检测肺组织中NLRP3蛋白表达。

结果

气管滴注海水后成功复制海水吸入性急性肺损伤模型。肺组织湿干比较对照组显著升高。病理形态学观察可见肺组织大量炎细胞浸润、水肿、间质增厚。各组大鼠血清中IL-1β和IL-18的水平随着时间增加逐渐升高，且在3～6 h达到顶峰，随后炎症因子的表达逐渐降低。与空白对照组比较，差异均有统计学意义(均P<0.05)。各组大鼠肺组织中IL-1β和IL-18的mRNA的表达水平与大鼠肺组织中IL-1β和IL-18的表达基本一致。与空白对照组比较，差异均有统计学意义(均P<0.05)。肺组织匀浆中NLRP3转录和翻译结果显示海水吸入刺激后，肺组织中NLRP3的mRNA和NLRP3蛋白含量变化含量随着时间明显逐渐增加，差异均有统计学意义(均P<0.05)。

结论

海水刺激下，NLRP3炎症小体介导的炎症反应参与了急性肺损伤发病过程并加重了肺损伤的程度，可能是海水急性肺损伤的发病机制之一，但其作用有待进一步证实。

关键词: 急性肺损伤, 海水, NLRP3炎症小体
Objective

To investigate the expression and the role of NOD-like receptor thermal-protein domain related protein (NLRP3) in the lung tissues and its mediated inflammatory factors in the occurrence and development of acute lung injury (ALI).

Methods

50 healthy male SD rats were randomly divided into 5 groups: the control group, the seawater inhalation 1 h group, the seawater inhalation 3 h group, the seawater inhalation 6 h group and the seawater inhalation 9 h group, with 10 rats in each group. The rat injury model was established by the transtracheal seawater slow infusion (3 ml/kg). The rat lung paraffin sections were made and the pathomorphology was observed by HE staining. The wet/dry weight ratio of the lung tissue was detected in rats. The expressions of IL-1β and IL-18 in lung tissues were detected by ELISA. The expressions of IL-β, IL-18 and NLRP3 mRNA in lung tissue were detected by RT-PCR. The expression of NLRP3 protein in lung tissue was detected by Western-blot.

Results

The rat model of acute lung injury induced by seawater inhalation was successfully replicated. The wet/dry weight ratio of the lung tissue was significantly higher compared with the control group. There was a large number of inflammatory cell infiltration, edema and interstitial thickening in the lung tissue. The ELISA has revealed that the levels of IL-1β and IL-18 in the serum of rats gradually increase over time, reaching the peak in 3-6 hours, and then the expression of inflammatory factors gradually decreases. Compared with the results of the control group, the difference was statistically significant (P<0.05). The RT-PCR has found that the expression of mRNA of IL-1β and IL-18 in lung tissues of rats were similar to the expression of IL-1β and IL-18 in lung tissues of rats. Compared with results of the control group, the difference was statistically significant (P<0.05). Western Blot and RT-PCR have been used to detect the transcription and translation of NLRP3 in lung homogenate. The findings indicate that the mRNA and protein in NLRP3 in lung tissues increase gradually over time with the stimulation of seawater inhalation, and the difference was statistically significant (P<0.05).

Conclusion

NLRP3 inflammasome mediated inflammatory response may be involved in the pathogenesis of acute lung injury and aggravate the degree of lung injury, which may be one of the pathogenesis of acute lung injury induced by seawater. However, its role needs further confirmation.

