PM<sub>2.5</sub>激活HIF-1α-NF-κB/VEGF通路对肺损伤的影响

doi:10.3877/cma.j.issn.1674-6902.2022.03.006

目的

分析HIF-1α在PM_2.5暴露致肺损伤中的影响。

方法

构建经气管滴注PM_2.5悬液诱导大鼠肺损伤的动物模型，通过蛋白质免疫印迹、病理切片、ROS与TUNEL染色和ELISA等方法，分析不同PM_2.5暴露剂量下HIF-1α表达和肺损伤情况；构建抑制HIF-1α表达的动物模型，通过蛋白质免疫印迹、免疫荧光和ELISA等方法探讨HIF-1α在PM_2.5暴露致肺损伤中的作用机制。

结果

PM_2.5暴露造成肺组织病理性损伤，提高肺组织ROS水平、细胞凋亡率和湿干比，促进支气管肺泡灌洗液中各类炎症细胞计数和炎症因子IL-6、TNF-α水平增高，激活肺组织HIF-1α的蛋白表达，且以上效应与PM_2.5暴露浓度相关；抑制HIF-1α可降低NF-κB表达，降低支气管肺泡灌洗液中炎症因子水平，改善肺组织炎症；抑制HIF-1α可降低VEGF表达，降低支气管肺泡灌洗液中白蛋白水平和肺组织湿干比，改善肺组织水肿。

结论

PM_2.5通过激活"HIF-1α-NF-κB-炎症细胞(AMs、NEUs)/炎症因子(IL-6、TNF-α)-肺组织炎症反应"和"HIF-1α-VEGF-肺血管通透性-肺水肿"两条路径加重肺组织炎症反应、肺血管通透性和肺水肿，导致肺损伤。

关键词: 肺损伤, 细颗粒物, 缺氧诱导因子-1, 核因子-κB, 血管内皮生长因子

Objective

To explore the role of HIF-1α in PM_2.5-induced lung injury.

Methods

The rat model of lung injury induced by tracheal aerosol PM_2.5 suspension was established, HIF-1α expression and lung injury under different PM_2.5 exposure doses were investigated by western blot, pathological section, ROS staining, TUNEL staining and ELISA. An animal model was constructed to inhibit HIF-1α expression, and the mechanism of HIF-1α in lung injury induced by PM_2.5 exposure was investigated by western blot, immunofluorescence and ELISA.

Results

PM_2.5 exposure causes pathological damage, increases ROS level, apoptosis rate and wet-dry ratio of lung tissue, promotes the count of various inflammatory cells and the levels of inflammatory factors IL-6 and TNF-α in BALF, and activates the protein expression of HIF-1α in lung tissue, the above effects are related to PM_2.5 exposure concentration. Inhibition of HIF-1α can reduce lung inflammation by decreasing NF-κB expression in lung tissue and the level of inflammatory factors in BALF. Inhibition of HIF-1α can reduce lung tissue edema by decreasing VEGF expression in lung tissue, albumin level in BALF and lung tissue wet-dry ratio.

Conclusion

PM_2.5 aggravates lung tissue inflammation, pulmonary vascular permeability and pulmonary edema by activating two pathways: HIF-1α-NF-κB-inflammatory cells(AMs, NEUs)/inflammatory factors(IL-6, TNF-α)-lung tissue inflammation and HIF-1α-VEGF-pulmonary vascular permeability-pulmonary edema.

Key words: Lung injury, Particulate matter, Hypoxia-inducible factors-1α, Nuclear factor-κB, Vascular enclothelial growth factor

图1 不同处理组肺组织HIF-1α表达情况。注：**，P<0.01

图2 A：不同处理组肺组织病理切片，注：a：PBS组；b：PM_2.5(0.375 mg/kg)组；c：PM_2.5(1.5 mg/kg)组；d：PM_2.5(6 mg/kg)组；e：PM_2.5(24 mg/kg)组；蓝色箭头：水肿；绿色箭头：肺泡壁增厚；黄色箭头：炎症细胞。B、C：不同处理组肺组织ROS、TUNEL染色，注：a：PBS组；b：PM_2.5(1.5 mg/kg)组；c：PM_2.5(6 mg/kg)组；d：PM_2.5(24 mg/kg)组。注：*，P<0.05，**，P<0.01

