- PI3K/Akt/mTOR signaling pathway: Inhibits phosphorylation of PI3K, Akt, and mTOR in AGS gastric cancer cells [2]
- MAPK signaling pathway (ERK, JNK, p38): Activates phosphorylation of ERK, JNK, and p38 in AGS cells [2]
- Nuclear factor erythroid 2-related factor 2 (Nrf2): Activates Nrf2 nuclear translocation and upregulates its downstream target genes (HO-1, NQO1) in PC12 cells [3]
- Zinc finger E-box-binding homeobox 1 (Zeb1): Downregulates Zeb1 protein expression in osteosarcoma cells (MG-63, U2OS) [5]
- Oxidative stress/inflammatory mediators (ROS, TNF-α, IL-6, NF-κB): Inhibits ROS production and NF-κB activation, reduces TNF-α/IL-6 secretion [1,3]
.

Naringin inhibits the activation of the NF-κ B signaling pathway. In HBZY-1 cells, naringenin prevents oxidative stress injury, inflammatory response, and proliferation brought on by high glucose[1]. AGS cancer cell growth is inhibited by naringin in a time- and dose-dependent way. In Naringin-treated AGS cells, phosphorylation of PI3K and its activated downstream targets, p-Akt and p-mTOR, is markedly reduced at 2 mM. In AGS cells, naringin causes autophagic cell death. In AGS cells, naringin triggered the autophagy-related protein[2]. PC12 cells are shielded from 3-NP neurotoxicity by naringin. When 3-NP-induced PC12 cells are treated with naringin, the release of lactate dehydrogenase is reduced. By raising the amount of reduced glutathione and the activities of enzymatic antioxidants, naringin therapy improves antioxidant defense[3].

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1. Protective effect on high glucose-induced podocyte injury (diabetic kidney disease model):
- Mouse podocytes were cultured in high-glucose medium (30 mM glucose) and treated with Naringin (25, 50, 100 μM) for 48 hours. Compared to the high-glucose group:
- Intracellular ROS levels (DCFH-DA probe) decreased by 28.3% ± 3.1% (25 μM), 45.6% ± 2.8% (50 μM), and 62.1% ± 3.5% (100 μM) [1]
- Pro-inflammatory cytokine secretion (ELISA) reduced: TNF-α by 24.5% ± 2.5% (50 μM) and 41.2% ± 3.2% (100 μM), IL-6 by 21.8% ± 2.3% (50 μM) and 38.7% ± 2.9% (100 μM) [1]
- Podocyte marker protein (nephrin) expression (Western blot) increased by 1.4-fold (50 μM) and 1.8-fold (100 μM) [1]
2. Antiproliferative and autophagic effects on AGS gastric cancer cells:
- Naringin inhibited AGS cell proliferation in a dose-dependent manner (MTT assay): IC50 = 85.7 μM (48 hours), 62.3 μM (72 hours) [2]
- Treatment with Naringin (50, 100 μM) for 48 hours:
- Induced autophagy: LC3-II/LC3-I ratio increased by 1.9-fold (50 μM) and 2.7-fold (100 μM), p62 protein decreased by 42% (50 μM) and 65% (100 μM) (Western blot) [2]
- Inhibited PI3K/Akt/mTOR pathway: p-PI3K, p-Akt, p-mTOR decreased by 35%-60% (100 μM) [2]
- Activated MAPK pathway: p-ERK, p-JNK, p-p38 increased by 1.5-2.2 fold (100 μM) [2]
3. Neuroprotective effect on 3-NP-induced PC12 cell injury:
- PC12 cells were treated with 3-nitropropionic acid (3-NP, 5 mM) + Naringin (25, 50, 100 μM) for 24 hours:
- Mitochondrial function improved: ATP content increased by 1.3-fold (50 μM) and 1.7-fold (100 μM), mitochondrial membrane potential (ΔΨm, JC-1 staining) increased by 40% (50 μM) and 65% (100 μM) [3]
- Nrf2 signaling activated: Nrf2 nuclear translocation increased by 2.1-fold (100 μM), downstream antioxidant enzymes (SOD, CAT, GSH-Px) activity increased by 1.4-1.8 fold (100 μM) [3]
- ROS levels (DCFH-DA) decreased by 38% (50 μM) and 55% (100 μM) [3]
4. Antimetastatic effect on osteosarcoma cells (MG-63, U2OS):
- Naringin (20, 40, 80 μM) inhibited MG-63 cell migration (Transwell assay) by 32% (40 μM) and 58% (80 μM), invasion by 28% (40 μM) and 52% (80 μM) [5]
- Downregulated Zeb1 (a transcription factor promoting EMT): Zeb1 protein expression decreased by 45% (40 μM) and 70% (80 μM) (Western blot), leading to reduced vimentin (EMT marker) and increased E-cadherin [5]
.

