抗奥合剂通过p38 MAPK/NF-κB信号通路和ACE2/Ang1-7/Mas轴缓解急性肺损伤研究

Kangaoheji Relieves Acute Lung Injury through p38 MAPK/NF-κB Signaling Pathway and ACE2/Ang1-7/Mas Axis

目的探讨抗奥合剂(KAHJ)治疗小鼠急性肺损伤(ALI)的作用及机制,为其可能作为缓解新型冠状病毒(COVID-19)感染后症状的药物提供依据。方法采用网络药理学方法预测KAHJ治疗ALI的主要活性成分、潜在靶点和相关信号通路。将C57BL/6J小鼠随机分为对照组、LPS组和LPS+KAHJ组。LPS+KAHJ组小鼠灌胃KAHJ(4.76 g·kg^(-1)·d^(-1),8.8 mL·kg^(-1)·d^(-1)),其余组小鼠灌胃生理盐水(8.8 mL·kg^(-1)·d^(-1))。14 d后,腹腔注射LPS(5 mg·kg^(-1))诱导ALI模型。收集小鼠血清和肺组织,通过组织病理学观察肺组织的病理变化。采用Western blot、qPCR、ELISA和IHC等方法评估KAHJ对ALI的改善作用。结果通过网络药理学筛选出疾病和药物共同的70个核心靶基因,并显示与多个信号通路密切相关,如MAPK、NF-κB、Apoptosis、COVID-19和肾素-血管紧张素系统(Ras)信号通路等。此外,通过实验验证发现KAHJ能改善小鼠ALI后的炎症和细胞凋亡,减少肺损伤和肺水肿,抑制肺纤维化。同时,KAHJ的作用机制与p38 MAPK和NF-κB的磷酸化以及ACE2/Ang1-7/Mas轴的调控也有着密切关系。结论KAHJ可能通过抑制p38 MAPK/NF-κB信号通路和调控ACE2/Ang1-7/Mas轴缓解ALI,为缓解COVID-19感染后症状提供了补充和替代药物。

OBJECTIVE To explore the effect and mechanism of Kangaoheji(KAHJ)in the treatment of acute lung injury(ALI)in mice,and provide a rationale for its possible use as a drug to alleviate symptoms following coronavirus disease 2019(COVID-19)infection.METHODS Network pharmacology was carried out to predict the main active components and potential targets of KAHJ on ALI.C57BL/6J mice were randomly divided into Control group,LPS group and LPS+KAHJ group.LPS+KAHJ group was gavaged with KAHJ(4.76 g·kg^(-1)·d^(-1),8.8 mL·kg^(-1)·d^(-1))and the rest of the groups were gavaged with saline(8.8 mL·kg^(-1)·d^(-1)).LPS(5 mg·kg^(-1))was injected intraperitoneally to induce an acute inflammation model after 14 d.The serum and lung tissues of mice were collected,and the pathological changes in lung tissues were observed via histopathology.Western blot,Real-time PCR,Enzyme-Linked immunosorbent assay(ELISA)and immunohistochemistry(IHC)were used to assess the ameliorative effect of KAHJ on ALI.RESULTS The result showed that 70 core target genes of KAHJ on ALI were primarily implicated in multiple signaling pathways involving MAPK signaling pathway,NF-κB signaling pathway,apoptosis,and Ras signaling pathway.Furthermore,we found that KAHJ ameliorated inflammation and apoptosis in ALI,thereby reducing lung damage and pulmonary edema and inhibiting pulmonary fibrosis.Additionally,KAHJ inhibited the phosphorylation of p38 MAPK and NF-κB,and upregulated the ACE2/Ang1-7/Mas axis.CONCLUSION KAHJ might relieve acute lung injury through p38 MAPK/NF-κB signaling pathway and ACE2/Ang1-7/Mas axis,which offers complementary and alternative treatment options for COVID-19.