图3 A：不同处理组BALF中IL-6、TNF-α水平。B：不同处理组BALF中白细胞分类计数。C：不同处理组肺组织湿干重比。注：*，P<0.05，**，P<0.01

图5 A：不同处理组HIF-1α和P-p65免疫荧光染色及相对荧光强度，注：蓝色：DAPI，红色：HIF-1α，粉色：P-p65。B：不同处理组BALF中IL-6、TNF-α水平。注：*，P<0.05，**，P<0.01，n.s.表示无统计学差异

图4 不同处理组肺组织HIF-1α表达情况。注：**，P<0.01

图6 A：不同处理组HIF-1α和VEGF免疫荧光染色及相对荧光强度，注：蓝色：DAPI，红色：HIF-1α，绿色：VEGF。B：不同处理组BALF中白蛋白水平。C：不同处理组肺组织湿干重比。注：*，P<0.05，**，P<0.01，n.s.表示无统计学差异

1	Balmes JR. Household air pollution from domestic combustion of solid fuels and health[J]. J Allergy Clin Immunol, 2019，143(6): 1979-1987.
2	韩璐瑶，吴克坚，高永恒，等. 不同大气污染物对呼吸系统疾病门诊量的影响[J/CD]. 中华肺部疾病杂志(电子版), 2020，13(2): 229-235.
3	韩璐瑶，金发光. 西安市大气污染物影响人群健康的现状[J/CD]. 中华肺部疾病杂志(电子版), 2019, 12(6): 783-785.
4	丁世彬，高丽云，李玉春，等. 慢性PM_2.5暴露对C57BL/6J小鼠肺组织炎症和NLRP3炎性小体活性的影响[J]. 中国实验动物学报，2019, 27(4): 444-449.
5	Zhang Y, Wang S, Zhu J, et al. Effect of atmospheric PM_2.5 on expression levels of NF-kappaB genes and inflammatory cytokines regulated by NF-kappaB in human macrophage[J]. Inflammation, 2018, 41(3): 784-794.
6	Yang B, Guo J, Xiao C. Effect of PM_2.5 environmental pollution on rat lung[J]. Environ Sci Pollut Res Int, 2018, 25(36): 36136-36146.
7	Semenza GL, Wang GL. A nuclear factor induced by hypoxia via de novo protein synthesis binds to the human erythropoietin gene enhancer at a site required for transcriptional activation[J]. Mol Cell Biol, 1992, 12(12): 5447-5454.
8	Liu Z, Zhang B, Wang XB, et al. Hypertonicity contributes to seawater aspiration-induced lung injury: Role of hypoxia-inducible factor 1alpha[J]. Exp Lung Res, 2015, 41(6): 301-315.
9	Suresh MV, Ramakrishnan SK, Thomas B, et al. Activation of hypoxia-inducible factor-1alpha in type 2 alveolar epithelial cell is a major driver of acute inflammation following lung contusion[J]. Crit Care Med, 2014, 42(10): e642-e653.
10	Sun HD, Liu YJ, Chen J, et al. The pivotal role of HIF-1alpha in lung inflammatory injury induced by septic mesenteric lymph[J]. Biomed Pharmacother, 2017, 91: 476-484.
11	Zhao X, Jin Y, Li H, et al. Hypoxia-inducible factor 1 alpha contributes to pulmonary vascular dysfunction in lung ischemia-reperfusion injury[J]. Int J Clin Exp Pathol, 2014，7(6): 3081-3088.
12	Wu G, Xu G, Chen DW, et al. Hypoxia exacerbates inflammatory acute lung injury via the toll-like receptor 4 signaling pathway[J]. Front Immunol, 2018, 9: 1667.
13	Liang S, Ning R, Zhang J, et al. MiR-939-5p suppresses PM_2.5-induced endothelial injury via targeting HIF-1alpha in HAECs[J]. Nanotoxicology, 2021, 15(5): 706-720.