Key words: Acute lung injury, Seawater, NLRP3 inflammasome
图1 大鼠肺组织湿干比检测结果；注：0H组：空白对照组；1H组：海水吸入1 h组；3H组：海水吸入3 h组；6H组：海水吸入6 h组；9H组：海水吸入9 h组；***P<0.001，**P<0.01，*P<0.05 vs. 0H组
图2 大鼠肺组织病理学检测检测结果；注：A：对照组；B：海水吸入1 h组；C：海水吸入3 h组；D：海水吸入6 h组(HE×40)
图3 大鼠组织中IL-1β和IL-18的表达水平；注：0H组：空白对照组；1H组：海水吸入1 h组；3H组：海水吸入3 h组；6H组：海水吸入6 h组；9H组：海水吸入9 h组；*** P<0.001，**P<0.01，*P<0.05 vs. 0H组
图4 大鼠组织中IL-1β和IL-18 mRNA的表达水平；注：0H组：空白对照组；1H组：海水吸入1 h组；3H组：海水吸入3 h组；6H组：海水吸入6 h组；9H组：海水吸入9 h组；**P<0.01，*P<0.05 vs. 0H组
图5 大鼠肺组织中NLRP3mRNA的表达水平；注：**P<0.01，*P<0.05 vs. 0 H组
图6 大鼠肺组织中NLRP3蛋白检测结果；注：**P<0.01，*P<0.05 vs. 0 H组
1
Salomez F,Vincent JL. Drowning: a review of epidemiology, pathophysiology，treatment and prevention[J]. Resuscitation, 2004, 63(3): 261-268.
2
Orlowski JP. Drowning, near-drowning, and ice-water drowning[J]. JAMA, 1988, 260(3): 390-391.
3
Orlowski JP. Drowning, near-drowning, and ice-water submersions[J]. Pediatr Clin North Am, 1987, 34(1): 75-92.
4
杨昱，王关嵩，邓朝霞，等. 海水淹溺型肺损伤与淡水淹溺型肺损伤特征的比较[J]. 解放军医学杂志，2009, 34(4): 393-396.
5
金发光. 急性肺损伤的诊治研究现状及进展[J/CD]. 中华肺部疾病杂志(电子版), 2013, 6(1): 1-3.
6
Bryan NB. Activation of inflammasomes requires intracellular redistribution of the apoptotic speck-like protein containing a caspase recruitment domain[J]. J Immunol, 2009, 182(5): 3173-3182.
7
Bauernfeind F,Ablasser A,Bartok E, et al. Inflammasomes: current understanding and open questions[J]. Cell Mol Life Sci, 2011, 68(5): 765-783.
8
Dolinay T,Kim Y,Howrylak J, et al. Inflammasome-regulated cytokines are critical mediators of acute lung injury[J]. Am J Respir Crit Care Med, 2012, 185(11): 1225-1234.
9
Domej W,Foldes-Papp Z,Flogel E, et al. Chronic obstructive pulmonary disease and oxidative stress[J]. Curr Pharm Biotechnol, 2006, 7(2): 117-123.
10
dos Santos CC. The role of the inflammasome in ventilator-induced lung injury[J]. Am J Respir Crit Care Med, 2012, 185(11): 1141-1144.
11
dos Santos G,Kutuzov MA,Ridge KM. The inflammasome in lung diseases[J]. Am J Physiol Lung Cell Mol Physiol, 2012, 303(8): L627-L633.
12
Xinmin D,Yunyou D,Chaosheng P, et al. Dexamethasone treatment attenuates early seawater instillation-induced acute lung injury in rabbits[J]. Pharmacol Res, 2006, 53(4): 372-379.
13
李佳欢，许敏，金发光. 海水淹溺性肺损伤中肺水肿发生机制的研究进展[J]. 国际呼吸杂志，2010, 30(14): 877-880.
14
李聪聪，薄丽艳，刘庆晴，等. 蛋白激酶SPAK在海水淹溺性肺损伤中的表达和作用[J/CD]. 中华肺部疾病杂志(电子版), 2014, 7(1): 19-22.
15
Folkesson HG,Kheradmand F,Matthay MA. The effect of salt water on alveolar epithelial barrier function[J]. Am J Respir Crit Care Med, 1994, 150(6 Pt 1): 1555-1563.
16
He DK,Shao YR,Zhang L, et al. Adenovirus-delivered angiopoietin-1 suppresses NF-kappaB and p38 MAPK and attenuates inflammatory responses in phosgene-induced acute lung injury[J]. Inhal Toxicol, 2014, 26(3): 185-192.
17
Martinon F,Burns K,Tschopp J. The inflammasome: a molecular platform triggering activation of inflammatory caspases and processing ofproIL-beta[J]. Mol Cell, 2002, 10(2): 417，426.
