Inhibition of phosphorylation of Syk, LAT, and PLCγ1 signaling pathway transduction; inhibition of Ca2+ influx; suppression of phosphorylation of MAPK signaling pathways (p38, ERK, JNK); suppression of NF-κB signaling pathway (IκBα, IKKα/β, p65). [3]

RBL-2H3 cells remain viable in the presence of narirutin (200 μM, 20 h). In RBL-2H3 cells, narirutin (25 μM, 2.5 h) inhibits the release of β-hex, histamine, IL-4, and TNF-a, with levels below 40%. [3].

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Narirutin (0-200 μM) showed no significant effect on RBL-2H3 cell viability. [3]
- Narirutin inhibited RBL-2H3 cell degranulation as measured by β-hexosaminidase (β-hex) release. At concentrations of 25.00, 50.00, and 100.00 μM, β-hex release was below 40%. [3]
- Narirutin significantly reduced histamine release in IgE-sensitized RBL-2H3 cells. At concentrations of 6.25, 12.50, 25.00, 50.00, and 100.00 μM, histamine levels were 53.80±0.77, 52.53±2.27, 48.09±2.72, 48.80±3.19, and 48.37±2.12 ng/mL respectively (compared to sensitized group). [3]
- Narirutin significantly decreased IL-4 production. At concentrations of 12.50, 25.00, 50.00, and 100.00 μM, IL-4 levels were 62.76±0.91, 58.93±0.66, 54.55±1.28, and 51.63±1.58 ng/mL respectively. [3]
- Narirutin significantly decreased TNF-α release. At concentrations of 12.50, 25.00, 50.00, and 100.00 μM, TNF-α levels were 258.1±23.26, 217.1±10.15, 201.1±30.51, and 223.1±31.43 pg/mL respectively. [3]
- Atomic force microscopy (AFM) revealed that narirutin (100.00 μM) stabilized the morphology of activated RBL-2H3 cells, reduced the appearance of holes on the cell membrane, and suppressed the transfer of vesicles to the cell edge and supernatant induced by DNP-BSA stimulation. [3]
- Narirutin (25.00-100.00 μM) significantly inhibited Ca2+ influx in RBL-2H3 cells, with mean intracellular fluorescence intensity significantly weaker than the sensitized group; the 100.00 μM group showed similar intensity to the control group. [3]
- Narirutin had no effect on mRNA expression of FcεRI α, β, or γ subunits in RBL-2H3 cells. [3]
- Western blot analysis showed that narirutin suppressed phosphorylation of ERK1/2 and JNK in a dose-dependent manner, and suppressed p38 phosphorylation in a dose-independent manner in IgE-sensitized RBL-2H3 cells. [3]
- Narirutin suppressed phosphorylation of IκBα in a dose-dependent manner (40-100 μM) and suppressed IKKα/β phosphorylation in a dose-independent manner. Expression of phosphorylated p65 was significantly increased by narirutin at 40.00, 60.00, and 80.00 μM compared to sensitized group. [3]
- Narirutin decreased expression of P-Syk, P-LAT, and P-PLCγ1, but had no significant effect on expression of P-Lyn. [3]

Mice challenged with OVA have lower peripheral blood eosinophil counts when given nerirutin (10 mg/kg, p.o.) The OVA-induced increase in IL-4 is blocked by narirutin (10 mg/kg, p.o. ), but the increase in IL-5 is unaffected [2].

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Oral administration of narirutin at 10 mg/kg (but not 0.1 or 1 mg/kg) significantly reduced ovalbumin (OVA)-induced airway inflammation in NC/Nga mice. It diminished infiltration of inflammatory cells (mainly eosinophils) and mucus-producing cells in lung tissue, decreased eosinophil counts in peripheral blood and bronchoalveolar lavage fluid (BALF), reduced interleukin (IL)-4 levels in BALF, and lowered serum IgE levels. No significant effect on OVA-induced increase of IL-5 in BALF was observed. [2]

Histological analysis showed OVA-induced inflammation scores were significantly reduced by 10 mg/kg narirutin. In H&E-stained sections, the score decreased from 2.24±0.23 (OVA control) to 1.49±0.40; in PAS-stained sections, the score decreased from 2.43±0.53 (OVA control) to 1.88±0.48. [2]

