The Study Of The Anti-CD14 Monoclonal Antibody On SICAM-1 And KC Expression Of Lung Tissue With Sepsis In Rat

Objective:To explore protection mechanism of the Anti-CD14 McAb on the treatment of sepsis with acute lung injury in rat.Methods:18 male Wistar rats(200-250)g were randomly divided into 3 groups,group Sham,group CLP and group Anti-CD14.There were 6 rats in each group.The cecal ligation and puncture(CLP)method was used to make the animal model with acute lung injury,group Sham as control group,group Sham rats just flip the cecum without ligation and puncture,the group Anti-CD14 was injected 0.2ml Anti-CD14 monoclonal antibody(1ug/ml)by femoral vein after postoperative,group Sham?CLP were injected saline 0.2ml by femoral vein after postoperative,After the success of modeling 6h,we draw blood from abdominal aorta and detected plasma KC,sICAM-1 of 3 group rats.Inferior lobe of right lung was taken for HE staining and observed the pathological changes in the structure of lung tissue.Sham group as control group.In vitro part: a part of cultivated NR8383 were seeded in 24-well culture plate,divided into three groups,control group(group?),sepsis model group(group?)and Anti-CD14 intervention group(group?).group?was added to the normal plasma,group?was add to the sepsis plasma,group?was added to the sepsis plasma and Anti-CD14 monoclonal antibody,three group was incubated about 1 hour in incubator with 37 ? and 5% CO2,Western blot was used to detect the NF-kappa B(p65)protein expression levels of the NR8383 scraped from24-well culture plate;another part of cultivated NR8383 were seeded in 24-well culture plate with appropriate size of the sterilized cover glass,grouping and intervention measureswas the same as the above,the confocal laser scanning microscope(CLSM)was used to detect the NF-kappa B(p65)protein into the NR8383 nuclear situation.Results:In vivo experiments:Compared with the group Sham[KC(pg/ml):72.645±19.860,sICAM-1(pg/ml):252.766±19.921],KC?sICAM-1 expression level in plasma of the group CLP[KC(pg/ml):490.316±43.403,sICAM-1(pg/ml):686.952±36.411] and the group Anti-CD14[KC(pg/ml):348.150±20.924,sICAM-1(pg/ml):411.050±47.170] significantly increased,but sICAM-1?KC expression levels in plasma of the group Anti-CD14 below the group CLP,there were significant differences(P<0.05);lung tissue pathology results indicate pulmonary interstitial of the group CLP obvious thickening,a large number of inflammatory cells infiltrated the pulmonary interstitial and alveolar cavities,the group Anti-CD14 symptoms significantly lower than the group CLP,but higher than the group Sham;Cell experiment Western blot test shows:the group ?(2.1903±0.1199)and the group ?(1.3658±0.1018)NR8383 NF-kappa B(p65)into the NR8383 nucleus higher than group ?(1.0122±0.0780),the group ?NF-kappa B(p65)into the NR8383 nucleus level lower than the group ?,there were significant differences(P<0.05).Cell experimental CLSM test results indicates:compared with group?(0.131±0.009),the group ?(0.685±0.037)and the group ?(0.210±0.008)NF-kappa B(p65)protein into the NR8383 nucleus increased significantly,while the group ? NF-kappa B(p65)protein into the NR8383 nucleus lower than the group ?,there were significant differences(P < 0.05).Conclusion:Anti-CD14 McAb may control the NR8383 NF-kappa B(p65)protein into the nucleus and reduce its Chemokines KC expression,thus reducing neutrophilic granulocyte infiltration,as a result,Anti-CD14 McAb plays a protective role in sepsis with acute lung injury;with Anti-CD14 McAb inhibit endothelial cells expressing sICAM-1 and reduce neutrophil adhesion function of endothelial cells,therefore,Anti-CD14 McAb plays a lung protective role.