14	Dai J, Sun C, Yao Z, et al. Exposure to concentrated ambient fine particulate matter disrupts vascular endothelial cell barrier function via the IL-6/HIF-1alpha signaling pathway[J]. FEBS Open Bio, 2016, 6(7): 720-728.
15	Gao Y, Sun J, Dong C, et al. Extracellular vesicles derived from adipose mesenchymal stem cells alleviate PM_2.5-induced lung injury and pulmonary fibrosis[J]. Med Sci Monit, 2020, 26: e922782.
16	Li R, Kou X, Xie L, et al. Effects of ambient PM_2.5 on pathological injury, inflammation, oxidative stress, metabolic enzyme activity, and expression of c-fos and c-jun in lungs of rats[J]. Environ Sci Pollut Res Int, 2015, 22(24): 20167-20176.
17	Lee K, Zhang H, Qian D Z, et al. Acriflavine inhibits HIF-1 dimerization，tumor growth, and vascularization[J]. Proc Natl Acad Sci USA, 2009, 106(42): 17910-17915.
18	Wang GL, Jiang BH, Rue EA, et al. Hypoxia-inducible factor 1 is a basic-helix-loop-helix-PAS heterodimer regulated by cellular O₂ tension[J]. Proc Natl Acad Sci U S A, 1995, 92(12): 5510-5514.
19	Wang GL, Semenza GL. Characterization of hypoxia-inducible factor 1 and regulation of DNA binding activity by hypoxia[J]. J Biol Chem, 1993, 268(29): 21513-21518.
20	Jiang H, Huang Y, Xu H, et al. Inhibition of hypoxia inducible factor-1alpha ameliorates lung injury induced by trauma and hemorrhagic shock in rats[J]. Acta Pharmacol Sin, 2012, 33(5): 635-643.
21	Rius J, Guma M, Schachtrup C, et al. NF-κB links innate immunity to the hypoxic response through transcriptional regulation of HIF-1α[J]. Nature, 2008, 453(7196): 807-811.
22	Jung Y, Isaacs JS, Lee S, et al. IL-1β mediated up-regulation of HIF-lα via an NFkB/COX-2 pathway identifies HIF-1 as a critical link between inflammation and oncogenesis[J]. FASEB J, 2003, 17(14): 1-22.
23	Diamant G, Dikstein R. Transcriptional control by NF-kappaB: elongation in focus[J]. Biochim Biophys Acta, 2013, 1829(9): 937-945.
24	Barboric M, Nissen RM, Kanazawa S, et al. NF-kappaB binds P-TEFb to stimulate transcriptional elongation by RNA polymerase Ⅱ[J]. Mol Cell, 2001, 8(2): 327-337.
25	Tan W, Jia W, Sun V, et al. Topical rapamycin suppresses the angiogenesis pathways induced by pulsed dye laser: molecular mechanisms of inhibition of regeneration and revascularization of photocoagulated cutaneous blood vessels[J]. Lasers Surg Med, 2012, 44(10): 796-804.
26	陈　觅，杨　扬，李欣宁，等. 血管内皮生长因子信号系统与急性肺损伤研究进展[J]. 国际麻醉学与复苏杂志，2021, 42(6): 668-672.
27	Powis G, Kirkpatrick L. Hypoxia inducible factor-1alpha as a cancer drug target[J]. Mol Cancer Ther, 2004, 3(5): 647-654.

