丹参酮ⅡA激活Nrf2/Nox4通路减轻脂多糖诱导的小鼠肺炎和纤维化

Tanshinone ⅡA attenuates LPS-induced pneumonia and fibrosis in mice via activating Nrf2/Nox4 pathway

目的研究丹参酮ⅡA(Tan ⅡA)对脂多糖诱导小鼠肺炎和纤维化影响,探讨其潜在的作用机制。方法30只8周龄雄性C57BL/6小鼠,随机分成对照组、脂多糖(LPS)组和LPS+Tan ⅡA组。采用尾静脉注射LPS制作小鼠肺炎和纤维化动物模型,术后给予Tan ⅡA干预,每日一次,连续2周,实验第4周处死所有小鼠,收集血清和肺组织标本。采用HE染色检测肺组织炎症改变,Masson染色观察肺组织纤维化程度,ELISA检测血清IL-1β、TNF-α和IL-10蛋白表达,采用Western blot技术检测肺组织Nrf2、Nox4、Nqo1和Ho-1蛋白表达。结果与对照组相比,LPS组小鼠肺脏重量、肺脏/体重比显著增加,肺组织水肿、炎细胞浸润和纤维化明显加重,血清促炎蛋白IL-1β、TNF-α显著升高,抑炎蛋白IL-10表达下降(P<0.05)。Tan ⅡA处理显著减轻LPS导致的小鼠肺脏重量、肺脏/体重比,减轻肺水肿、炎细胞浸润和纤维化,减低血清IL-1β、TNF-α表达水平,升高IL-10表达水平。此外,Nrf2和Nox4蛋白表达水平LPS组高于对照组,Nqo1和Ho-1蛋白表达水平低于对照组,而Tan ⅡA处理可以上调Nrf2、Nqo1和Ho-1蛋白,降低Nox4蛋白表达水平(P<0.05)。结论Tan ⅡA减轻LPS诱发的小鼠炎症和纤维化,与激活Nrf2/Nox4通路相关。

Objective To investigate the effect of Tanshinone ⅡA(Tan ⅡA)on lipopolysaccharide-induced inflammation and fibrosis in mice,and its potential mechanism.Methods Thirty male C57 BL/6 mice aged 8 weeks were randomly divided into control group,lipopolysaccharide(LPS)group and LPS+Tan ⅡA group.The model of inflammation and fibrosis in mice was established by tail vein injection of LPS.Tan ⅡA was given to the mice once a day for 2 weeks.All mice were killed at the fourth week of the experiment,serum and lung tissue samples were collected.HE staining was used to detect inflammatory changes in lung tissue,Masson staining was used to observe the degree of pulmonary fibrosis,ELISA was used to detect the expression of IL-1 beta,TNF-alpha and IL-10 in serum,and Western blot was used to detect the expression of Nrf2,Nox4,Nqo 1 and Ho-1 in lung tissues.Results Compared with the control group,the lung weight,lung/body weight ratio,pulmonary edema,inflammatory cell infiltration and fibrosis were significantly increased in LPS group,and the levels of serum pro-inflammatory protein IL-1 beta and TNF-alpha were significantly increased,while the expression level of anti-inflammatory protein IL-10 was significantly decreased.Tan ⅡA treatment significantly reduced LPS-induced lung weight and lung/body weight ratio and pulmonary edema,inflammatory cell infiltration and fibrosis,decreased the expression level of serum IL-1 beta and TNF-alpha,and increased the expression level of IL-10.The expression level of Nrf2 and Nox4 proteins in lung tissue increased,but decreased for Nqo1 and Ho-1 expressions in LPS group than in control.However,Tan ⅡA treatment significantly increased the expression of Nrf2 and Nqo1,Ho-1 protein,and decreased the expression of Nox4 protein(P<0.05).Conclusion Tan ⅡA treatment alleviated LPS-induced inflammation and fibrosis,which is related to the activation of Nrf2/Nox4 pathway in mice.