| Objective: To establish a scientific and reproducible animal model of sepsis-induced lung injury.To observe the pathological characters and the levels of plasma thrombomodulin (TM), serum tumor necrosis factor-α(TNF-α) and the expression of Nuclear factor-κB (NF-κB) in lung tissues of sepsis rats. To investigate the protective effect of xubijing injection.Methods: In this study, SD rats were subjected to sepsis induced by cecal ligation and puncture (CLP) surgery.One hundred and forty-four normal Sprague-Dawley (SD), weighted 200~230g were randomly divided into four groups: control group (group A, n=24), sham operation group (group B, n=24), CLP group (group C, n=48) and xuebijing-treated group (group D, n=48). Each group was divided into four subgroups. Animals in the subgroups were orderly sacrificed in 6, 24, 48, 72 hours after the operations. Animals in group C were given respectively intravenous injection with xuebijing at the dose of 4ml/kg at post-CLP. Animals in group B and group D were received the same dose of saline. After CLP, rats in each group were anesthetized with 10% chloral hydrate (3ml / kg) at the required time. Blood samples from abdominal artery were collected to measure the levels of TM and TNF-α. Part of the right lung was harvested to observe the pathological characters by Hematoxylin-Eosin (HE) and the expression of NF-κB by immunohistochemistry.Results:①The general situation of rats: The rats in group B were all survived and came around earlier than those in group C and group D. No abdominal dropsy and evil smelling and bowels necroses were found when their abdomens were opened. The rats in group C came around later than those in group B and were observed apparent inaction, piloerection, and tachypnea. Bloody ascites and evil smelling were found when morbid anatomy was performed. Enteric edema and the cecal necrosis were also observed. Compared with the group C, the clinical appearances of the rats in group D were better.②HE staining: The lungs presented pink in both group A and group B. The normal alveolar wall was complete and clear, without soakage of inflammatory cells in the lumen of alveoluses, or occasionally a few of erythrocytes. The lungs were deep red and obviously swollen in group C. There were some ecchymosises in the surface of the lung. At the earlier stage, pulmonary edema, diffuse pulmonary hemorrhage, the infiltration of inflammatory cells and hyaline membrane were observed; At the later stage, part alveolar has collapsed and we also observed bullae. The pathological changes were obvious 24hours after the operation and no imtrpvement could be observed. Compared with group C, rats in group D presented better in pathological changes, but still worse than group B.③TM: The levels of TM in group C gradually increased from 6h to h after CLP surgery. The levels of TM in group C were obviously higher than those in group B on the four time points, there was a statistical significance between them (p<0.01). The levels of TM in group D also gradually increased from 6h to 72h after CLP surgery, but the levels of TM in group D were obviously lower than those in group C, there was a statistical significance between them (p<0.01). The levels of TM in group D were higher than those in group B, there was a statistical significance between group B and group D (p<0.01).④TNF-α: The levels of TNF-αin group C gradually increased from 6h to 72h after CLP surgery and the levels were obviously higher than those in group B and group D, there was a statistical significance between them (p<0.01). The levels of TNF-αin group D also gradually increased from 6h to 72h ,but the levels of TNF-αin group D were obviously lower than those in group C, there was a statistical significance between them (p<0.01). The levels of TNF-αin group D were higher than those in group B, there was a statistical significance between group D and group B (p<0.01).⑤NF-κB expression in lung tissue: There was only a very weak expression of NF-κB in group A and group B. The expression of NF-κB in group C was observed from 6h to 72h in cytoplasma,of bronchial epithelial cells, alveolar epithelial cells and vascular endothelial cells. The expression of NF-κB in group D was observed in 6h in such cells and the expression decreased in 48h and 72h, there was a statistical significance between group C and group D (p<0.05).Conclusion:1 a scientific and reproducible animal model of sepsis-induced lung injury can be established by CLP.2 Xuebijing performs protective effect on animals with acute lung injury after sepsis by decreasing the levels of TNF-αand TM, depressing the expression of NF-κB.
|
PDF Full Text Request