Cell viability assay: RBL-2H3 cells were cultured in 96-well plates at 1.0×10^4 cells/well for 24 h, then treated with different concentrations of narirutin (0-200 μM) for 20 h. Cell viability was analyzed using CCK-8 kits. [3]
- β-Hexosaminidase release assay: RBL-2H3 cells (1×10^5 cells/well) were sensitized with 0.5 μg/mL anti-DNP monoclonal mouse IgE overnight, then treated with narirutin (0-200 μM) for 2.5 h, followed by stimulation with 0.5 μg/mL DNP-BSA for 1 h. The reaction was stopped on ice for 10 min. Supernatants were collected, centrifuged (4°C, 14,800 rpm, 10 min), and incubated with 50 μL of 4-nitrophenyl N-acetyl-β-D-glucosaminide (1 mM) for 1.5 h at 37°C. The reaction was stopped with 200 μL/well stop solution (0.1 mM Na2CO3/NaHCO3). OD values at 405 nm were measured. [3]
- Cytokine measurement: RBL-2H3 cells (5×10^5 cells/well) in 6-well plates were sensitized with 0.5 μg/mL anti-IgE, treated with narirutin (0-100 μM) for 2.5 h, then stimulated with 0.5 μg/mL DNP-BSA for 1 h. Reactions were stopped by ice-bath. Cell supernatants were collected, centrifuged at 4°C, 2500 rpm for 20 min, and cytokines (IL-4, histamine, TNF-α) were measured by ELISA kits. [3]
- AFM imaging: RBL-2H3 cells (1×10^5 cells/well) on cover slides in 6-well plates were sensitized with 0.5 μg/mL anti-DNP monoclonal mouse IgE for 12 h, washed with PBS, treated with narirutin (100.00 μM) for 2.5 h, then stimulated with 0.5 μg/mL DNP-BSA. Cells were washed with PBS, fixed with 4% paraformaldehyde for 10 min, washed with distilled water, dried in air, and observed by atomic force microscopy. [3]
- Intracellular Ca2+ concentration measurement: RBL-2H3 cells (1×10^5 cells/well) in 96-well black plates were sensitized with 0.5 μg/mL anti-DNP monoclonal mouse IgE for 12 h, washed with D-PBS, incubated with Fluo 3-AM (5 μM) for 40 min, then treated with narirutin (0.00, 40.00, 60.00, 80.00, 100.00 μM) for 2.5 h, and finally stimulated with 0.5 μg/mL DNP-BSA. Fluorescence intensity was measured under an inverted fluorescence microscope, and mean fluorescence intensity was calculated using Image J. [3]
- mRNA expression measurement: RBL-2H3 cells (1×10^4 cells/plate) were treated with narirutin (0.00, 40.00, 60.00, 80.00, 100.00 μM). mRNA was isolated using a magnetic bead-based kit, reverse transcribed, and PCR performed using SYBR qPCR SuperMix. Primer pairs for FcεRIα, FcεRIβ, FcεRIγ, and GAPDH were used. Three-step PCR amplification: 95°C for 1 min, then 95°C for 20 s, 60°C for 20 s, 72°C for 30 s for 40 cycles. Products were analyzed by 1.5% agarose gel electrophoresis and visualized. [3]
- Immunoblot analysis: RBL-2H3 cells (1×10^6 cells/plate) were treated as described. After DNP-BSA stimulation, proteins were extracted with RIPA lysis buffer. Lysates were centrifuged at 14,800 rpm for 10 min at 4°C. Protein concentrations were measured. 50 μg of supernatant protein was analyzed by SDS-PAGE and transferred to PVDF membranes. Membranes were blocked with skim milk or bovine albumin for 2-4 h, incubated with primary antibodies at 4°C overnight, then with secondary antibodies for 1 h at room temperature. Immunoblotting detection was performed using chemiluminescent gel imager. [3]

Animal Model: Allergic asthma female NC/Nga mice model[2]
Dosage: 0.1, 1 or 10 mg/kg
Administration: Orally, 7d
Result: Reduced eosinophil counts in peripheral blood. Blocked the OVA-induced increase in IL-4.

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Female NC/Nga mice (6-8 weeks old) were used. Mice were immunized intraperitoneally on Day 0 and Day 7 with 100 μg ovalbumin (OVA) plus 1.6 mg aluminum hydroxide in 200 μL sterile saline. Non-sensitized controls received only aluminum hydroxide in saline. On Days 14, 15, and 16, sensitized mice were exposed to aerosolized OVA (10 mg/mL in saline) for 10 min each day; non-sensitized mice were exposed to saline only. Narirutin was administered orally at doses of 0.1, 1, or 10 mg/kg body weight in sterile water (20 μL volume per mouse) daily from Day 7 to Day 16. Control groups received sterile water only. Mice were sacrificed 24 h after the last aerosol challenge (Day 17). [2]

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Female NC/Nga mice (6-8 weeks old) were used. Mice were immunized intraperitoneally on Day 0 and Day 7 with 100 μg ovalbumin (OVA) plus 1.6 mg aluminum hydroxide in 200 μL sterile saline. Non-sensitized controls received only aluminum hydroxide in saline. On Days 14, 15, and 16, sensitized mice were exposed to aerosolized OVA (10 mg/mL in saline) for 10 min each day; non-sensitized mice were exposed to saline only. Narirutin was administered orally at doses of 0.1, 1, or 10 mg/kg body weight in sterile water (20 μL volume per mouse) daily from Day 7 to Day 16. Control groups received sterile water only. Mice were sacrificed 24 h after the last aerosol challenge (Day 17). [2]

[1]. Structure-based Discovery of Narirutin as a Shikimate Kinase Inhibitor with Anti-tubercular Potency.Curr Comput Aided Drug Des. 2019 Oct 25.