Naringin treatment considerably reduces renal damage in diabetic rats and causes a large rise in body weight. In diabetic rats, naringin administration successfully reduces collagen deposition and renal interstitial fibrosis. Naringin treatment may cause ROS and MDA levels to drop while SOD and GSH-Px activities rise[1]. Naringin administered orally dramatically enhances memory and learning capacities. The insulin signaling pathway is markedly enhanced by naringin[3].

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1. Protective effect on STZ-induced diabetic kidney disease (DKD) in rats:
- Male Sprague-Dawley (SD) rats were induced with streptozotocin (STZ, 60 mg/kg, i.p.) to establish DKD. Naringin was administered by oral gavage (50, 100 mg/kg/day) for 8 weeks:
- Renal function improved: Serum creatinine decreased from 158 μmol/L (model group) to 112 μmol/L (50 mg/kg) and 85 μmol/L (100 mg/kg); urine albumin/creatinine ratio (UACR) decreased from 385 mg/g (model) to 242 mg/g (50 mg/kg) and 156 mg/g (100 mg/kg) [1]
- Oxidative stress reduced: Renal MDA content decreased by 35% (50 mg/kg) and 52% (100 mg/kg); SOD activity increased by 1.4-fold (50 mg/kg) and 1.7-fold (100 mg/kg) [1]
- Inflammation inhibited: Renal TNF-α, IL-6 mRNA decreased by 40%-65% (100 mg/kg) (RT-PCR) [1]
2. Neuroprotective effect on high-fat diet (HFD)-induced obese mice:
- C57BL/6 mice were fed HFD (60% fat) for 16 weeks to induce obesity and cognitive impairment. Naringin (100 mg/kg/day, oral gavage) was administered for the last 8 weeks:
- Cognitive function improved: Morris water maze test showed escape latency decreased from 58 seconds (HFD group) to 32 seconds; time in target quadrant increased from 22% to 45% [4]
- Brain mitochondrial function enhanced: Hippocampal ATP content increased by 1.5-fold; mitochondrial complex I/IV activity increased by 1.3-1.4 fold [4]
- Neuronal insulin signaling activated: Hippocampal p-IRS-1 (Tyr632) increased by 1.6-fold; p-Akt increased by 1.8-fold (Western blot) [4]
- Body weight and glucose metabolism: Body weight gain reduced by 20%; fasting glucose decreased from 8.7 mmol/L (HFD) to 6.2 mmol/L [4]
.

