NO吸入对急性肺损伤大鼠肺水肿程度及TLR 4和TNF-α表达的影响
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摘要
目的观察吸入不同浓度一氧化氮(nitric oxide,NO)对大鼠肺水肿程度、气道Toll样受体4(toll-like receptor 4,TLR 4)、肿瘤坏死因子α(tumor necrosis factor alpha,TNF-α)表达的影响。方法健康雄性SD大鼠36只,随机分对照组、模型组、NO浓度10×10~(-6) mg/L组、NO浓度20×10~(-6) mg/L组、NO浓度40×10~(-6) mg/L组、NO浓度100×10~(-6) mg/L组,每组各6只大鼠。除对照组外其余5组均气管内滴注细菌脂多糖(lipopolysaccharide,LPS),建立大鼠急性肺损伤(acute lung injury,ALI)模型,观察大鼠的肺水肿程度,以及分别吸入10×10~(-6) mg/L,20×10~(-6) mg/L,40×10~(-6) mg/L,100×10~(-6) mg/L的NO对大鼠TLR 4及TNF-α表达的影响。结果结果显示TLR 4和TNF-α在对照组大鼠气道内有广泛分布和表达。模型组及各NO吸入组TLR 4及TNF-α表达明显升高,肺水肿程度明显高于对照组(P<0.05)。与模型组比较,NO 10×10~(-6) mg/L组、NO 20×10~(-6) mg/L组、NO 40×10~(-6) mg/L组的TLR 4及TNF-α表达量明显减少,肺水肿程度明显减轻(P均<0.05),但NO 100×10~(-6) mg/L组肺水肿程度明显加重(P<0.05), TLR 4及TNF-α的表达量无明显变化(P>0.05)。结论大鼠气道内广泛分布有TLR 4和TNF-α, ALI肺组织内TLR 4和TNF-α的表达显著增高,而吸入适当浓度NO可抑制其增高。
Objective To observe the effects of inhalation of nitric oxide(NO) in different concentrations on the degree of pulmonary edema and the expression of toll-like receptor 4(TLR 4) and tumor necrosis factor alpha(TNF-α) in rats' respiratory tract.Methods Thirty-six male SD rats were randomly divided into control group, model group, 10×10~(-6) mg/L NO group, 20×10~(-6) mg/L NO group, 40×10~(-6) mg/L group and 100×10~(-6) mg/L NO group,each group with 6 rats. All groups were given lipopolysaccharide(LPS) to establish acute lung injury(ALI) model of rat except control group.The degree of pulmonary edema and the expression of TLR 4 and TNF-α were observed after inhaled NO of 10×10~(-6) mg/L, 20×10~(-6) mg/L, 40×10~(-6) mg/L, 100×10~(-6) mg/L, respectively.Results The TLR 4 and TNF-α were distributed widely in rats' respiratory tract of control group. Compared with control group, the expression of TLR 4 and TNF-α and the degree of pulmonary edema were significantly increased than those in model group and each NO group(all P<0.05).Compared with model group, the expression of TLR 4 and TNF-α and the degree of pulmonary edema were significantly decreased than those in 10×10~(-6)mg/L NO group, 20×10~(-6)mg/L NO group, 40×10~(-6)mg/L group(all P<0.05), the degree of pulmonary edema were significantly increased in NO 100×10~(-6)mg/L group(P<0.05),but the expression of TLR 4 and TNF-α were no significantly different in NO 100×10~(-6) mg/L group(P>0.05).Conclusion The TLR 4 and TNF-α are distribute widely in rats' respiratory tract,and the expression of TLR 4 and TNF-α increase obviously in ALI of rat, while the increase can be inhibited by inhaling NO at appropriate concentrations.
