宣白承气汤调控sirt1/FOXO1通路对脂多糖致大鼠急性肺损伤的影响

Effect of Xuanbai Chengqi Decoction Regulating Sirt1/FOXO1 Pathway for Lipopolysaccharide-Induced Acute Lung Injury in Rats

目的:探讨宣白承气汤对脂多糖(LPS)诱导的急性肺损伤大鼠的影响以及可能机制。方法:清洁级SD雄性大鼠50只,尾静脉注射LPS制备急性肺损伤模型,随机分为模型组、阳性对照组(地塞米松)、低剂量宣白承气汤组(2g生药/kg)、中剂量宣白承气汤组(5g生药/kg)、高剂量宣白承气汤组(8g生药/kg),每组10只,另设正常对照组10只,模型制备成功后连续7d给药,末次给药后通过HE染色法观察肺组织病理变化,通过考马斯亮蓝法检测大鼠肺泡灌洗液蛋白含量,通过酶联免疫吸附法检测血清肿瘤坏死因子α(TNF-α)、一氧化氮(NO)水平,采用免疫印迹法(WB)检测大鼠肺组织sirt1/FOXO1通路相关蛋白表达。结果:模型组大鼠肺泡腔内充满炎性渗出物和出血,低、中、高宣白承气汤组炎症细胞浸润及出血情况有所改善;与正常组相比,模型组大鼠肺泡灌洗液蛋白、肺体指数、血清TNF-α、NO水平、大鼠肺组织FOXO1蛋白水平显著升高(P<0.05),大鼠肺组织sirt1蛋白水平显著下降(P<0.05);与模型组相比,低、中、高剂量宣白承气汤组肺泡灌洗液蛋白、肺体指数、血清TNF-α、NO水平、大鼠肺组织FOXO1蛋白水平均依次降低(P均<0.05),大鼠肺组织sirt1蛋白水平依次升高(P<0.05),且高剂量宣白承气汤组肺泡灌洗液蛋白、肺体指数、血清TNF-α、NO水平、大鼠肺组织FOXO1、sirt1蛋白水平与阳性对照组差异无统计学意义(P>0.05)。结论:宣白承气汤可能通过激活sirt1表达抑制FOXO1转录,进而降低炎症反应,减轻LPS诱导的大鼠急性肺损伤。

Objective:To investigate the effect of Xuanbai Chengqi Decoction for lipopolysaccharide(LPS)-induced acute lung injury in rats and its possible mechanism.Methods:50 clean SD male rats were injected with LPS into tail vein to establish acute lung injury model and randomly divided into model group,positive controlled group(Dexamethasone),low dose Xuanbai Chengqi Decoction group(2g crude drug/kg),medium dose Xuanbai Chengqi Decoction group(5g crude drug/kg)and high dose Xuanbai Chengqi Decoction group(8g crude drug/kg),with 10 rats in each group,and another 10 rats were set up as the normal controlled group.After the model was successfully prepared,the drug was administered continuously for 7 days,the pathological changes of lung tissue were observed by HE staining after the last administration,the protein content of alveolar lavage fluid was measured by Coomassie brilliant blue method,the levels of serum TNF-α and NO were measured by ELISA,and Western blotting(WB)was used to detect sirt1/FOXO1 pathway related protein expression in rat lung.Results:In the model group,the alveoli were filled with inflammatory exudates and bleeding,while in the low,medium and high Xuanbai Chengqi Decoction groups,the infiltration of inflammatory cells and bleeding were improved,compared with the normal group,the alveolar lavage fluid protein,lung volume index,levels of TNF-α,NO in serum,and FOXO1 protein in the lung tissue of the model group were significantly higher(P<0.05),and the level of sirt1 protein in the lung tissue of the model group was significantly lower(P<0.05).Compared with the model group,the alveolar lavage fluid protein,lung volume index,levels of TNF-α,NO in serum,and FOXO1 protein in the lung tissue of the low,middle and high dose Xuanbai Chengqi Decoction groups were all decreased in turn(P<0.05),the level of sirt1 protein in lung tissue of rats increased in turn(P<0.05),while there was no significant difference between the high dose Xuanbai Chengqi Decoction group and the positive controlled group in the alveolar lavage fluid protein,lung volume index,levels of TNF-α,NO in serum,and FOXO1 protein,sirt1 protein in the lung tissue(P>0.05).Conclusion:Xuanbai Chengqi Decoction may inhibit FOXO1 transcription by activating sirt1 expression,and then reduce the inflammatory response,and reduce LPS induced acute lung injury in rats.