Objective

2

To explore the mechanism of Jingyin Shuangjie Powder （JYSJ） in treating influenza A virus infection in mice based on the phosphoinositide 3-kinase （PI3K）/protein kinase B （AKT） signaling pathway.

Methods

2

A total of 96 mice were randomly divided into the blank group， model group， western medicine group， and Chinese medicine group according to a random number table， with 24 mice in each group for pharmacodynamics study. Another 25 mice were randomly divided into the blank group， model group， Chinese medicine group， pathway agonist group， and pathway agonist combined with Chinese medicine group according to a random number table， with 5 mice in each group for the mechanism study. Except for the blank group， all other groups were established with a model of influenza virus （Asian type mouse lung-adapted strain PR8） infection. Mice in the Chinese medicine group were administered JYSJ ［3 mg/（g·d）］ via intragastric administration， mice in the western medicine group were given oseltamivir phosphate ［22.75 mg/（kg·d）］ via intragastric administration， mice in the pathway agonist group were given insulin-like growth factor-1 （IGF-1） ［50 μg/（kg·d）］ via intraperitoneal injection， and mice in the pathway agonist combined with Chinese medicine group were administered IGF-1 ［50 μg/（kg·d）］ via intraperitoneal injection plus JYSJ ［3 mg/（g·d）］ via intragastric administration， once per day. In the pharmacodynamics study， after 1， 3， and 5 days of continuous administration， the general condition， lung index， lung index inhibition rate， and pathological changes in lung tissue were compared. The levels of IL-6， IL-10， and IFN-γ in lung tissue were detected by ELISA. In the mechanism study， after 5 days of continuous administration， the relative expression levels of PI3K， p-AKT， AKT， and NF-κB p65 proteins in lung tissue were detected by Western blot， and the mRNA expression levels of PI3K， AKT， and NF-κB p65 were detected by RT-qPCR.

Results

2

Compared with the model group， the general condition of the mice in the Chinese medicine group was better， with higher body weight and food intake. The lung index was lower on days 3 and 5 （

P

<

0.05）， and lung tissue inflammatory infiltration and erythrocyte exudation were lighter. Compared with

the model group， the relative expression level of IL-6 in lung tissue of the Chinese medicine group was lower on day 3 （

P

<

0.05）， while the relative expression level of IL-10 was higher on day 3 （

P

<

0.05）. The relative expression of IFN-γ in lung tissue was lower on days 3 and 5 after administration （

P

<

0.05）. Compared with the model group， the relative expression levels of PI3K， p-AKT/AKT， and NF-κB p65 proteins in the Chinese medicine group were lower （

P

<

0.05）， and the relative expression levels of PI3K， AKT， and NF-κB mRNA were also lower （

P

<

0.05）. Compared with the Chinese medicine group， the relative expression levels of PI3K， p-AKT/AKT， NF-κB p65 proteins， and the mRNA expression of PI3K， AKT， and NF-κB p65 were significantly higher in the pathway agonist combined with Chinese medicine group （

P

<

0.05）.

Conclusion

2

JYSJ can improve the general condition of mice infected with influenza A virus， reduce the lung index， improve cytokine expression， and alleviate lung pathological damage. Its mechanism may involve the downregulation of key proteins and gene expression in the PI3K/AKT signaling pathway.