摘要
目的:探讨重度吸入性损伤后气管切开患者气道呼出气冷凝液(EBC)中多种细胞因子水平变化及其临床意义。方法:采用前瞻性研究方法,选择2021年5月至2022年8月南京医科大学附属苏州医院烧伤整形科收治的32例烧伤合并重度吸入性损伤患者;选择同期20例健康志愿者作为对照。收集患者伤后12 h EBC,同时收集健康对照者样本,采用液相芯片技术测定EBC中27种细胞因子水平,其中包括肿瘤坏死因子-α(TNF-α)和白细胞介素(IL-1β、IL-6、IL-8、IL-10、IL-17)6种炎症细胞因子;收集患者伤后12 h血浆,同时收集健康对照者血浆,采用液相芯片技术检测上述6种炎症细胞因子水平,分析其与EBC中含量的差异;于患者伤后12 h及3、7、14、21 d收集血浆和EBC,采用酶联免疫吸附试验(ELISA)检测TNF-α水平。结果:最终32例患者纳入分析,烧伤总面积(40±16)%总体表面积(TBSA);入院时间为伤后(4.2±2.3)h。①EBC中27种细胞因子:重度吸入性损伤患者巨噬细胞炎症蛋白-1β(MIP-1β)、IL-6、IL-5、IL-2、IL-1β、IL-8、IL-10、IL-15、IL-9、γ-干扰素(IFN-γ)、IL-1受体拮抗剂(IL-1ra)、TNF-α、嗜酸粒细胞趋化因子(Eotaxin)、碱性成纤维细胞生长因子(bFGF)、血小板衍生生长因子-BB(PDGF-BB)、干扰素诱导蛋白-10(IP-10)、巨噬细胞趋化蛋白-1(MCP-1)、粒细胞集落刺激因子(G-CSF)18种细胞因子水平均较健康对照者明显升高,其中Eotaxin在健康对照者EBC中未检出;粒-巨噬细胞集落刺激因子(GM-CSF)、趋化因子配体5(CCL5/RANTES)、IL-13、IL-4、MIP-1α5种细胞因子在重度吸入性损伤及健康对照者EBC中均未检出;重度吸入性损伤患者EBC中血管内皮生长因子(VEGF)和IL-12 p70较健康对照者轻度下降,IL-7和IL-17轻度升高,但差异均无统计学意义。②血浆中6种炎症细胞因子:重度吸入性损伤患者IL-6和IL-8水平均较健康对照者明显升高〔IL-6(ng/L):18.51(10.87,26.21)比0.22(0.10,0.36),IL-8(ng/L):10.75(8.58,18.79)比1.06(0.81,2.14),均P<0.01〕;TNF-α、IL-1β、IL-10在重度吸入性损伤患者血浆中轻度升高,IL-17轻度下降,但与健康对照组比较差异无统计学意义。而同期采集的EBC中,重度吸入性损伤患者TNF-α、IL-1β、IL-6、IL-8、IL-10这5种炎症细胞因子均较健康对照者明显升高〔TNF-α(ng/L):16.42(12.57,19.21)比7.34(6.11,8.69),IL-1β(ng/L):15.57(10.53,20.25)比0.99(0.67,1.41),IL-6(ng/L):13.36(9.76,16.54)比0.70(0.42,0.85),IL-8(ng/L):1059.29(906.91,1462.37)比10.36(8.40,12.37),IL-10(ng/L):2.69(1.54,3.33)比1.54(1.18,2.06),均P<0.05〕。③血浆和EBC中TNF-α动态变化:重度吸入性损伤患者EBC中TNF-α水平整体低于血浆。血浆TNF-α水平随伤后时间延长逐渐升高,3 d时明显高于健康对照者〔ng/L:30.38(24.32,39.19)比22.94(17.15,30.74),P<0.05〕,14 d达到高峰,随后回落;而EBC中TNF-α水平于伤后12 h即较健康对照者明显升高〔ng/L:15.34(11.75,18.14)比6.99(6.53,7.84),P<0.01〕,3 d即达到峰值,随后逐渐回落。结论:重度吸入性损伤患者EBC中存在多种细胞因子表达变化,其中包括TNF-α在内的多种炎症细胞因子变化较血浆中的变化更敏感,可以应用于吸入性损伤的病情监测评估。
Objective To investigate the changes and clinical significance of multiple cytokine levels in exhaled breath condensate(EBC)in patients undergoing tracheotomy with severe inhalation injury.Methods A prospective study was conducted.A total of 32 patients with severe burn combined with severe inhalation injury admitted to the department of burns and plastic surgery of Affiliated Suzhou Hospital of Nanjing Medical University from May 2021 to August 2022 were enrolled.Twenty healthy volunteers from the same period were served as controls.EBC of patients at 12 hours after burn and the samples of healthy controls were collected.The levels of 27 cytokines in EBC,including tumor necrosis factor-α(TNF-α)and interleukins(IL-1β,IL-6,IL-8,IL-10,and IL-17),were determined by liquid phase chip technology.Meanwhile,plasma of patients at 12 hours after burn and the plasma of volunteers were collected,and the levels of inflammatory cytokines were detected by liquid chip technology,and the differences between the levels in plasma and those in EBC were analyzed.Plasma and EBC of patients with aspiration injury were collected at 12 hours and 3,7,14 and 21 days after burn,and TNF-αlevels were determined by enzyme-linked immunosorbent assay(ELISA).Results Finally,32 patients were enrolled,and the total burned area was(40±16)%of total body surface area(TBSA).The time of admission was(4.2±2.3)hours after injury.