鬼针草调节PI3K-Akt信号通路改善急性肺损伤作用的分子机制研究

Study on the effect and molecular mechanism of Bidens bipinnata L.on improvement of acute lung injury by regulating PI3K-Akt signal pathway

目的研究鬼针草对脂多糖(LPS)诱导急性肺损伤(ALI)的改善作用及其机制。方法将60只Balb/c小鼠随机分为6组:空白组、模型组、鬼针草(低、中、高剂量)组及阳性药组,腹腔注射LPS诱导ALI模型12 h后进行灌胃给药,建模5 d后处死并分离肺组织,观察组织病理切片及检测相关炎症因子活性,通过网络药理学预测鬼针草改善ALI作用的药效物质基础及潜在的作用机制,采用免疫组织化学方法对相应通路的关键靶点进行验证。结果鬼针草可改善ALI小鼠肺泡间质水肿、肺泡壁增厚等病理损伤,显著抑制肺组织髓过氧化物酶(MPO)及血清TNF-α,IL-1β和IL-6的表达(P<0.05),从而缓解机体的炎症反应。通过网络药理学预测分析,鬼针草治疗ALI的主要活性成分可能为黄酮类化合物中的槲皮素和木犀草素;通过作用于TNF、IL-6、ALB、GAPDH、AKT1、VEGFA、STAT3等靶点,调控PI3K-Akt、TNF、HIF-1等通路来发挥改善ALI的作用。RT-qPCR及免疫组织化学结果显示鬼针草可促进ALI小鼠肺组织Akt的转录及表达(P<0.05)。结论鬼针草对ALI炎症反应具有改善作用,其机制可能为激活PI3K-Akt信号通路来发挥肺保护作用。为鬼针草的进一步开发利用提供了一定的实验基础。

Objective To investigate the improving effect and mechanism of Bidens bipinnata L.on LPSinduced acute lung injury(ALI).Methods Sixty Balb/c mice were randomly divided into six groups,including the blank group,the model group,and low,medium and high doses of Bidens bipinnata L.groups and positive drug group.The ALI model was induced by intraperitoneal injection of LPS for 12 hours,and the lung tissue was collected 5 days after modeling.The histopathological changes were observed and the activities of inflammatory factors were detected.The pharmacological material basis and potential mechanism of Bidens bipinnata L.improving ALI were predicted by network pharmacology.The key targets of the corresponding pathway were measured by immunohistochemical method.Results Bidens bipinnata L.ameliorated the pathological damage such as alveolar interstitial edema and alveolar wall thickening in ALI mice,and inhibited the expression of MPO in lung tissue and the levels of TNF-α,IL-1βand IL-6 in serum(P<0.05),thereby alleviating the body's inflammatory response.Through the prediction and analysis of network pharmacology,the main active components of Bidens bipinnata L.in treating ALI might be quercetin and luteolin in flavonoids,and improved ALI by acting on targets such as TNF,IL-6,ALB,GAPDH,AKT1,VEGFA,STAT3 and regulating PI3K-Akt,TNF,HIF-1 pathways.RT-qPCR and immunohistochemistry results showed that Bidens bipinnata L.promoted the transcription and expression of Akt in lung tissue of ALI mice(P<0.05).Conclusion Bidens bipinnata L.can improve the inflammatory reaction of ALI,and its mechanism may be to activate PI3K-Akt signal to protect the lung tissue,which provides experimental support for the further development and utilization of Bidens bipinnata L.