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目的建立急性兔肺挫伤模型,评价基质金属蛋白酶抑制剂(tissue inhibitor of metalloproteinases,TIMPs)对肺组织水通道蛋白1(aquaporin,AQP1)和核因子κB(nuclear factor kappa B,NF-κB)的影响。方法通过撞击模式制备急性兔肺挫伤模型,分为3组(正常组、模型组和TIMP组),每组10只,其中TIMP组采用TIMPs(GM6001)进行干预,24 h后收集标本。免疫组化法检测肺组织中AQP1蛋白表达并计算灰度值;双抗夹心酶联免疫吸附法(enzyme linked immunosorbent assay,ELISA)检测NF-κB的表达水平;苏木精-伊红(hematoxtylin-eosin,HE)染色评价各组肺组织的形态学改变。结果采用GM6001干预24 h后,TIMP组肺组织的湿重/干重比值(wet weight/dry weight,W/D)和NF-κB表达水平明显低于模型组,而AQP1蛋白的表达水平在TIMP组明显高于模型组,且差异具有统计学意义(P<0.05)。HE染色提示TIMP组水肿和炎症反应较模型组有明显改善。结论 TIMP能降低肺组织中NF-κB的表达,上调AQP1蛋白表达,改善肺挫伤引起的炎症反应和肺水肿。
Objective To establish an acute rabbit model of pulmonary contusion and evaluate the effect of tissue inhibitor of metalloproteinases (TIMPs) on the expression of aquaporin (AQP1) and nuclear factor kappa B (NF-κB) . Methods Acute rabbit model of pulmonary contusion was prepared by means of impact mode and divided into 3 groups (normal group, model group and TIMP group) with 10 rats in each group. TIMPs (GM6001) were used in intervention group. The expression of AQP1 protein in lung tissue was detected by immunohistochemistry and the gray value was calculated. The expression of NF-κB was detected by enzyme linked immunosorbent assay (ELISA). The hematoxylin- eosin, HE) staining to evaluate the morphological changes of lung tissue in each group. Results After treated with GM6001 for 24 h, the expression of wet weight / dry weight (W / D) and NF-κB in lung tissue of TIMP group was significantly lower than that of model group, while the expression of AQP1 protein in TIMP Group was significantly higher than the model group, and the difference was statistically significant (P <0.05). HE staining showed TIMP edema and inflammatory response than the model group was significantly improved. Conclusion TIMP can reduce the expression of NF-κB in lung tissue, up-regulate the expression of AQP1 protein and improve the inflammatory reaction and pulmonary edema induced by pulmonary contusion.