Objective To observe the effects of inhalation of nitric oxide(NO) in different concentrations on the degree of pulmonary edema and the expression of toll-like receptor 4(TLR 4) and tumor necrosis factor alpha(TNF-α) in rats' respiratory tract.Methods Thirty-six male SD rats were randomly divided into control group, model group, 10×10~(-6) mg/L NO group, 20×10~(-6) mg/L NO group, 40×10~(-6) mg/L group and 100×10~(-6) mg/L NO group,each group with 6 rats. All groups were given lipopolysaccharide(LPS) to establish acute lung injury(ALI) model of rat except control group.The degree of pulmonary edema and the expression of TLR 4 and TNF-α were observed after inhaled NO of 10×10~(-6) mg/L, 20×10~(-6) mg/L, 40×10~(-6) mg/L, 100×10~(-6) mg/L, respectively.Results The TLR 4 and TNF-α were distributed widely in rats' respiratory tract of control group. Compared with control group, the expression of TLR 4 and TNF-α and the degree of pulmonary edema were significantly increased than those in model group and each NO group(all P<0.05).Compared with model group, the expression of TLR 4 and TNF-α and the degree of pulmonary edema were significantly decreased than those in 10×10~(-6)mg/L NO group, 20×10~(-6)mg/L NO group, 40×10~(-6)mg/L group(all P<0.05), the degree of pulmonary edema were significantly increased in NO 100×10~(-6)mg/L group(P<0.05),but the expression of TLR 4 and TNF-α were no significantly different in NO 100×10~(-6) mg/L group(P>0.05).Conclusion The TLR 4 and TNF-α are distribute widely in rats' respiratory tract,and the expression of TLR 4 and TNF-α increase obviously in ALI of rat, while the increase can be inhibited by inhaling NO at appropriate concentrations.
引文
[1] 陈洪雷,潘铁文,徐志飞. 一氧化氮治疗急性呼吸窘迫综合征的临床进展[J]. 创伤外科杂志,2011,13(4):374-376
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[5] 黄丽华,王纯洁,纳仁高娃,等.栀子苷对LPS诱导的RAW264.7巨噬细胞TLR 4-NF-κB通路的影响[J].细胞与分子免疫学杂志,2013,29(10): 1012-1014
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[9] Uhlig S,Yang Y,Waade J,et al. Differential regulation of lung endothelial permeability in vitro and in situ[J]. Cell Physiol Biochem,2014,34(1):11-19
[10] Liu YL,Liu YJ,Liu Y,et al. Hydroxysafflor yellow A ameliorates lipopolysaccharide-induced acute lung injury in mice via modulating toll-like receptor 4 signaling pathways[J]. Int Immunopharmacol,2014,23(2):649-657
[11] 朱蓉,洪永清. 复方丹参注射液对变应性哮喘小鼠脾巨噬细胞TLR2和TLR 4 mRNA表达的研究[J]. 临床肺科杂志,2009,14(7):895-897
[12] 黄继义,刘才文,林建东,等.内毒素急性肺损伤TLR 4-LPS信号传导对NF-κB活性的影响[J].临床肺科杂志,2014,19(2):223-226
[13] Han SH,Mallampalli RK. The acute respiratory distress syndrome from mechanism to translation[J]. J Immunol,2015,194(3):855-860
[14] Tsai YF,Chu TC,Chang WY,et al. 6-Hydroxy-5,7-dimethoxyflavone suppresses the neutrophil respiratory burst via selective PDE4 inhibition to ameliorate acute lung injury[J]. Free Radic Biol Med,2017,106:379-392
[15] 洪小样,孔祥东,周更须. 肺表面活性物质治疗婴幼儿心脏术后急性肺损伤研究[J]. 中华儿科杂志,2012,50(3):193-196
[16] Troncy E, Francoeur M, Blaise G. Inhaled nitric oxide: clinical applications,indications and toxicology[J]. Can J Anaesth, 1997, 44(9): 973-988
