麻杏解毒合剂通过p38MAPK/AP-1通路对冠状病毒肺炎模型小鼠炎性因子的调控作用研究

Study on the Regulatory Effect of Maxing Jiedu Mixture on Inflammatory Factors in Coronavirus Pneumonia Model Mice Based on the p38MAPK/AP-1 Pathway

目的研究麻杏解毒合剂对冠状病毒肺炎模型小鼠炎性因子表达的影响,基于p38MAPK/AP-1通路研究其疗效机制。方法60只昆明种(KM)小鼠随机分成空白对照组、模型组、p38MAPK抑制剂组及麻杏解毒合剂高、中、低剂量组,每组10只。采用模拟寒湿环境、表达h ACE2的重组腺相关病毒转导、SARS-CoV-2spike假病毒气管内给药建立小鼠冠状病毒肺炎模型。检测各组小鼠血清炎性因子、肺组织病理改变、肺组织p38MAPK、c-jun、c-fos的mRNA水平和蛋白表达情况。结果与空白对照组比较,模型组小鼠血清炎性因子白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)、白细胞介素-8(IL-8)、肿瘤坏死因子-α(TNF-α)的表达均显著升高(P<0.01),肺组织炎性改变明显,c-fos m RNA水平和p-p38、c-fos、c-jun蛋白表达均显著增加(P<0.05)。与模型组比较,麻杏解毒合剂高、中剂量组小鼠血清炎性因子显著降低(P<0.01或P<0.05),小鼠肺组织炎性损伤明显减轻,同时肺组织中c-fos的mRNA水平和p-p38、c-fos的蛋白表达均显著下调(P<0.05)。结论麻杏解毒合剂能降低病毒性肺炎模型小鼠血清炎性因子水平,减轻肺组织炎性损伤,其疗效机制与抑制p38MAPK蛋白的磷酸化,下调AP-1通路mRNA及蛋白表达有关。

Objective:To study the effect of Maxing Jiedu Mixture on the expression of inflammatory factors in coronavirus pneumonia model mice,and to investigate its therapeutic mechanism based on the p38MAPK/AP-1pathway.Methods:60 Kunming mice(KM)were randomly divided into blank control group,model group,p38MAPK inhibitor group,high,medium,and low dose groups of Maxing Jiedu Mixture,with 10 mice in each group.Coronavirus pneumonia model was established using simulated cold and humid environments,recombinant adenoassociated virus transduction expressing h ACE2,and intratracheal administration of SARS-CoV-2 spike pseudovirus.The serum inflammatory factors,pathological changes in lung tissue,mRNA levels and protein expression of p38MAPK,c-jun,and c-fos in lung tissue of mice in each group were detected.Results:Compared with the blank control group,the expressions of serum inflammatory factor IL-1β,IL-6,IL-8,TNF-α in the model group were significantly increased(P<0.01),and the inflammatory changes in lung tissue were significant,the expression level of c-fos mRNA and p-p38,c-fos,and c-jun proteins were significantly increased(P<0.05).Compared with the model group,the high and medium dose groups of Maxing Jiedu Mixture significantly reduced the serum inflammatory factors(P<0.01,P<0.05),and significantly reduced inflammatory damage to the mouse lung tissue,the mRNA of c-fos and the protein expression level of p-p38 and c-fos in the lung tissue were significantly downregulated(P<0.05).Conclusion:Maxing Jiedu Mixture can reduce the levels of serum inflammatory factors and alleviate lung tissue inflammatory damage in viral pneumonia model mice.Its therapeutic mechanism is related to inhibiting the phosphorylation of p38MAPK protein and downregulating the expression of AP-1 pathway mRNA and protein.