Study on the active components and mechanism of Callicarpa nudiflora against H1N1 based on UPLC-Q-Exactive-Orbitrap-MS,network pharmacology and experimental validation
In this experiment,the extracts of Callicarpa nudiflora were extracted to obtain different extraction parts,and then the sites were screened for anti-influenza A virus H1N1(H1N1)activity,and it was determined that the ethyl acetate extrac-tion part of Callicarpa nudiflora(EACN)had better anti-H1N1 activity in vitro.Qualitative analysis of EACN by ultra per-formance liquid chromatography-quaternary/orbitrap-high resolution mass spectrometry(UPLC-Q-Exactive-Orbitrap-MS)technique was performed.Based on the results of qualitative analysis,data mining and network pharmacology were further used to evaluate the active ingredients,key targets and critical pathways of the drug.Finally,the prediction results were validated u-sing the H1N1 infected BALB/c mouse model.The results showed that the ethyl acetate extract had the best anti-H1N1 effect in vitro,with half inhibitory concentration(IC50)of 51 μg/mL,half cytotoxic concentration(CC50)of 859.4 μg/mL,and se-lection index(SI,CC50/IC50)value of 16.85.Thirty-four compounds were identified from EACN.On this basis,67 drug tar-gets related to EACN anti-H1N1 were screened using network pharmacology,including 491 biological processes and 147 path-ways related to anti-H1N1.The experimental results of H1N1 infected BALB/c mice showed that EACN could improve the weight loss and lung index increase caused by H1N1 infection,reduce the viral load of mouse lung tissue,alleviate the patho-logical injury of mouse lung tissue,and reduce the levels of TNF-α,IFN-γ and IL-1β in peripheral blood serum.The results suggest that EACN may play an anti-H1N1 role in many ways,such as direct inhibition of virus,anti-inflammatory effect and immunomodulatory effect.