葶苈子水提液通过调控HMGB1/TLR4/NF-κB信号通路对LPS吸入性肺损伤新生大鼠的影响

Effect of Semen Lepidii aqueous extract on neonatal rats with LPS inhalation lung injury by regulating HMGB1/TLR4/NF-κB signaling pathway

[目的]探究葶苈子水提液对脂多糖(LPS)吸入性肺炎新生大鼠的影响。[方法]腹腔注射LPS制备吸入性肺损伤大鼠模型,将大鼠随机分为6组:对照组、模型组、葶苈子水提液5 g/kg组、葶苈子水提液10 g/kg组、葶苈子水提液20 g/kg组和地塞米松0.002 g/kg组。干湿法检测肺组织干湿质量比值;苏木精-伊红(HE)染色检测肺组织病理损伤;酶联免疫吸附(ELISA)法检测氧化应激标记物:超氧化物歧化酶(SOD)、丙二醛(MDA)、谷胱甘肽(GSH)含量和炎症因子白细胞介素-6(IL-6)、白细胞介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)水平;蛋白免疫印迹(Western Blot)法检测高迁移率族蛋白1(HMGB1)、Toll样受体4(TLR4)和磷酸化核转录因子-κB(p-NF-κB)p65蛋白表达水平。[结果]与对照组比较,模型组大鼠肺干湿质量比值升高,SOD、GSH含量降低,MDA含量升高,IL-6、IL-1β和TNF-α水平升高,HMGB1、TLR4蛋白表达水平及p-NF-κB p65与核转录因子-κB(NF-κB)p65比值升高,差异有统计学意义(P<0.05)。与模型组比较,葶苈子水提液10、20 g/kg组和地塞米松0.002 g/kg组肺干湿质量比值降低,肺组织病理损伤程度明显改善,SOD、GSH含量升高,MDA含量降低,IL-6、IL-1β和TNF-α水平降低,HMGB1、TLR4蛋白表达水平及p-NF-κB p65与NF-κB p65比值降低,差异有统计学意义(P<0.05)。[结论]葶苈子水提液能改善LPS诱导的吸入性肺损伤大鼠模型病理损伤,抑制炎症因子的释放,改善氧化应激,抑制HMGB1/TLR4/NF-κB信号通路的活化。

[Objective]To investigate the effect of Semen Lepidii aqueous extract on neonatal rats with LPS aspiration pneumonia.[Methods]Rat models of aspiration lung injury were prepared by intraperitoneal injection of liposaccharide(LPS).The rats were randomly divided into 6 groups:control group,model group,water extract of Semen Lepidii(5 g/kg),water extract of Semen Lepidii(10 g/kg),water extract of Semen Lepidii(20 g/kg),and dexamethasone(0.002 g/kg).The ratio of dry and wet lung tissue was measured by dry and wet method.HE staining was used to detect pathological damage of lung tissue.The levels of superoxide dismutase(SOD),malondialdehyde(MDA)and glutathione(GSH)and inflammatory cytokines interleukin-6(IL-6),interleukin-1β(IL-1β)and tumor necrosis factor-α(TNF-α)were detected by ELISA.The high mobility group protein B1(HMGB1),Toll like receptor 4(TLR4)and p-NF-κB p65 protein expression levels were detected by Western Blot.[Results]Compared with the control group,there were statistically significant differences between the model group and the control group(P<0.05)in increased lung dry-wet weight ratio,increased levels of SOD,MDA and GSH,increased levels of IL-6,IL-1βand TNF-α,and increased expression levels of HMGB1 and TLR4 proteins,and increased p-NF-κB p65/NF-κB p65.Compared with model group,by the Semen Lepidii water extraction liquid of 10 g/kg,20 g/kg,0.002 g/kg group and dexamethasone group reduced lung wet weight ratio,lung tissue pathology damage degree,SOD and GSH content increased,MDA content reduced,IL-6,IL-1βand TNF-αlevels reduced,HMGB1,TLR4 protein expression level and p-NF-κB p65/NF-κB p65 ratio decreased,significant difference was statistically significant(P<0.05).[Conclusion]Semen Lepidii aqueous extract can improve the pathological injury,inhibit the release of inflammatory factors,improve oxidative stress,and inhibit the activation of HMGB1/TLR4/NF-κB signaling pathway in the rat model of inhalation lung injury induced by LPS.