参附注射液抑制HMGB1核转位保护内毒素休克大鼠肺损伤

Effect of Shenfu injection on endotoxin shock induced lung injury rats by inhibiting the nuclear translocation of HMGB1

目的:探讨参附注射液保护内毒素休克大鼠肺损伤的分子机制。方法:经腹腔注射脂多糖(LPS)建立内毒素休克SD大鼠模型,用5ml/kg、10ml/kg、15ml/kg剂量参附注射液干预,观察肺组织病理改变,检测肺组织中高迁移率族蛋白B1(HMGB1)的转录、表达、亚细胞定位及血浆中HMGB1分泌水平。结果:LPS可致SD大鼠肺泡隔水肿,充血,大量粒细胞浸润;肺组织中HMGB1的转录和表达均显著上调;胞浆中HMGB1表达显著增加,而核内HMGB1则显著下调;血浆中HMGB1水平明显上升。与模型组比较,参附注射液各剂量均可减轻内毒素休克大鼠肺组织水肿、充血和炎性细胞渗出,对肺损伤具有明显的保护作用,10ml/kg组病理评分最低;抑制内毒素休克大鼠肺组织中HMGB1的转录(呈剂量依赖性)、翻译和核转位(以10ml/kg组效果最优);抑制HMGB1分泌出胞,减少血浆中HMGB1含量(呈剂量依赖性)。结论:参附注射液抑制内毒素休克大鼠肺组织中HMGB1的核转位,可能是其扶阳固脱的重要分子机制。

Objective: To investigate the molecular mechanism of Shenfu Injection on protecting lung injury in endotoxin shock rats. Methods: Lipopolysaccharide( LPS) was intraperitoneally injected to SD rats to establish the endotoxin shock model. Animals were administered with Shenfu injection( 5 ml/kg、10 ml/kg、15 ml/kg doses). The pathological changes of lung tissues were observed. The transcription,expression,subcellular localization of high mobility group protein( HMGB1) in lung tissues and the secretion of HMGB1 in plasma were detected. Results: Thickened alveolar septa,pulmonary capillary hyperemia and notable inflammatory cell infiltration were readily identified after a stimulation by LPS. The transcription and expression of HMGB1 in lung tissues were significantly up regulated in LPS group. LPS markedly increased cytoplasmic HMGB1,and decreased nuclear HMGB1. The secretion of HMGB1 in plasma obviously increased in LPS group. Compared with LPS group,SFI reduced pathological changes in lung tissues( The pathological score was the lowest in 10 ml/kg group),downregulated the transcription( in dose-dependent manner),the expression and the translocation of HMGB1 in the cells of lung tissues( 10 ml/kg dose group had the best effect),as wells as the secretion of HMGB1 in plasma( in dose-dependent manner). Conclusion: Shenfu injection can significantly inhibit the nuclear translocation of HMGB1 in lung tissues and the secretion of HMGB1 in plasma of endotoxin shock rats,which may be an important molecular mechanism of its Fuyanggutuo effect.