[1]	李璐璐, 马利红, 金佳佳, 谷伟. 干扰素基因刺激因子通过肺巨噬细胞胞葬功能调控急性肺损伤小鼠修复的研究[J/OL]. 中华危重症医学杂志(电子版), 2024, 17(02): 97-103.
[2]	邵小丽, 林燕, 张玲玲, 韩亚琴. 超声引导下子宫肌瘤注射聚桂醇硬化术联合术后米非司酮治疗临床疗效分析[J/OL]. 中华妇幼临床医学杂志(电子版), 2024, 20(03): 353-360.
[3]	雷子威, 凌萍, 沈纵, 魏晨如, 朱邦晖, 伍国胜, 孙瑜. 类器官肺损伤疾病模型构建及应用的研究进展[J/OL]. 中华损伤与修复杂志(电子版), 2024, 19(06): 531-535.
[4]	黄福, 王黔, 金相任, 唐云川. VEGFR2、miR-27a-5p在胃癌组织中的表达与临床病理参数及预后的关系研究[J/OL]. 中华普外科手术学杂志(电子版), 2024, 18(05): 558-561.
[5]	朱佳琳, 方向, 贵诗雨, 黄丹, 周小雨, 郭文恺. 大鼠切口疝腹膜前间隙补片修补术后血清中VEGF 和Ang-1 的表达情况[J/OL]. 中华疝和腹壁外科杂志(电子版), 2024, 18(06): 703-707.
[6]	张敏龙, 杨翠平, 王博, 崔云杰, 金发光. MiR-200b-3p 通过抑制HIF-1α 表达减轻海水吸入诱导的肺水肿作用及机制[J/OL]. 中华肺部疾病杂志(电子版), 2024, 17(05): 696-700.
[7]	井发红, 李丽娜, 高婷, 高艳梅, 杨楠, 李卓, 慕玉东. 肺癌立体定向放疗血清SAP 和MMPs 表达及临床意义[J/OL]. 中华肺部疾病杂志(电子版), 2024, 17(05): 707-713.
[8]	张桂萍, 丘勇林, 湛绮婷, 孙乐栋. 晚期非小细胞肺癌血清Ape1/Ref-1对放射性肺损伤发生的预测意义[J/OL]. 中华肺部疾病杂志(电子版), 2024, 17(04): 519-523.
[9]	廖江荣, 吴秀琳, 陈光春, 郭亮, 吕慈, 蔡俊, 陈夕. 急性主动脉夹层并发急性肺损伤的研究新进展[J/OL]. 中华肺部疾病杂志(电子版), 2024, 17(03): 488-492.
[10]	顾晓凌, 吴冠楠, 宋勇. 核因子E2相关因子2(Nrf2)与铁死亡在脓毒症相关急性肺损伤中的研究进展[J/OL]. 中华肺部疾病杂志(电子版), 2024, 17(02): 324-328.
[11]	李玉娟, 艾芳, 熊欢庆, 陈键, 刘刚, 李志超, 金发光. "丹蛇"组方对小鼠急性肺损伤的治疗作用[J/OL]. 中华肺部疾病杂志(电子版), 2024, 17(02): 171-177.
[12]	林玲, 李京儒, 沈瑞华, 林惠, 乔晞. 基于生物信息学分析小鼠急性肾损伤和急性肺损伤的枢纽基因[J/OL]. 中华肾病研究电子杂志, 2024, 13(03): 134-144.
[13]	刘婷, 杨少康, 陈亿霏, 刘悦, 潘纯. 气道闭合的监测在机械通气中的研究进展[J/OL]. 中华重症医学电子杂志, 2024, 10(04): 394-398.
[14]	孙晓桐, 何怀武. 非对称性肺损伤的诊疗进展[J/OL]. 中华重症医学电子杂志, 2024, 10(03): 287-291.
[15]	刘建, 王文珠, 王倩. 老年髋部骨折术后肺损伤现状调查分析及影响因素研究[J/OL]. 中华诊断学电子杂志, 2024, 12(04): 260-264.

阅读次数

全文

摘要

选择文件类型/文献管理软件名称

选择包含的内容

PM_2.5 aggravates lung injury by activating HIF-1α-NF-κB/VEGF pathway

模态框（Modal）标题

选择文件类型/文献管理软件名称

选择包含的内容

PM2.5 aggravates lung injury by activating HIF-1α-NF-κB/VEGF pathway

PM_2.5 aggravates lung injury by activating HIF-1α-NF-κB/VEGF pathway