[17] Wright CJ,Agboke F,Chen F,et al.NO inhibits hyperoxia-induced NF-κB activation in neonatal pulmonary microvascular endothelial cells[J].Pediatr Res,2010,68(6):484-489
[18] Fike CD,Dikalova A,Slaughter JC,et al.Reactive oxygen species—reducing strategies improve pulmonary arterial responses to nitric oxide in piglets with chronic hypoxia—induced pulmonary hypertension[J].Antioxid Redox Signal,2013,18(14):1727-1738
[2] Chen H,Bai C,Wang X. The value of the lipopolysaccharide-induced acute lung419 injury model in respiratory medicine[J].Expert Rev Respir Med,2010,4(6):773-783
[3] Huang HL,Lin CW,Chiang MF,et al. Lipopolysaccharide directly stimulates aldosterone production via toll-like receptor 2 and toll-like receptor 4 related PI(3)K/Akt pathway in rat adrenal zona glomerulosa cells[J]. J Cell Biochem,2010, 111(4):872-880
[4] Rehberg S, Maybauer MO, Maybauer DM, et al. The role of nitric oxide and reactive nitrogen species in experimental ARDS [J]. Front Biosci(Schol Ed),2010,2(1):18-29
[5] 黄丽华,王纯洁,纳仁高娃,等.栀子苷对LPS诱导的RAW264.7巨噬细胞TLR 4-NF-κB通路的影响[J].细胞与分子免疫学杂志,2013,29(10): 1012-1014
[6] Ma LJ, Li WP, Jin GF. Research progress of pathogenesis of acute lung injury and acute respiratory distress syndrome[J].Chin J Lung Dis,2013,6(1):49-52
[7] Cheng IW, Matthay MA. Acute lung injury and the acute respiratory distress syndrome[J]. Crit Care Clin,2003,19(4):693-712
[8] 付玉梅,陈旭昕,韩志海. Toll 样受体 4 信号传导通路在急性肺损伤发病机制中的作用[J]. 转化医学杂志,2015,4(4):236-239
[9] Uhlig S,Yang Y,Waade J,et al. Differential regulation of lung endothelial permeability in vitro and in situ[J]. Cell Physiol Biochem,2014,34(1):11-19
[10] Liu YL,Liu YJ,Liu Y,et al. Hydroxysafflor yellow A ameliorates lipopolysaccharide-induced acute lung injury in mice via modulating toll-like receptor 4 signaling pathways[J]. Int Immunopharmacol,2014,23(2):649-657
[11] 朱蓉,洪永清. 复方丹参注射液对变应性哮喘小鼠脾巨噬细胞TLR2和TLR 4 mRNA表达的研究[J]. 临床肺科杂志,2009,14(7):895-897
[12] 黄继义,刘才文,林建东,等.内毒素急性肺损伤TLR 4-LPS信号传导对NF-κB活性的影响[J].临床肺科杂志,2014,19(2):223-226
[13] Han SH,Mallampalli RK. The acute respiratory distress syndrome from mechanism to translation[J]. J Immunol,2015,194(3):855-860
[14] Tsai YF,Chu TC,Chang WY,et al. 6-Hydroxy-5,7-dimethoxyflavone suppresses the neutrophil respiratory burst via selective PDE4 inhibition to ameliorate acute lung injury[J]. Free Radic Biol Med,2017,106:379-392
[15] 洪小样,孔祥东,周更须. 肺表面活性物质治疗婴幼儿心脏术后急性肺损伤研究[J]. 中华儿科杂志,2012,50(3):193-196
[16] Troncy E, Francoeur M, Blaise G. Inhaled nitric oxide: clinical applications,indications and toxicology[J]. Can J Anaesth, 1997, 44(9): 973-988
[17] Wright CJ,Agboke F,Chen F,et al.NO inhibits hyperoxia-induced NF-κB activation in neonatal pulmonary microvascular endothelial cells[J].Pediatr Res,2010,68(6):484-489
[18] Fike CD,Dikalova A,Slaughter JC,et al.Reactive oxygen species—reducing strategies improve pulmonary arterial responses to nitric oxide in piglets with chronic hypoxia—induced pulmonary hypertension[J].Antioxid Redox Signal,2013,18(14):1727-1738