The Role Of Osteopontin On Inflammatory Factors At Acute Lung Injuried Pulmonary Fibrosis Rats Induced By Endotoxin

Objective:For investigating the role of osteopontin and futherpathogenesis in acute lung injuried lung fibrosis,we observe the nuclearfactor kappaB （NF-κB） and IFNlammatory cellscytokines expressionlevels,such as tumor necrosis factor （TNF-α）， interferon gamma（IFN-γ）,Interleukin-4（IL-4）,Interleukin-10（IL-10）in SD rats’lungtissue after intraperitoeally with lipopolysaccharid（eLPS）and osteopontinantibody.Methods:Acute lung injuried lung fibrosis SD rats model wasestablished by intraperitonesl injection with LPS.60femaleSprague-Dawley rats were randomly divided into three groups, controlgroup (n=20,the intraperitoneal injection of sterilesaline the NS),endotoxin model group (n=20,the intraperitoneal injection of LPS,5mg/kg), the intervention group (n=20, the intraperitoneal injection ofLPS,5mg/kg, intraperitoneal injection of osteopontin antibody0.5mltiter1:32after5minutes), Also randomly divided into for groups based ondifferent time points3d,7d,14d,28d after randomly divided into threegroups. three groups were established lung fibrosis model by injectiondrug three times., Every rats in each group were sacrificed on above pointin time,and specimens for corresponding detection.①observed lungtissue pathological changes by HE staining and Masson staining withright lower lung leaves②observed nucleus of NF-κB expression in lung tissue by immunoblot (Western blot) with right upper lung leaves③observed IFNlammatory cytokines expression level such as TNF-alpha,IFN-γ, IL-4, IL-10by enzyme-linked immunosorbent assay (ELLISA)with rats left lung tissue Results:1, The pathology was observed underHE staining and Masson staining200times microscope NS grouprats’lung tissue were normal, no destruction, no IFNlammatory, nopulmonary interval broadened and IFNlammatory cell IFNiltration; whileendotoxin group the alveolar architecture and alveolar septum weredestructed, alveolar spaces closed and integration IFNlammatory cellsfilled the alveolar spaces and pulmonary interstitium, accompaniedbleeding, edema,blue collagen fibers began to deposited in the heavierparts at3d and7d groups;Later,the inflammatory response weaken butmore blue collagen fibersthe appeared at injuried site of the lung.Compared with endotoxin group the intervention group pathologicalmanifestations were weaken during the whole experiment, comparedwith the NS group endotoxin group alveolitis score was significantlyincreased (p<0.01, p<0.05) the peak period of inflammation in the3d,7d,while the intervention group was significantly more decreased (p<0.01)compared with NS group, the difference was not statistically significant(p>0.05) at28days Endotoxin group and intervention group, lungfibrosis score increased (p<0.05, p<0.01) than those in NS group but thedegree of pulmonary fibrosis in intervention group was lower than endotoxin group (p<0.05,p<0.01)2,compared with NS group thecytoplasm and nucleus of NF-κB expression in rat lung tissuehomogenates began to rise in the early time,which was statisticallysignificant (p<0.01) and the3days7days groups were most obviousbetween model group and intervention group.cytoplasm of NF-κBexpression, including no difference between the toxin model group andintervention group (p>0.05), while the nucleus of NF-κB expressionbetween the two groups which was statistically significant (p<0.01).3,The proIFNlammatory cytokines expression level such as TNF-alpha,IFN-γ increased at an early stage and the amount existed consistency withthe nucleus of NF-kappa B still8h and7d groups is the most obviousbetween endotoxin model group and intervention group in lunghomogenates At the same time the two groups compared with NS groupthere was significant difference (p<0.05, p<0.01), later,theIFNlammatory factor expression gradually decreased, the quantity ofproIFNlammatory cytokines TNF-alpha and IFN-γ expression in theintervention group creased significantly than model group(p<0.05,p<0.01); while anti-IFNlammatory cytokines IL-4, IL-10in the modelgroup and intervention group remained significantly higher (p<0.05,p<0.01),compared with NS group also intervention group increased moresignificantly (p<0.05)compared with model group.Conclusion:multiplesmall intraperitoneal injection with LPS can copy acute lung injury and lung fibrosis model, the most serious injury period is in8h and7d, themost significant fibrosis moment is in28d. The expression of NF-κB andIFNlammation of TNF-alpha, IFN-γ, IL-4, IL-10increased in the earlytime in endotoxin model of lung tissue and the peak is3d，7d after theexpression level was gradually decline over time pulmonary fibrosisassociated with NF-κB and inflammation of TNF-alpha and IFN-γ, IL-4,IL-10Osteopontin involved in the process of pulmonary fibrosis byactiving NF-κB pathway, increasing of proinflammatorycytokines,decreasing the anti-inflammatory factor.The use of a certaindose of osteopontin antibody can weaken this fibrosis at certain extent.