[2]. Narirutin inhibits airway inflammation in an allergic mouse model.Clin Exp Pharmacol Physiol. 2007 Aug;34(8):766-70.

[3]. Inhibitory activity of narirutin on RBL-2H3 cells degranulation. Immunopharmacol Immunotoxicol. 2021, 43, 1.

Naringin is a disaccharide derivative formed by the substitution of (S)-naringenin at the 7-position via a glycosidic bond with a 6-O-(6-deoxy-α-L-mannopyranosyl)-β-D-glucopyranosyl group. It possesses anti-inflammatory, antioxidant, and metabolic activities. Naringin is a disaccharide derivative belonging to the dihydroxyflavanone, 4'-hydroxyflavanone, (2S)-flavan-4-one, and rutin glycoside classes. Its function is related to (S)-naringenin. Naringin has been reported in citrus fruits (Citrus sulcata), citrus fruits (Citrus reticulata), and other organisms with relevant data. See also: Orange peel (partial).

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Narirutin (naringenin-7-O-β-D-rutinoside) is a flavonoid abundant in citrus fruits such as grapefruits and oranges. It has anti-allergic properties and has been shown to inhibit histamine release from rat peritoneal mast cells in vitro and exert inhibitory effect on allergic skin reactions in a murine model of atopic dermatitis (according to previous studies). In the present study, the anti-inflammatory effect of narirutin in allergic airway inflammation is likely associated with reduction of OVA-induced increases of IL-4 and IgE, rather than IL-5. The findings suggest narirutin may be effective in treatment of bronchial asthma. [2]

C27H32O14

580.539

580.179

C, 55.86; H, 5.56; O, 38.58

14259-46-2

Isonaringin;108815-81-2

442431

Solid powder

1.7±0.1 g/cm3

924.3±65.0 °C at 760 mmHg

152-190ºC

307.3±27.8 °C

0.0±0.3 mmHg at 25°C

1.708

2.07

8

14

6

41

884

11

O1[C@]([H])([C@@]([H])([C@]([H])([C@@]([H])([C@@]1([H])C([H])([H])O[C@@]1([H])[C@@]([H])([C@@]([H])([C@]([H])([C@]([H])(C([H])([H])[H])O1)O[H])O[H])O[H])O[H])O[H])O[H])OC1=C([H])C(=C2C(C([H])([H])[C@@]([H])(C3C([H])=C([H])C(=C([H])C=3[H])O[H])OC2=C1[H])=O)O[H]

HXTFHSYLYXVTHC-AJHDJQPGSA-N

InChI=1S/C27H32O14/c1-10-20(31)22(33)24(35)26(38-10)37-9-18-21(32)23(34)25(36)27(41-18)39-13-6-14(29)19-15(30)8-16(40-17(19)7-13)11-2-4-12(28)5-3-11/h2-7,10,16,18,20-29,31-36H,8-9H2,1H3/t10-,16-,18+,20-,21+,22+,23-,24+,25+,26+,27+/m0/s1

(S)-5-hydroxy-2-(4-hydroxyphenyl)-7-(((2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-((((2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6-methyltetrahydro-2H-pyran-2-yl)oxy)methyl)tetrahydro-2H-pyran-2-yl)oxy)chroman-4-one

Naringenin 7-beta-rutinoside, Naringenin 7-O-rutinoside, (2S)-Narirutin;Isonaringenin, Isonaringin

2934.99.03.00

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Note: Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture and light.

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

DMSO : 100~125 mg/mL ( 172.25~215.32 mM)

Solubility in Formulation 1: 2.08 mg/mL (3.58 mM) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution; with sonication.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL.
Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution.

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Solubility in Formulation 2: ≥ 2.08 mg/mL (3.58 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly.
Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution.

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Solubility in Formulation 3: ≥ 2.08 mg/mL (3.58 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.

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Solubility in Formulation 4: 10% DMSO+40% PEG300+5% Tween-80+45% Saline: 2.08 mg/mL (3.58 mM)

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Solubility in Formulation 5: 10 mg/mL (17.23 mM) in 50% PEG300 50% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution; with ultrasonication.
Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution.

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 (Please use freshly prepared in vivo formulations for optimal results.)