Objective

2

To compare the effectiveness and mechanism of

Xiangzhu Fanggan Formula

（香术防感方） by sniffing and nasal drops for preventing influenza A H1N1flu.

Methods

2

Fifty-six BALB/c mice were randomly divided into normal group， model group， zanamivir group， high-concentration sachet group， low-concentration sachet group， high-concentration nasal drops group， and low-concentration nasal drops group， with 8 mice in each group. In the low- and high-concentration sachet gr

oups， 15 g and 30 g of

Xiangzhu Fanggan Formula

sachet were used for sniffing for 24 h per day； while in the low- and high-concentration nasal drops groups， nasal drops of

Xiangzhu Fanggan Formula

were given at a concentration of 0.11 and 0.22 g/ml， 20 μl each time， twice a day； in the zanamivir group， zanamivir was given at a concentration of 1.025 mg/ml of 20 μl each time， twice a day； in the normal group and the model group， nasal drops of normal saline were given at 20 μl each time， twice a day. Each group was given prophylactic intervention for 5 days. On day 5， 1 h after the administration of the drug， the mice in all groups except the normal group received 35 μl of 50 LD

50

A/PR/8/34/H1N1 viral solution as nasal drops to prepare influenza A H1N1 model mice. The body mass of the mice was recorded and the rate of change of body mass was calculated daily from day 5 to day 9 of the experiment， and the general status was observed. The mice were sampled on day 9， and the lung index and the inhibition rate of lung index were calculated； HE staining was used to detect pathological changes in lung tissues and to score lung tissue lesions； RT-qPCR was used to detect viral load in lung tissues； and ELISA was used to detect secretory immunoglobulin A （sIgA） and serum tumour necrosis factor α （TNF-α） and interleukin 2 （IL-2）， interleukin 6 （IL-6）， and interferon γ （IFN-γ） in the lavage fluid of the upper respiratory tract.

Results

2

On days 7， 8 and 9 of the experiment， the rate of change in body mass of mice in the model group significantly lower than that in the normal group at the same time points （

P

＜0.05 or

P

＜0.01）. On days 8 and 9 of the experiment， the rate of change in body mass of mice in the zanamivir group and the high-concentration nasal drops group increased when compared with the model group （

P

＜0.05 or

P

＜0.01）. Compared with the normal group， mice in the model group h

ad significantly higher lung index， lung tissue lesion score， lung tissue viral load， significantly higher serum TNF-α， IL-6， IL-2， IFN-γ levels， and significantly lower sIgA levels in the upper respiratory lavage fluid （

P

＜0.01）. Compared with the model group， the lung index and lung tissue viral load reduced， serum IFN-γ， TNF-α， IL-2， IL-6 levels reduced， and sIgA levels increased in the zanamivir group and the high-concentration nosal drops group （

P

＜0.05 or

P

＜0.01）； except for low-concentration sachet group， lung tissue lesion scores of the drug intervention groups reduced compared with those of the model group （

P

＜0.01）. Compared with the zanamivir group， the lung index increased in the low-concentration sachet group and the low- and high-concentration nasal drops groups， and the serum TNF-α and IL-2 levels increased in all

Xiangzhu Fanggan Formula

intervention groups （

P

＜0.05 or

P

＜0.01）. Compared with high-concentration nasal drops group， serum TNF-α and IFN-γ levels elevated in the high-concentration increased group， and lung tissue viral load elevated in the low-concentration nasal drops group （

P

＜0.05 or

P

＜0.01）. The lung index inhibition rate was 80.84% in the zanamivir group， 41.61% and 17.90% in the high- and low-concentration sachet groups， and 35.40% and 25.40% in the high- and low-concentration nasal drops groups， respectively. HE staining showed that the lung tissues of the model group showed thickening of alveolar septa， alveolar collapse， and infiltration of inflammatory cells； whereas， in each drug intervention group， the inflammation of the lung tissues of the mice and the damage reduced， and the most obvious improvement was in the zanamivir group and the high-concentration nasal drops group.

Conclusion

2

Xiangzhu Fanggan Formula

by sniffing and nasal drops could both prevent influenza A H1N1 virus infe

ction， with antiviral and anti-inflammatory effects， also could improve the pathological damage of lung tissue， and improve the immunity of respiratory mucosa. The nasal drops may be better than sachets in inhibiting inflammatory response， especially the high-concentration nasal drops showed more effective.