## 4 Discussion  
The prevalence of allergic diseases and conditions, including allergic rhinitis, atopic dermatitis, and asthma, is increasing worldwide, posing a significant economic burden on affected individuals [66; 67]. Several polyphenols are known to possess anti-allergic activities by inhibiting mast cell activation in vitro. For example, tricin (100-500 \(\upmu\)g/mL) inhibits the phosphorylation of protein kinase B (Akt), p38, ERK, and JNK in activated RBL-2H3 mast cells, as well as Syk, Lyn, PLCy, and PKC8 [54]. Similarly, saponarin (80 \(\upmu\)M) inhibits the expression of FccRI\(\alpha\)/\(\gamma\) in IgE-activated RBL-2H3 mast cells and suppresses mast cell activation by reducing the phosphorylation of Syk, PLCy, ERK, JNK, and p38 [55]. Quercetin inhibits mast cell activation by reducing the surface expression of Fc\(\epsilon\)RI and cytokine production (IL-6, IL-33) in IgE-activated BMMC (bone marrow-derived mast cells) [53]. _Lonicera caerulea_ fruit is a rich source of C3G. C3G has been shown to exert anti-allergic effects through regulation of IL-1R\(\alpha\)-STAT6 signaling, strengthening of the intestinal epithelial barrier, and regulation of Th1/Th2 immune balance in the OVA-induced asthma mice model [40; 41]. Additionally, anti-DNP inhibited \(\beta\)-hexosaminidase and histamine secretion from IgE-stimulated RBL-2H3 mast cells and suppressed passive cutaneous anaphylaxis in IgE-sensitized mice [42]. Despite these anti-allergic properties of C3G, studies specifically focusing on the anti-allergic effects of _Lonicera caerulea_ fruit have not yet been conducted. In this study, we evaluated the potential anti-allergic effects of HR2303-30E and C3G. We assessed its impact on the secretion of \(\beta\)-hexosaminidase and histamine in RBL-2H3 mast cells. Additionally, we investigated the modulation of key signaling pathways, including FccRI, Syk, Fyn, Src, PLCy1, PKC8, and MAPK. _Lonicera caerulea_ fruit has various components, such as ascorbic acid, anthocyanins, flavonoids, and phenolic acids, and exhibits antioxidant activity [68; 69]. We measured the total phenolic content, total flavonoid content, and antioxidant activity of HR2302-30E in this study. First, we measured the phenolic content of _Lonicera caerulea_ berries. Total phenolic and flavonoid contents of HR2302-30E were 18.73 \(\pm\) 0.74 mg GAE/g and \(11.83\pm 0.81\) mg QE/g, respectively, which are higher than those reported in previous studies (4.22-11.11 mg GAE/g and 4.36-11.37 mg QE/g, respectively) [70; 71; 72]. Antioxidant activity cannot be tested using a single method and must be analyzed using various procedures [57]. Here, the antioxidant activity of HR2302-30E was measured using four methods: DPPH, ABTS, FRAP, and ORAC assors. DPPH radical scavenging assay is based on the electron donation of antioxidants to neutralize DPPH, a stable chromogenic radical with a deep purple color [73]. The ABTS test is based on a decolorization reaction, in which a stable radical cation (ABTS \({}^{\bullet+}\)), a blue-green-emitting group, is removed by antioxidants either through direct reduction via electron donation or radical quenching via hydrogen atom donation [74]. The FRAP method measures the potential of a reducing agent (antioxidant) to reduce ferric ion (Fe\({}^{3+}\))-ligand to blue ferrous complex (Fe\({}^{2+}\)) [75]. The ORAC assay measures the radical chain scavenging activity of antioxidants by monitoring the inhibition of peroxyl radical oxidation [76]. DPPH radical scavenging activity (IC50), ABTS radical scavenging activity (IC50), FRAP activity, and ORAC value of HR2302-30E were 3.40 mg/mL, 5.56 mg/mL, 93.84 mmol TE/g, and 181.59 \(\upmu\)mol TE/g, respectively. In a previous study, the antioxidant activity of _Lonicera caerulea_ fruit varied from 5.83 to 7.8 mg/mL in the DPPH assay, from 27.96 to 46.9 \(\upmu\)M TE/g in the FRAP assay, and from 267.19 to 262.44 \(\upmu\)M TE/g in the ORAC assay [70]. These differences in results may be due to the different cultivars, cultivation locations, and harvest periods.