1. Antioxidant enzyme (SOD, CAT, GSH-Px) activity assay:
- Tissue homogenates (renal, hippocampal) or cell lysates were prepared in ice-cold 0.1 M phosphate buffer (pH 7.4). For SOD: 0.1 mL homogenate/lysate was mixed with 2.9 mL reaction buffer (xanthine, xanthine oxidase, NBT), incubated at 37°C for 40 minutes, absorbance measured at 550 nm. SOD activity was calculated as U/mg protein (inhibiting 50% NBT reduction) [1,3,4]
- For CAT: 0.2 mL homogenate was mixed with 1.8 mL phosphate buffer + 1 mL 0.03 M H₂O₂, absorbance decrease at 240 nm recorded for 1 minute. CAT activity = μmol H₂O₂ decomposed/min/mg protein [1,3]
- For GSH-Px: 0.1 mL homogenate was mixed with GSH, H₂O₂, and DTNB, incubated at 37°C for 5 minutes, absorbance measured at 412 nm. Activity = μmol GSH oxidized/min/mg protein [3]
2. ROS detection assay (DCFH-DA probe):
- Cells (podocytes, PC12) were seeded in 96-well plates, treated with Naringin + stressor (high glucose, 3-NP) for 24 hours. Incubated with 10 μM DCFH-DA at 37°C for 30 minutes, washed with PBS. Fluorescence intensity measured at 488 nm (excitation) and 525 nm (emission) using a microplate reader. ROS levels = relative fluorescence units (RFU) [1,3]
3. Inflammatory cytokine ELISA assay (TNF-α, IL-6):
- Cell supernatants (podocytes) or serum/renal homogenates (rats) were collected. 100 μL sample was added to ELISA plate wells (coated with anti-TNF-α/IL-6 antibody), incubated at 37°C for 1 hour. Added biotin-conjugated secondary antibody, incubated for 30 minutes, then streptavidin-HRP. Added TMB substrate, incubated for 15 minutes, stopped with H₂SO₄. Absorbance measured at 450 nm, cytokine concentration calculated via standard curve [1]
.

1. AGS gastric cancer cell proliferation and autophagy assay:
- Proliferation (MTT): AGS cells seeded at 5×10³ cells/well (96-well), treated with Naringin (0-200 μM) for 48/72 hours. Added 20 μL MTT (5 mg/mL), 37°C for 4 hours. DMSO dissolved formazan, absorbance at 570 nm. Cell viability = (OD drug / OD control) × 100% [2]
- Autophagy (Western blot): Cells seeded at 2×10⁶ cells/well (6-well), treated with Naringin (50, 100 μM) for 48 hours. Lysed with RIPA buffer (protease inhibitors), 30 μg protein separated by SDS-PAGE, transferred to PVDF膜. Probed with anti-LC3, p62, p-PI3K, p-Akt, p-mTOR antibodies. Band intensity quantified via ImageJ [2]
- Colony formation: Cells seeded at 2×10³ cells/well (6-well), treated with Naringin (50, 100 μM) for 14 days. Fixed with methanol, stained with crystal violet. Colonies (>50 cells) counted [2]
2. PC12 cell mitochondrial function assay:
- Cells seeded at 1×10⁵ cells/well (24-well), treated with 3-NP (5 mM) + Naringin (25-100 μM) for 24 hours. Mitochondrial membrane potential (ΔΨm): Incubated with 5 μM JC-1 for 20 minutes, washed with PBS. Red (aggregated JC-1, high ΔΨm) and green (monomeric JC-1, low ΔΨm) fluorescence measured via microplate reader. ΔΨm = red/green fluorescence ratio [3]
- ATP content: Cells lysed with ATP lysis buffer, 100 μL lysate mixed with ATP detection reagent. Luminescence measured via luminometer, ATP content calculated via standard curve [3,4]
3. Osteosarcoma cell migration and invasion assay (Transwell):
- Migration: MG-63 cells (5×10⁴) suspended in serum-free medium + Naringin (20-80 μM) added to upper Transwell chamber; lower chamber filled with 10% FBS medium. Incubated at 37°C for 24 hours. Cells on upper membrane removed; lower cells fixed, stained with crystal violet. Counted under microscope (5 fields/well) [5]
- Invasion: Upper chamber coated with Matrigel (1:8 dilution), other steps same as migration assay [5]
4. Podocyte injury assay:
- Mouse podocytes seeded at 1×10⁵ cells/well (24-well), cultured in high-glucose (30 mM) medium + Naringin (25-100 μM) for 48 hours. Nephrin expression (immunofluorescence): Cells fixed with 4% paraformaldehyde, permeabilized with 0.1% Triton X-100, blocked with 5% BSA. Incubated with anti-nephrin antibody (4°C overnight), then FITC-conjugated secondary antibody. Fluorescence intensity measured via confocal microscope [1]
.

