OBJECTIVE

2

To identify the quality markers （Q-Markers） of

Hyssopus cuspidatus

against airway remodeling in bronchial asthma （referred to as “asthma”）， an

d to provide a reference for the quality control research of

H. cuspidatus

based on pharmacodynamic activity.

METHODS

2

Potential active components and action targets of

H. cuspidatus

were screened by network pharmacology method. Using human airway smooth muscle cells （HASMCs） as objects， airway remodeling cell model was induced by platelet-derived growth factor-BB （PDGF-BB）； validation test was then performed for anti-asthmatic effects of

H. cuspidatus

and the potential active components. HPLC method was employed to establish the fingerprints of 14 batches of

H. cuspidatus

samples， and chemometric analysis was also conducted. Combined with the results of pharmacodynamic experiments and fingerprint analysis， the Q-Markers of

H. cuspidatus

against airway remodeling in asthma were determined.

RESULTS&CONCLUSIONS

2

Network pharmacology analysis showed that the potential active components of

H. cuspidatus

against asthma might be flavonoids and phenolic acids such as luteolin， quercetin and rosmarinic acid， and the core anti-asthmatic targets were interleukin-6， mitogen-activated protein kinase， etc.

In vitro

experimental results confirmed that 25， 50， 100 μg/mL of

H. cuspidatus

， as well as neochlorogenic acid （80 μmol/L）， acacetin （80 μmol/L）， salvianolic acid B （40 μmol/L） and quercetin-3-

O

-

β

-D-glucuronide （80 μmol/L）， significantly reduced the cell viability induced by PDGF-BB， inhibited cell proliferation， migration， and the phosphorylation level of extracellular signal-regulated protein kinase 1/2， decreased the levels of interleukin-6， tumor necrosis factor-α and reactive oxygen species， and generally arrested cells in the G

0

/G

1

phase （

P

＜0.05）. Fingerprint analysis showed that there were 27 common peaks in the fingerprints of the 14 batches of

H. cuspidatus

samples， with 15 compounds （including luteolin） identified， and the similarities of fingerprints were all greater than 0.8. The 14 batches of samples could be divided into three categories： S1-S7 as one category， S8-S13 as one category， and S14 as one category. The variable importance in the projection values of rosmarinic acid， chlorogenic acid， caffeic acid， salvigenin， luteolin， ferulic acid， quercetin-3-

O

-

β

-D-glucuronide， and the components corresponding to peaks 5 and 8 were greater than 1， indicating they were potential differential markers affecting quality. Integrating network pharmacology，

in vitro

experimental validation， and chemometric analysis， rosmarinic acid， neochlorogenic acid， caffeic acid， salvigenin， luteolin， ferulic acid， quercetin-3-

O

-

β

-D-glucuronide， acacetin， salvianolic acid B and chlorogenic acid may be the Q-Markers of

H. cuspidatus

against asthma.