18
Hornung, V．，Bauernfeind, F．，Halle, A．，et al. (2008) Silica crystals and aluminum salts activate the NALP3 inflammasome through phagosomal destabilization[J]. Nat Immunol, 9：847-856.
19
Yazdi, A．，Guarda, G．，Riteau, N．，et al. (2010) Nanoparticles activate the NLR pyrin domain containing 3 (NLRP3)inflammasome and cause pulmonary inflammationthrough release of IL-1alpha and IL-1beta[J]. Proc Natl Acad Sci USA, 107: 19449-19454.
20
Latz E,Xiao TS,Stutz A. Activation and regulation of the inflammasomes[J]. Nat Rev Immunol, 2013, 13: 397-411.
21
De Nardo D,Latz E. NLRP3 inflammasomes link inflammation and metabolic disease[J]. Trends Immunol, 2011, 32: 373-379.
22
Davis BK,Wen H,Ting JP. The inflammasome NLRs in immunity, inflammation, and associated diseases[J]. Annu Rev Immunol, 2010, 29: 707-735.
23
Witzenrath, M．，Pache, F．，Lorenz, D．，et al. (2011) The NLRP3 inflammasome is differentially activated by pneumolysin variants and contributes to host defense in pneumococcal pneumonia[J]. J Immunol, 187: 434-440.
24
Martinon, F．，Petrilli, V．，Mayor, A．，et al. (2006) Gout-associated uric acid crystals activate the NALP3 inflammasome[J]. Nature, 440: 237-241.
25
Vande Walle L,Van Opdenbosch N,Jacques P, et al. Negative regulation of the NLRP3 inflammasome by A20 protects against arthritis[J]. Nature, 2014, 512: 69-73.
[1] 梁哲浩, 方明笋, 胡弘毅, 陶涛, 徐孝平, 孙华琴. 基于生物信息学分析筛选脓毒症诱导急性肺损伤的关键基因[J]. 中华危重症医学杂志(电子版), 2022, 15(05): 360-366.
[2] 赵希伟, 周佳伟, 刘凯, 侯林义, 张文凯. 连接蛋白43通过蛋白激酶A介导丝氨酸373调控脓毒症急性肺损伤肺泡Ⅱ型上皮细胞屏障功能的研究[J]. 中华危重症医学杂志(电子版), 2021, 14(05): 355-361.
[3] 沈纵, 魏晨如, 朱邦晖, 包郁露, 伍国胜, 孙瑜. 间充质干细胞治疗吸入性损伤的动物实验研究进展[J]. 中华损伤与修复杂志(电子版), 2023, 18(02): 180-183.
[4] 蔡维霞, 曹涛, 赵明, 肖丹, 贾艳慧, 王璟, 张月, 王克甲, 韩军涛, 胡大海. Notch信号通路对烧伤大鼠血清诱导的肺血管内皮细胞细胞间黏附分子-1的影响[J]. 中华损伤与修复杂志(电子版), 2022, 17(04): 292-299.
[5] 周强, 赵烨德, 王雨翔, 肖仕初. 烧伤合并烟雾吸入性肺损伤病理机制和治疗研究新进展[J]. 中华损伤与修复杂志(电子版), 2022, 17(02): 171-175.
[6] 熊欢庆, 李玉娟, 陈键, 刘刚, 李志超, 金发光. 丹参酮IIA及苦参碱组方对脂多糖致小鼠急性肺损伤的协同保护作用[J]. 中华肺部疾病杂志(电子版), 2023, 16(04): 455-459.
[7] 李埝, 赵建军, 张建勇, 赵睿桢. hAMSCs调控MAPK信号通路对急性肺损伤AQP1的影响[J]. 中华肺部疾病杂志(电子版), 2023, 16(02): 156-163.
[8] 刘欣, 李艳敏, 郑佳, 赵商岐, 唐晓慧, 赵静, 周文涛, 周晓涛. COPD患者NLRP3炎症小体及TNF-α、HMGB1的表达及相互关系[J]. 中华肺部疾病杂志(电子版), 2022, 15(04): 468-472.
[9] 朱冠能, 汪洋, 宋海苗, 汪骏东. 血清铁蛋白及C反应蛋白水平对胸部创伤后急性肺损伤的预测意义[J]. 中华肺部疾病杂志(电子版), 2022, 15(03): 379-381.
[10] 许发琼, 贺斌峰, 黄朝旺, 胡明冬. 非编码RNA调控巨噬细胞炎症反应在ALI/ARDS中的研究进展[J]. 中华肺部疾病杂志(电子版), 2021, 14(05): 677-680.
[11] 顾艳利, 宋勇, 张方. 姜黄素在肺部炎症性疾病中的免疫调节作用[J]. 中华肺部疾病杂志(电子版), 2021, 14(04): 539-542.
[12] 袁高洁, 李忠俊. 输血相关急性肺损伤的研究进展[J]. 中华肺部疾病杂志(电子版), 2021, 14(03): 397-399.
[13] 王守森, 徐永君, 张灏. 加强海战颅脑损伤合并海水相关性损伤的研究[J]. 中华神经创伤外科电子杂志, 2021, 07(05): 257-260.
[14] 陈梦婷, 孟潇潇, 王瑞兰. 急性肺损伤时肺部微环境介导的细胞代谢变化的研究进展[J]. 中华重症医学电子杂志, 2022, 08(01): 80-84.
[15] 胡俊晟, 黄荣, 黄毅, 曾光, 金永志, 李梦帆. 丹参多酚酸盐通过Nrf2/HO-1信号通路对脂多糖诱导的小鼠急性肺损伤的保护作用[J]. 中华临床医师杂志(电子版), 2021, 15(12): 1024-1030.
阅读次数
全文


摘要

版权所有 中华医学会 中华医学电子音像出版社有限责任公司  京ICP备14006079号-1  网络出版服务许可证：(署)网出证(京)字第075号