金振口服液对LPS致急性肺损伤模型小鼠NF-κB,MAPK信号通路的影响

Effect of Jinzhen Oral Liquid on NF-κB,MAPK Signaling Pathway in Mice with LPS-induced Acute Lung Injury

目的：探讨金振口服液（JZKFY）对脂多糖（LPS）致急性肺损伤（ALI）模型小鼠的影响及其分子水平作用机制。方法：小鼠随机分为正常组、模型组、醋酸地塞米松组、金振口服液高、中、低（4.4,2.2,1.1 g·kg（-1））剂量组,各给药组给予相应剂量药物,正常组和模型组灌胃同等剂量的生理盐水,1次/d,连续7 d后,模型组及药物组小鼠腹腔注射LPS（10 mg·kg（-1））复制急性肺损伤小鼠模型,正常组腹腔注射同等剂量的生理盐水。6 h后采集小鼠肺组织,测定左肺湿/干质量比（W/D）;酶联免疫吸附法（ELISA）检测肺组织中肿瘤坏死因子-α（TNF-α）,白细胞介素（IL）-1β;蛋白免疫印迹法（Western blot）测定肺组织中p65,IκBα,ERK1/2,p38蛋白及其磷酸化蛋白表达水平。结果：与正常组比较,模型组小鼠W/D显著升高（P〈0.01）,肺组织中TNF-α,IL-1β水平显著升高（P〈0.01）,p65,IκBα,ERK1/2,p38蛋白磷酸化水平显著上调（P〈0.01）。与模型组比较,金振口服液低、高剂量及地塞米松组W/D显著降低（P〈0.05,P〈0.01）;金振口服液各剂量组及地塞米松组肺组织中TNF-α,IL-1β水平显著降低;金振口服液各剂量组及地塞米松组p65,IκBα,p38蛋白磷酸化水平显著下调（P〈0.05,P〈0.01）,金振口服液高剂量、地塞米松组ERK1/2蛋白磷酸化水平显著下调（P〈0.05,P〈0.01）。结论：金振口服液能够有效改善LPS诱导的ALI肺组织间质性水肿及降低致炎细胞因子的含量,其作用机制与抑制p65,IκBα,ERK1/2,p38多个靶蛋白磷酸化,从而阻断核转录因子-κB（NF-κB）,丝裂原活化蛋白激酶（MAPK）炎症通路的信号传导密切相关。

Objective：To explore the effect and mechanism of Jinzhen oral liquid（JZOL） on lipopolysaccharide（LPS）-induced acute lung injury（ALI） model in mice.Method：Mice were randomly divided into normal control group,model group,dexamethasone group（DEX）,JZOL high（JZOLG）,medium（JZOLZ）and low（JZOLD） dose（4.4,2.2,1.1 g·kg^（-1）） groups.Corresponding dose of medicine was given in the treatment groups,and the mice in normal group and model group received the same dose of normal saline,once a day for 7 days.Then the ALI models were induced by intraperitoneal injection of LPS（10 mg·kg^（-1））,while the mice in normal group received intraperitoneal injection of the same dose of normal saline.Six hour later,the lung tissues were taken; left lung wet-to-dry ratio（W/D） was measured; tumor necrosis factor（TNF）-α,and interleukin（IL）-1β in lung tissues were detected by enzyme-linked immunosorbent assay（ELISA）; the protein expression levels of p65,IκBα,ERK1/2,p38 protein and their phosphorylation levels in lung tissues were detected by Western blot.Result：As compared with the normal group,the W/D,levels of TNF-α,IL-1β and the phosphorylation expression levels of p65,IκB,p38 protein were increased in the model group（P 〈 0.01）.As compared with the model group,the W/D was significantly decreased in JZOLD,JZOLG and DEX groups; the levels of TNF-α and IL-1β were decreased significantly,and the phosphorylation expression levels of p65,IκB,p38 protein were significantly down-regulated in JZOLG,JZOLZ,JZOLD,and DEX groups（P 〈 0.05,P 0.01）; and the phosphorylation expression level of ERK1/2 was significantly down-regulated in JZOLG and DEX groups（P 〈 0.05,P 〈 0.01）.Conclusion：JZOLY can effectively improve the interstitial edema of the lung tissues and reduce inflammatory cytokines in mice with LPS-induced ALI.The mechanism is closely associated with inhibiting the phosphorylation levels of p65,IκBα,ERK1/2 and p38,blocking nuclear transcription factor-kappa B（NF-κB） and mitogen activated protein kinase（MAPK） inflammatory pathway.