白藜芦醇对内毒素性急性肺损伤小鼠核转录因子-κB及炎症反应的影响

Effects of resveratrol on changes of nuclear factor-K B and inflammatory reaction in mice with acute lung injury induced by lipopolysaccharide

目的研究白藜芦醇对内毒素脂多糖（I．Ps）诱导的急性肺损伤（ALl）小鼠核转录因子-KB（NF—KB）及炎症反应的影响。方法雄性ICR小鼠32只，按随机数字表法分为对照组、模型组、白藜芦醇预处理组、地塞米松阳性对照组，每组8只。白藜芦醇预处理组以白藜芦醇25mg／kg灌胃，地塞米松阳性对照组腹腔注射地塞米松10mg／kg，对照组、模型组灌胃1ml生理盐水，共3d。第4日尾静脉注射LPS15mg／kg，对照组注射等量生理盐水，8h后处死小鼠。检测肺组织湿／干质量（W／D）比值、苏木素-伊红（HE）染色观察肺组织病理学变化，蛋白质免疫印迹法（Westernblotting）检测NF—KBp65蛋白表达，实时荧光定量聚合酶链反应（PCR）检测白细胞介素-1B（IL—l13）、巨噬细胞炎性蛋白-1d（MIP-1d）的mRNA表达。结果与对照组比较，模型组小鼠肺损伤明显，肺泡内出血、水肿、炎性细胞浸润，肺组织w，D比值增加，NF—KB活化表达明显增加，肺组织IL-113和MIP-1dmRNA表达均明显增加（均｜P〈0．01）。与模型组比较，白藜芦醇预处理组、地塞米松阳性对照组肺组织病理损伤程度明显减轻，肺组织W／D比值（5．23±0．15、5．21±0．21比5．48±0．14）、NF—KBp65赁自（1．25±0．09、1．09±0．04比1．52±0．10），IL-1BmRNA（11．10±2．84、5．10±0．82比23．00±4．25）和MIP-1dmRNA（29．01±5．8l、12．30±0．95比60．27±5．62）相对表达量均明显降低（均P〈0．01）。结论白藜芦醇对内毒素性ALI有显著的保护作用，其作用机制可能与其抑制NF—KB活化、降低肺组织炎症反应有关。

Objective To investigate the effect of resveratro] on changes of nuclear faetor-KB （NF-KB） and inflammatory reaction in miee with acute lung injury （ALl） induced by lipopolysaccharide （LPS）. Methods According to random number table method,32 male ICR mice were divided into four groups： control, model, resveratrol pretreatment and dexamethasone positive control groups （each, n=8）. The mice in the resveratrol pretreatment group were orally gavaged with 25 mg/kg resveratrol for 3 days before injection with LPS, ｜0 mg/kg of dexamethasone was injected intra-peritoneally for 3 days in the positive control group, and 1 ml of normal saline was gavaged for 3 days ill the control and model groups. On the 4th day, LPS 15 mg/kg was injected via a tail vein in the above three groups, while in the control group, the same amount of normal saline was injected into the vein, and after 8 hours, the mice were sacrificed. The lung wet/dry weight （W/D） ratio and hematoxylin-eosin （HE） staining for investigation of the histopathologie changes of lung tissues were carried out. The Western blotting was used to detect the protein expression of NF-KB p65, real-time fluoreseein quantitative polymerase chain reaction （PCR）was applied to assay the expression of interleukin-1 [3 （IL-1 [3 ） mRNA and macrophage inflammatory protein-1 c~ （MIP-1 e~ ） mRNA in lung tissue. Results Compared with the control group, in the model group, there was significant lung structural damage, the histological results showed pulmonary alveolar hemorrhage, edema and inflammatory cell infiltration ; the lung W/D ratio, the activity and its expression of NF-KB, the expressions of IL-I ]3 and MlP-1 α mRNA in lung tissue were significantly increased （all P〈 0.01 ） . Compared with the model group, the lung histopathological changes were much more ameliorated, and the W/D ratio （5.23_＋0.15,5.21 _＋0.21 vs. 5.48_＋0.14）, the activity of NF-xB p65 protein （1.25_＋0.09, 1.09_＋0.04 vs. 1.52_＋0.10）, the mRNA expressions of IL-113 （11.10_＋2.84,5.10_＋0.82 vs. 23.00_＋4.25）and MIP-1 cL （29.01 _＋ 5.81,12.30_＋ 0.95 vs. 60.27 ＋ 5.62） were markedly suppressed in resveratrol pretreatment group and dexamethasone positive control group （all P〈0.01）. Conclusion Resveratrol has remarkable protective effect on ALl induced by LPS in mice, its mechanism is possibly related to the inhibition of NF-KB activation and reduction of inflammatory response in the lung tissue.