| Objectives: To study the pathogenesis of sepsis-induced acute lung injury and protective effect of xuebijing injection through establishing rat models of sepsis-induced lung injury by cecal ligation and puncture (CLP) and observing the pathological changes, the expression of NF-κB in lung and the levels of APC and TNF-αin serum of sepsis rats.Methods: 136 normal Sprague-Dawley (SD) adult rats (female and male in half) weighting 200-230g were randomly divided into the following 4 groups. The blank control group (group A, n = 24), Sham operation group(group B, n = 24), CLP group (group C, n = 44), Xuebijing-treated group (group D, n = 44). In this study, sepsis was induced by cecal ligation and puncture (CLP). Each group was divided into 4 subgroups (6, 24, 48, 72 hours) according to 4 different intervals. Animals in group D were given respectively intravenous injection with xuebijing at the dose of 8ml / kg at post-CLP. Animals in group B and group C were received the same dose of saline. Right after CLP, all the adult rats were anesthetized, collected blood sample through the abdominal artery, then measured the TNF-αand APC level. Each rat's inferior lobe of right lung was cut to observe the pathology by Hematoxylin-Eosin (HE) and the expression of NF-κB by immunohistochemistry.Results:1 The general situation of rats in the experiment: The rats in group B analepsised 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 had serious septic performance. They analepsised later, and then were depressed with inaction, piloerection, and tachypnea. When the abdomen was opened, bloody ascites and evil smelling were found, the bowels looked edema and the cecum was necrosed. Compared with the group C, the clinical appearances in the rats of group D were better.2 Pathologic changes: In group A, the lung structure including alveolar septa and alveolar space were normal without edema, congestion or inflammatory cell infliltration. There was no obvious change in group B compared with group A. In group C, the alveolar septum congestion and leakage of fibroprotein could be seen. There were a large number of inflammatory cells including some neutropil, erythrocyte and lymphocyte. Surrounding the vessels and trachea, there were different degree of alveolar septa widen, congestion and pulmonary bulla, which reached peak at 24 hours, and not relieved until the end of this study. The aforementioned changes were also shown up in group D, but were more palliative than that in group C.3 APC: The level of APC in serum decreased in group C at 6 hours post-CLP, indicating a statistical difference compared with group A and B (P < 0.05).The level of APC decreased gradually. In group D, the level of APC was higher than group C at 6 hours post-CLP, indicating a statistical difference between them (P < 0.05).There was no difference between them from 24 to 48 hours (P > 0.05), though the level of APC in group D was still higher than group C. The level of APC in serum in group D was lower than that in group B in all these time points, and there was a statistical difference between them (P < 0.05).4 TNF-α: The level of TNF-αin serum in group C gradually increased from 6 to 72 hours after CLP,and there was a difference compared with group A and B (P < 0.05). The level of TNF-αin serum in group D was lower than that in group C in all these time points,there was a statistical difference between them (P < 0.05). The level of TNF-αin serum in group D was higher than that in group A and B in all these time points, there was a statistical difference between them (P < 0.05).5 NF-κB expression in lung tissue:In group A and B, NF-κB positive cells were found dispersedly by immunohistochemistry; In group C NF-κB expressed increasingly at 6 hours after CLP, reached maximum at 48 hours, lasted to 72 hours. There was a statistical difference between group C and B (P < 0.05). The expression of NF-κB in group D were lower than that in group C in all these time points, and there was a statistical difference between them (P < 0.05). The expression of NF-κB in group D were higher than that in group A and B in all these time points, there was a statistical difference between them (P < 0.05).6 The fatality rate of rats: There was no dead rats in group A and B until 72h. Thirteen rats died in group C (fatality rate 29.55%) and five rats died in group D (fatality rate 11.36%). The fatality rate of group C is higher than group D. There was a statistical difference between group C and D (x2 = 4.47, P < 0.05).Conclusion:1 The sepsis model could be made by cecal ligation and puncture (CLP) successfully.2 The level of serum TNF-αincreased and the level of serum APC decreased in sepsis rats; The expression of NF-κB also increasd. It demonstrated that inflammatory response could inhibit the PC pathway in sepsis.3 Xuebijing could reduce the acute lung injury by decreasing the level of TNF-αand the expression of NF-κB obviously, and increasing the expression of APC. Xuebijing could decrease the fatality rate of spesis rats. |
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