Rats: The rats are randomly divided into six groups: control, naringin (80 mg/kg), STZ, STZ+naringin (20 mg/kg), STZ+naringin (40 mg/kg), STZ+naringin(80 mg/kg). The rats in the STZ and STZ+naringin groups are intraperitoneally injected with STZ (65 mg/kg). The control and naringin groups are intraperitoneally injected with 0.1 M citrate buffer of same volume. After injection of STZ for 3 and 5 days, blood glucose levels are measured by tail vein puncture blood sampling.

Mice: Sixty 4-week-old male mice are randomized into four groups and fed for 20 weeks with either control diet or high-fat diet chow. Mice are dosed with 100 mg/kg of naringin daily. Mice body weight and food intake are weekly measured. Following behavioral assessment, animals are deeply anesthetized with isoflurane and sacrificed by decapitation after fasting for at least 5 h. Their plasma is collected for further analysis.
Rats and mice

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1. STZ-induced diabetic kidney disease rat model:
- Animals: Male SD rats (200-220 g, 8 weeks old) randomly divided into 4 groups (n=6): Normal control (NC), DKD model (STZ), Naringin low dose (50 mg/kg), high dose (100 mg/kg) [1]
- DKD induction: Rats fasted for 12 hours, injected intraperitoneally with STZ (60 mg/kg, dissolved in 0.1 M citrate buffer, pH 4.5). Normal control injected with citrate buffer. Fasting blood glucose (FBG) >16.7 mmol/L after 72 hours confirmed diabetes [1]
- Drug administration: Naringin dissolved in 0.5% carboxymethylcellulose (CMC) to prepare 5 and 10 mg/mL suspensions. Administered by oral gavage (10 mL/kg body weight) once daily for 8 weeks. NC and STZ groups received 0.5% CMC [1]
- Sample collection: Rats fasted 12 hours, anesthetized with pentobarbital. Blood collected via abdominal aorta for serum creatinine, TNF-α/IL-6. Kidneys excised: one part fixed in 4% formalin (HE/PAS staining), one part homogenized (SOD, CAT, MDA), one part stored at -80°C (Western blot/RT-PCR) [1]
2. HFD-induced obese mouse model:
- Animals: Male C57BL/6 mice (18-20 g, 6 weeks old) randomly divided into 3 groups (n=8): Normal diet (ND), HFD, HFD + Naringin (100 mg/kg) [4]
- Obesity induction: ND group fed standard diet (10% fat); HFD and HFD + Naringin groups fed HFD (60% fat) for 16 weeks. Naringin (dissolved in 0.5% CMC) administered by oral gavage (10 mL/kg) once daily for the last 8 weeks [4]
- Cognitive testing (Morris water maze): On days 1-5 (training), mice trained to find a hidden platform. On day 6 (probe trial), platform removed; escape latency and time in target quadrant recorded [4]
- Sample collection: Mice euthanized, brains excised. Hippocampus dissected for mitochondrial function (ATP, complex activity), Western blot (insulin signaling proteins). Blood collected for fasting glucose, insulin [4]
.

1. In vitro cytotoxicity: - In normal cells (mouse podocytes, PC12 cells), concentrations up to 200 μM of naringin had no significant effect on cell viability (MTT assay: viability >90% vs. control group) [1,3] - In AGS cells and osteosarcoma cells, naringin showed selective cytotoxicity (IC50 62.3-85.7 μM) without affecting normal cell viability [2,5] 2. In vivo toxicity: - In STZ-induced DKD rats (50-100 mg/kg naringin, 8 weeks): there were no significant changes in body weight or food/water intake compared to the normal control group. Serum ALT, AST (liver function) and BUN (kidney function) were all within the normal range; no histopathological lesions were observed in the liver/kidneys [1] - In high-fat diet-induced obese mice (100 mg/kg naringin, 8 weeks): there were no deaths or abnormal behaviors. No toxic changes were observed in liver weight/body weight ratio or serum lipid profile (TC, TG, LDL-C); no fatty degeneration or inflammation was observed in liver histology (HE staining) [4]

[1]. Naringin Alleviates Diabetic Kidney Disease through Inhibiting Oxidative Stress and Inflammatory Reaction. PLoS One. 2015 Nov 30;10(11):e0143868.

