| Part I Relationship between inflammatory factors,Th1 / Th2 cell differences and disease severity in patients with sepsis related acute lung injuryObjective:To observe the relationship between inflammatory factors,Th1/Th2 cells and disease severity in patients with different degrees of lung injuryMethods:1.Analysis 388 patients with sepsis treated in our hospital from January2010 to April 2021 were analyzed.The patients with concomitant lung injury were included in the lung injury group(n = 68),and the other patients were included in the non lung injury group(n=320).White blood cell(WBC),neutrophil granulocyte(n),procalcitonin(PCT),C reactive protein(CRP)and Interleukin-8,interleukin-6(IL-8),interleukin-6(IL-6),interleukin-α(Tumor necrosis factor-α,TNF-α)、 The levels of Th1 cells,Th2 cells and Th1/Th2 ratio were determined to determine the differences of various inflammatory factors,serological indexes,Th1,Th2 cells and Th1/Th2 between the two groups.2.Sepsis complicated with lung injury was divided into mild group,moderate group and severe group.The grouping was defined according to the score of Murray lung injury scale.Pearson correlation coefficient was used to calculate the correlation between inflammatory factors,Th1/Th2 cytokines and disease severity in patients with sepsis and lung injury.The ROC curve was used to determine the diagnostic efficacy of each index for lung injury and disease severity.Results:1.There was no significant difference in gender,age and primary disease between acute lung injury group and non injury group(P> 0.05).2.The level of N in acute lung injury group was significantly higher than that in non injury group(P < 0.05);There was significant difference in WBC and N between patients with acute lung injury of different severity(P < 0.05);There were significant differences in PCT,CRP and hs CRP between acute lung injury group and non injury group(P < 0.05).3.The levels of IL-6,IL-8 and TNF-α in acute lung injury group were higher than those in non acute injury group,The levels of Th1 cells and Th1 /Th2 ratio increased significantly(P < 0.05);Comparison of IL-6,IL-8 and TNF-α in patients with acute lung injury of different severity,There were significant differences in Th1 cells,Th2 cells and Th1 / Th2 ratio(P < 0.05).4.For the diagnosis of acute lung injury,hs-CRP > IL-6 > IL-8 > CRP >TNF-α> Th1 cells > N> PCT > Th1 / Th2 ratio.In terms of specificity,IL-8 > hs-CRP > PCT > Th1 / Th2 ratio > IL-6 > Th2 cells > CRP > Th1 cells > N > TNF-α。For the diagnosis of moderate and severe acute lung injury,the sensitivity of each index is more than 95%;In terms of specificity,IL-8 > hs-CRP > Th2 cells > IL-6 > PCT > CRP > Th1 cells > Th1 / Th2 ratio.For the diagnosis of severe acute lung injury,the sensitivity was > 95% except CRP;In terms of specificity,IL-8 > hs-CRP > IL-6 > PCT > CRP > Th1 / Th2 ratio > Th1 cells > n > Th2 cells > TNF-α。Conclusions:1.Patients with sepsis with acute lung injury will be caused by N,PCT,CRP,hs-CRP,IL-6,IL-8 and TNF-α、The increase of Th1 cells and Th1 / Th2 ratio increased the risk of lung injury;The severity of lung injury will increase with N,PCT,CRP,hs CRP,IL-6,IL-8 and TNF-α、Th1 and Th1/Th2 ratio increased,but increased with the decrease of Th2 percentage.2.Patients with no acute lung injury,mild acute lung injury,moderate acute lung injury and severe acute lung injury can use a variety of inflammatory factors,especially hs-CRP,which can be used as an important factor in the evaluation of disease severity.However,for patients with moderate and severe lung injury,IL-6 and IL-8 can be further used as differential indicators,and Th2 cells can form screening value for patients with moderate lung injury;PCT and Th1/Th2 ratio can improve the screening value of patients with severe lung injury.Part II Protective effect and mechanism of high concentration hydrogen on acute lung injury induced by lipopolysaccharide in miceObjective: The purpose of this study is to observe the effects of 66.7%high concentration hydrogen inhalation on pathological reaction and vascular permeability of acute lung injury induced by lipopolysaccharide(LPS),and to explore the role of sirt1/NF-κB/catalase signaling pathway in this process.Methods: Animals or cells were divided into control group,LPS group,LPS+66.7% hydrogen inhalation group and LPS+ resveratrol group.(1)66.7%hydrogen was produced by electrolyzed water.HE staining was used to evaluate the effects of 66.7% hydrogen inhalation and positive control drug resveratrol on lung pathological reaction induced by LPS and the lung injury score was executed.(2)The wet/dry weight ratio of lung tissue was used to detect the degree of pulmonary edema in each group;(3)The levels of IL-1β,TNF-α,IL-8,ICAM-1,VCAM-1 and sirt1 in alveolar lavage fluid of each group were detected by ELISA and PCR,and the levels of MDA in lung tissue of each group were detected by MDA kit;(4)Evans blue dye was used to detect the leakage of pulmonary vascular in each group,and monolayer cell transmembrane resistance was used to evaluate the cell permeability in each group;(5)Immunohistochemistry and western blot were used to detect the effects of 66.7% hydrogen inhalation on the expression of sirt1,NF-κB and catalase in lung tissue.(6)Immunofluorescence was used to detect the influence of 66.7% hydrogen inhalation on nuclear translocation of NF-κB.Results:1.High concentration hydrogen inhalation can reduce the inflammatory response of LPS induced acute lung injury in mice and improve pulmonary edema;It can reduce the inflammatory factor IL-1β、TNF-α、IL-8 in alveolar lavage fluid,decreased the expression levels of ICAM-1 and VCAM-1,and decreased pulmonary oxidative stress response;It can improve the increase of pulmonary vascular permeability induced by LPS and reduce the permeability of pulmonary microvascular endothelial cells.2.The high expression of sirt1 in lung can significantly reduce the release of inflammatory factors induced by LPS;3.The expression of sirt1 decreased significantly in LPS acute lung injury group,while high concentration hydrogen inhalation could significantly increase the expression of sirt1 and reduce NF-κB expression,reduced NF-κB in nucleus.In addition,high concentration hydrogen inhalation can significantly increase the expression of lung antioxidant protein catalase.Conclusion:1.66.7% high concentration hydrogen produced by electrolyzed water has significantly protective effect on lung pathological reaction in acute lung injury caused by LPS;2.66.7% high concentration hydrogen exposure can improve pulmonary vascular permeability and reduce pulmonary edema;3.The improvement of acute lung injury by 66.7% high concentration hydrogen is referred to the increase of sirt1 expression,following reducing the inflammation and oxidative stress in lung. |
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