①Twenty-seven cytokines in EBC:18 kinds of cytokines including macrophage inflammatory protein-1β(MIP-1β),IL-6,IL-5,IL-2,IL-1β,IL-8,IL-10,IL-15,IL-9,interferon-γ(IFN-γ),IL-1 receptor antagonist(IL-1ra),TNF-α,chemotactic factor for eosinophil(Eotaxin),basic fibroblast growth factor(bFGF),platelet derived growth factor-BB(PDGF-BB),interferon-inducible protein-10(IP-10),monocyte chemoattractant protein-1(MCP-1),granulocyte colony-stimulating factor(G-CSF)were significantly increased in patients with severe aspiration injury compared with health controls.Eotaxin was not detected in EBC of healthy controls.Five cytokines,granulocyte-macrophage colony-stimulating factor(GM-CSF),chemokine ligand 5(CCL5/RANTES),IL-13,IL-4 and MIP-1α,were not detected in EBC of severe inhalation injury patients and healthy controls.Vascular endothelial growth factor(VEGF)and IL-12 p70 in EBC of severe aspiration injury patients were slightly decreased as compared with healthy controls,while IL-7 and IL-17 were slightly increased,but the differences were not statistically significant.②Six inflammatory cytokines in plasma:the levels of IL-6 and IL-8 in the severe aspiration injury group were significantly increased as compared with healthy controls[IL-6(ng/L):18.51(10.87,26.21)vs.0.22(0.10,0.36),IL-8(ng/L):10.75(8.58,18.79)vs.1.06(0.81,2.14),both P<0.01].The plasma levels of TNF-α,IL-1βand IL-10 were slightly increased in patients with severe aspiration injury as compared with healthy controls,and IL-17 was slightly decreased,but the difference was not statistically significant.In the EBC collected during the same period,five inflammatory cytokines,including TNF-α,IL-1β,IL-6,IL-8 and IL-10,in patients with severe inhalation injury were significantly increased as compared with healthy controls[TNF-α(ng/L):16.42(12.57,19.21)vs.7.34(6.11,8.69),IL-1β(ng/L):15.57(10.53,20.25)vs.0.99(0.67,1.41),IL-6(ng/L):13.36(9.76,16.54)vs.0.70(0.42,0.85),IL-8(ng/L):1059.29(906.91,1462.37)vs.10.36(8.40,12.37),IL-10(ng/L):2.69(1.54,3.33)vs.1.54(1.18,2.06),all P<0.05].③Dynamic changes of TNF-αin plasma and EBC:the level of TNF-αin EBC of patients with severe aspiration injury was lower than that in plasma.Plasma TNF-αlevel was increased gradually with the extension of time after injury,and was significantly higher than that of healthy controls on day 3[ng/L:30.38(24.32,39.19)vs.22.94(17.15,30.74),P<0.05],and reached the peak on day 14,then fell back.The level of TNF-αin EBC at 12 hours after injury was significantly higher than that in healthy controls[ng/L:15.34(11.75,18.14)vs.6.99(6.53,7.84),P<0.01],and reached the peak on 3 days after injury,and then gradually decreased.Conclusion There are changes in the expression of multiple cytokines in EBC of patients with severe inhalation injury,and the changes of many inflammatory cytokines including TNF-αare more sensitive than those in plasma,which can be used to monitor and evaluate the condition of patients with inhalation injury.
作者
刘云峰
张勇
丁盛
孙燃
刘小俊
苏建东
孙炳伟
Liu Yunfeng;Zhang Yong;Ding Sheng;Sun Ran;Liu Xiaojun;Su Jiandong;Sun Bingwei(Department of Burns and Plastic Surgery,Affiliated Suzhou Hospital of Nanjing Medical University(Suzhou Municipal Hospital),Suzhou 215002,Jiangsu,China)
出处
《中华危重病急救医学》
CAS
CSCD
北大核心
2023年第8期818-822,共5页
Chinese Critical Care Medicine
基金
国家自然科学基金(U21A20370)
江苏省苏州市科技计划项目(SKJY2021117)。
