## 4 Discussion
In our data, suppression of ERK is more effective in lower doses of JGTW treatment, which is occasionally observed result that the lower dose of herbal extract is more effective in regulating signaling pathway. This might be due to the combinational effect of various compounds of JGTW; hence, further studies regarding the active compound responsible for inhibiting MAPK pathway should be done. OVA-sensitized and challenged mice represented typical asthmatic features such as increased inflammatory cell recruitment and elevated IgE level. In the present study, JGTW treatment after OVA sensitization effectively reduced the number of inflammatory cells in BALF, suggesting the effectiveness of anti-inflammatory response by JGTW shown in vitro on relieving symptoms of asthma in animal model. Additionally, the levels of IL-5 and IL-13 in BALF, a hallmark of Th2 response in asthma, which were elevated in response to the OVA treatment, showed a tendency to decrease by JGTW treatment. Furthermore, plasma levels of IgE, both total and OVA-specific, in JGTW treated mice were significantly lower than those of OVA-induced mice. Collectively, these results suggest that JGTW suppresses the inflammatory and allergic responses associated with asthma. As previously mentioned, JGT is composed of two herbal ingredients, _P. lactiflora_ and _G. unlensis_, and their chemical constituents mainly include flavonoids (e.g., flavones, flavanones, flavonoid glycosides), terpenoids (e.g., monoterepenes, monoterepene glycosides, triterpenenes), and chalcones [22,23]. In this study, UPLC-DAD-MS/MS analysis was performed to identify and quantify the 24 phytochemicals in JGTW, comprising 12 flavonoids (catechin, vicenin-2, schaftoside, isoschaftsoide, neoliquiritin, liquiritin, luiqiritin, luiteolin, calycosin, naringeni, kaempfer, and formonentin), three chalcones (licochalcone B, echinatin, and isoliquiritinen), seven terpenoids (albiflorin, paenoniflorin, mudanpioside C, benzoylpaeoniflorin, glycyrrhizin, hederagenin, and glycyrrheinic acid), and one phenol (1,2,3,4,6-O-pentagalloyglucose), and paeoniflorin (63.971 mg/g) from _P. lactiflora_ and glycyrrhizin (11.853 mg/g) from _G. unalensis_ were abundantly found in JGTW. The individual herbal compounds that constitute JGT have been reported to be effective against airway inflammation-related diseases, such as asthma. Paeoniflorin, a major component of _P. lactiflora_, exerts anti-asthmatic effects by modulating the Th1/Th2 equilibrium [37] and inhibiting the growth and migration of airway smooth muscle cells [38]. Total glucosides of _P. lactiflora_ improve ovalbumin-induced allergic asthma in mice by suppressing mast cell degranulation [39]. Glycyrrhizin, a major component of _G. unalensis_, alleviates asthmatic symptoms in mice [40] and ameliorates the chronic histopathological changes in the lungs of mice suffering from asthma [41] and asthmatic progression through modulation of Th1/Th2 cytokines and enhancement of CD4\({}^{+}\)CD25\({}^{+}\)Foxp3\({}^{+}\) Tregs [42]. In addition, components of _G. unalensis_, isoliquitiigenin and licochalcone A, relax guinea pig tracheal smooth muscle in vitro and in vivo [43] and alleviate oxidative stress and hyper-responsiveness in asthmatic mice [44], respectively. These previous studies and our quantitative analysis data for the phytochemicals in JGTW suggest that the anti-inflammatory activity of JGTW in IT-stimulated BEAS-2B and LPS-stimulated RAW264.7 cells may be attributed to paeoniflorin and glycyrrhizin. In summary, our results discovered that JGTW might exert anti-inflammatory effects by suppressing the airway inflammation-related factors in pulmonary epithelial cells and macrophages. JGTW also demonstrates potency in reducing inflammatory cell recruitment and allergic response in the OVA-induced asthmatic mouse model, suggesting the clinical effectiveness of JGT on relieving symptoms of asthma. In addition, paeoniflorin and glycyrrhizin were abundantly present in JGTW, which might contribute to its anti-inflammatory effects.