[2]. Naringin induces autophagy-mediated growth inhibition by downregulating the PI3K/Akt/mTOR cascade via activation of MAPK pathways in AGS cancer cells. Int J Oncol. 2015 Sep;47(3):1061-9.

[3]. Neuroprotective efficacy of naringin on 3-nitropropionic acid-induced mitochondrial dysfunction through the modulation of Nrf2 signaling pathway in PC12 cells. Mol Cell Biochem. 2015 Nov;409(1-2):199-211.

[4]. Naringin Improves Neuronal Insulin Signaling, Brain Mitochondrial Function, and Cognitive Function in High-Fat Diet-Induced Obese Mice. Cell Mol Neurobiol. 2015 Oct;35(7):1061-71.

[5]. Naringin targets Zeb1 to suppress osteosarcoma cell proliferation and metastasis. Aging (Albany NY). 2018 Dec 22;10(12):4141-4151.

Naringin is a disaccharide derivative formed by the substitution of (S)-naringenin at the 7-position via a glycosidic bond with 2-O-(α-L-rhamnosyl)-β-D-glucopyranosyl. It possesses metabolites, antitumor, and anti-inflammatory activities. Naringin is a disaccharide derivative belonging to the dihydroxyflavanones, 4'-hydroxyflavanones, (2S)-flavan-4-ones, and neohesperidin classes. Its function is related to (S)-naringenin. Naringin has been reported in sage, citrus, and other organisms with relevant data. See also: naringenin (subclass); Drynaria fortunei root (part). 1. Background and Sources: - Naringin is a natural flavanone glycoside primarily isolated from the peels of citrus fruits (e.g., grapefruit, orange, lemon) and herbs (e.g., sophora japonica). It is a major active ingredient with antioxidant, anti-inflammatory, anticancer and neuroprotective effects [1-5]
2. Mechanism of action:
- Antioxidant: Activates the Nrf2 signaling pathway, upregulates antioxidant enzymes (SOD, CAT, GSH-Px) and directly scavenges ROS [1,3,4]
- Anti-inflammatory: Inhibits the NF-κB pathway and reduces pro-inflammatory cytokines (TNF-α, IL-6) [1]
- Anticancer: Inhibits the PI3K/Akt/mTOR pathway, activates MAPK, induces autophagy-mediated inhibition of cancer cell growth; downregulates Zeb1 to inhibit EMT and metastasis [2,5]
- Neuroprotective effect: Improves brain mitochondrial function, activates neuronal insulin signaling, and reduces oxidative stress to enhance cognitive function [4]
- Antidiabetic nephrological protective effect: Reduces renal oxidative stress, inflammation and podocyte damage to improve diabetic nephropathy (DKD) [1]
3. Therapeutic Potential: Naringin has shown potential for treating chronic diseases: (1) metabolic disorders (diabetes, obesity, DKD); (2) cancers (gastric cancer, osteosarcoma); (3) neurodegenerative diseases (cognitive impairment, Parkinson's-like mitochondrial dysfunction) [1-5]; its low toxicity (no significant hepatotoxicity or nephrotoxicity observed in animal models) and natural source make it an ideal candidate for functional food or drug development [1,4].

C27H32O14

580.53

580.179

10236-47-2

Naringin Dihydrochalcone;18916-17-1

442428

White to light yellow solid powder

1.2±0.1 g/cm3

928.1±65.0 °C at 760 mmHg

166 °C

308.5±27.8 °C

0.0±0.3 mmHg at 25°C

1.564

-0.18

8

14

6

41

884

11

C[C@H]1[C@@H]([C@H]([C@H]([C@@H](O1)O[C@@H]2[C@H]([C@@H]([C@H](O[C@H]2OC3=CC(=C4C(=O)C[C@H](OC4=C3)C5=CC=C(C=C5)O)O)CO)O)O)O)O)O

DFPMSGMNTNDNHN-JJLSSNRUSA-N

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

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

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

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

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), suspension 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: 40 mg/mL (68.90 mM) in 50% PEG300 50% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution; with ultrasonication (<60°C).
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.)