| Background:Acute lung injury(ALI)/ acute respiratory distress syndrome(ARDS)secondary to severe infection,trauma,aspiration,ischemia and reperfusion,acidosis and other reasons,After out of control inflammation of the second hit,the lungs are often the first damaged organs,is one of the most common serious illness in the ICU.Further development can occur serious respiratory failure and life-threatening.Although ALI / ARDS etiology is different,common features are refractory hypoxemia and progressive respiratory distress.The exact pathogenesis is still not fully elucidated.Treatment is also complicated by the original disease,the effect is limited.At present,the main clinical use of small tidal volume of lung protective ventilation strategy,strict liquid control management and CRRT,ECMO and other high-end equipment applications.Although the use of treatment methods and ICU wards in the past two decades has improved the prognosis of the disease,higher incidence and mortality,treatment difficulties,high medical costs and other issues have been plaguing the medical community,as a global Important public health issues.Therefore,new methods of treatment are urgently sought to improve the quo.In recent years,it has been found that stem cells with multi-directional differentiation potential and anti-inflammatory effects are expected to improve these symptoms.Adipose-derived stem cells(ADSCs)are a kind of adult stem cells discovered in recent years,with the advantages of rich source,easy to draw,and the proliferation of strong,low immune function have attracted wide attention.At present,there are few studies on ADSCs were induced by lipopolysaccharide(LPS)in ALI.This study further explored the effect of ADSCs on the expression of inflammatory factors in LPS-induced acute lung injury in rats and its anti-inflammatory mechanism.Objective:Establishment of LPS-induced rat ALI model;labeling of stem cells and tracer stem cells with thymidine analog,5-ethynyl-2-deoxyuridine(EdU)in the lungs;To observe the effect of adipose-derived stem cells(ADSCs)on the therapeutic effect and inflammatory factor of lipopolysaccharide(LPS)-induced acute lung injury(ALI)in rat,and to further explore the mechanism of anti-inflammatory.Methods:70 male SD rats were randomly divided into three groups by the number table way: control group(10 rats),LPS model group(30 rats),LPS+ADSCs group(30 rats);ADSCs were cultured in the 4th generation by adding EdU to incubate the mark stem cells for 24 hours.ALI model was made by intraperitoneal injection of 8 mg/kg LPS,LPS+ADSCs group was received tail vein injection of 300?L ADSCs after 30 minutes for the model was made,the samples of control group were not disposed and extracted immediately,and the remained two groups specimens were taken at 6,24,72 h respectively.Arterial oxygen tension(PaO2)and lactate level from femoral artery.Enzyme-linked immunosorbent assay(ELISA)was used to detected the serum myeloperoxidase(MPO)and interleukin-10(IL-10)from left ventricle blood.Lung wet / dry weight ratio was detected by thoracotomy and the pathological changes of lung tissue were observed under an optical microscope.Observe the location of EdU-labeled stem cells in lung tissue under the fluorescence microscope.Western blot assay was used to detected the protein expression of nuclear factor-?B(NF-?B)from lung tissue of rats.Results:ADSCs labeled with EdU were different degrees of fixation in the lungs at 6 hour,24 hour and 72 hour after tail vein injection.Compared to the control group,the damage degree of lung tissue of LPS group was significantly higher at 6 hour,and lung W/D ratio,blood lactate,MPO,IL-10 and expression level of NF-?B in lung tissue were significantly increased respectively,while PaO2 was decreased significantly.Compared with LPS group,the damage degree of lung tissue of ADSCs intervention group was significantly reduced from 6 hour,and lung W/D ratio,the blood lactate,MPO,and NF-?B expression in lung tissue were significantly decreased,while PaO2 was increased significantly,and it became normal at 72 hour[lung W/D ratio:5.33±0.29 vs 5.77±0.42 at 6 hour,5.14±0.46 vs.5.43±0.38 at 72 hour;blood lactate(mmol/L):3.6±1.0 vs.5.7±1.1 at 6 hour and 3.1±1.0 vs.3.8±1.2 at 72 hour;blood MPO(?g/L):1.50±0.90 vs.2.70±1.85 at 6 hour and 0.46±0.30 vs.0.71±0.22 at 72 h;NF-?B(grayscale value): 0.40±0.11 vs.0.50±0.09 at 6 hour and 0.24±0.03 vs.0.33±0.06;PaO2(mmHg,1 mmHg=0.133 kPa): 78.0±4.1 vs.74.5±3.2 at 6 h and 89.3±9.4 vs.81.9±3.4 at 72 h;P<0.05],The IL-10 level was only higher than the LPS group at 24 hour(ng/L:27.75±15.49 vs.17.52±6.56,P<0.05).Conclusion:ADSCs transplantation can set in LPS-induced ALI rats lung,can improve the oxygenation of PaO2 to reduce serum lactate levels,reduce serum inflammatory factors and lung inflammation,probably by ADSCs inhibiting the expressions of NF-?B. |
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