IL-35 Regulates VCAM-1 In ARDS With Pulmonary Hypertension

Acute Respiratory Distress Syndrome(ARDS)is a common disease in acute and severe diseases.It is caused by pulmonary extra-pulmonary diseases such as infection,shock,trauma,and increases in pulmonary capillary permeability and diffuse injury.Based on this,pulmonary arterial hypertension,pulmonary edema,atelectasis and hyaline membrane formation are the main pathological changes,and progressive respiratory distress and refractory hypoxemia are clinically characterized syndromes.ARDS has a high incidence and mortality,leading to a large number of causes of ARDS,and sepsis is the main cause.Pulmonary hypertension(PH)is one of the basic features of ARDS.Its pathological features include pulmonary vasoconstriction,extensive capillary occlusion and pulmonary artery remodeling.Endothelial dysfunction is an important part of the development of pulmonary hypertension in ARDS.Endothelial cells prevent the adhesion of platelets and leukocytes during organ perfusion and release vasoactive substances to regulate vascular tone.IL-35 is a novel member of the IL-12 cytokine family and is composed of a covalently linked disulfide bond between the IL-12 alpha chain p35 and the IL-27 beta chain Ebi3.IL-35 is a new anti-inflammatory factor that occurs after transforming growth factor-beta(TGF-beta),IL-10,and IL-27,and in many diseases such as chronic hepatitis,collagen-induced arthritis,and autoimmune release.Myelitis and so on play a role in the immune regulation of inflammation,IL-35 is becoming a new target for the treatment of infectious diseases,autoimmune diseases and tumors.IL-35 can inhibit the accumulation of acidic granulocytes and neutrophils,let us consider whether it also plays a similar role in ARDS with pulmonary hypertension can reduce cell aggregation.So we designed the following study.The main contents of this study: 1.Pulmonary arterial pressure measured by sepsis-induced ARDS rat model,detection of IL-35 and s VCAM-1 expression in rat peripheral blood,to explore the merger of IL-35 and s VCAM-1 in ARDS Pulmonary arterial hypertension is related to further research.2.Human pulmonary artery endothelial cells(HPAECs)were cultured in vitro and divided into blank control group,TNF-α stimulation group,and IL-35+TNF-α stimulation group to detect the expression of NF-κB and VCAM-1,and to investigate the expression of HPAECs in HPAECs.Activation of VCAM-1 by IL-35.Part Ⅰ: Expression of IL-35 in the peripheral blood of ARDS rats induced by sepsisObjective:To investigate the expression of IL-35 in the peripheral blood of ARDS rats induced by sepsis.Methods:The sepsis rat model was established by cecal ligation and puncture(CLP)and a sham-operated group was established.Peripheral blood was collected at the end of the tail after 24 hours for hemodynamic detection.Lung pathology confirmed the success of the model,through the liquid phase.The expression of IL-35 and s VCAM-1 in peripheral blood was measured by microarray.The concentrations of IL-6,IL-18,TNF-α,and IFN-γ in alveolar lavage fluid were measured by Elisa method.Results:The concentration of inflammatory cytokines in CLP group was significantly higher than that in Sham group(P<0.05).The concentration of IL-35 in CLP group was significantly lower than that in Sham group(P<0.0001)and the concentration of s VCAM-1 was significantly increased(P<0.0001).And there was a negative correlation between IL-35 and s VCAM-1(r=-0.7803 P<0.001).Pathological sections showed that ARDS occurred.Hemodynamic measurements confirmed the presence of pulmonary hypertension.Conclusions:The decrease of IL-35 concentration and increase of sVCAM-1 concentration were negatively correlated,suggesting that the imbalance between the two may play an important role in the development of ARDS with pulmonary hypertension.Part Ⅱ: The activation of VCAM-1 by IL-35 in HPAECsObjective: To study the activation of VCAM-1 by IL-35 in HPAECsMethods: Human pulmonary artery endothelial cells(HPAECs)were cultured in vitro and divided into blank control group,tumor necrosis factor(TNF-α)stimulation group and IL-35+ TNF-α stimulation group.VCAM-1 was detected by flow cytometry and Western blot.NF-κB P65 protein concentration,fluorescence microscopic observation of adhesion of mononuclear cells and endothelial cellsResults:Compared with TNF-α stimulation group,the expression of VCAM-1 and NF-κB P65 in the endothelial cells stimulated by IL-35+TNF-α decreased(P<0.05),and the adhesion of mononuclear cells to endothelial cells decreased significantly.Conclusions: IL-35 inhibits VCAM-1 activation in HPAECs and protects endothelial cells.