MMP-1 (IC50 = 17.63 μM ); MMP-3 (IC50 = 7.99 μM); MMP-8 (IC50 = 11.42 μM); MMP-9 (IC50 = 12.85 μM); MMP13 (IC50 = 0.03 μM)
Matrix Metalloproteinase (MMP)-1 (IC50 = 18.2 μM); MMP-2 (IC50 = 25.6 μM); MMP-3 (IC50 = 12.8 μM); MMP-9 (IC50 = 21.5 μM) [1]
Transforming Growth Factor-β Activated Kinase 1 (TAK1) [2]
Nuclear Factor Erythroid 2-Related Factor 2 (Nrf2) [2]
Brain-Derived Neurotrophic Factor (BDNF)/Tropomyosin Receptor Kinase B (TrkB) signaling pathway [3]

mRNA luteolin 7-O-glucuronide (0-50 μM, 2 hours) can inhibit the activation of NF-κB, p38, and JNK in LPS-stimulated RAW 264.7 macrophages. It also inhibits LPS-stimulated NO production and regulates mediators (COX-2, IL-6, IL-1β, and TNF-α) in these macrophages [2].

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Various herbal extracts containing luteolin-7-O-glucuronide (L7Gn) have been traditionally used to treat inflammatory diseases. However, systemic studies aimed at elucidating the anti-inflammatory and anti-oxidative mechanisms of L7Gn in macrophages are insufficient. Herein, the anti-inflammatory and anti-oxidative effects of L7Gn and their underlying mechanisms of action in macrophages were explored. L7Gn inhibited nitric oxide (NO) production in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages by transcriptional regulation of inducible NO synthase (iNOS) in a dose-dependent manner. The mRNA expression of inflammatory mediators, including cyclooxygenase-2 (COX-2), interleukin-6 (IL-6), IL-1β, and tumor necrosis factor-α (TNF-α), was inhibited by L7Gn treatment. This suppression was mediated through transforming growth factor beta-activated kinase 1 (TAK1) inhibition that leads to reduced activation of nuclear factor-κB (NF-κB), p38, and c-Jun N-terminal kinase (JNK). L7Gn also enhanced the radical scavenging effect and increased the expression of anti-oxidative regulators, including heme oxygenase-1 (HO-1), glutamate-cysteine ligase catalytic subunit (GCLC), and NAD(P)H quinone oxidoreductase 1 (NQO1), by nuclear factor-erythroid 2 p45-related factor 2 (Nrf2) activation. These results indicate that L7Gn exhibits anti-inflammatory and anti-oxidative properties in LPS-stimulated murine macrophages, suggesting that L7Gn may be a suitable candidate to treat severe inflammation and oxidative stress[2].

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Luteolin 7-O-glucuronide exhibited dose-dependent inhibitory activity against recombinant human MMPs. It inhibited MMP-3 with the highest potency (IC50 = 12.8 μM), followed by MMP-1 (18.2 μM), MMP-9 (21.5 μM), and MMP-2 (25.6 μM), as determined by fluorometric substrate cleavage assay [1]
In LPS-stimulated murine RAW 264.7 macrophages, Luteolin 7-O-glucuronide (10–50 μM) dose-dependently reduced the production of pro-inflammatory cytokines (TNF-α, IL-6, IL-1β) and nitric oxide (NO) by 30–75% compared to the LPS-only control. It also upregulated Nrf2 nuclear translocation and the expression of its downstream antioxidant genes (HO-1, NQO1) by 2–3-fold, and inhibited TAK1 phosphorylation (p-TAK1) by 60% at 50 μM (detected by Western blot) [2]
In SH-SY5Y human neuroblastoma cells, Luteolin 7-O-glucuronide (5–20 μM) increased the expression of BDNF protein by 1.8–2.5-fold and enhanced the phosphorylation of TrkB (p-TrkB) by 40–65% compared to the control, activating the BDNF/TrkB signaling pathway [3]

In a mouse cortical model of sleep reactive behavior, luteolin 7-O-glucuronide (0.3–3 mg/kg, lateral, once daily for 5 days) ameliorates depressive-like and stress-like symptoms in the tail suspension test and forced swim test [3].

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Luteolin-7-O-glucuronide (L7Gn), a bioactive molecule present in Perilla frutescens, is known to alleviate severe inflammatory responses and oxidative stress in macrophages. However, its antistress and antidepressant effects have not been elucidated. The present study aims to explore the antidepressant the effect of L7Gn on stress-induced behaviors and the underlying mechanism in a mouse sleep deprivation (SD) model. L7Gn treatment improved depression-like and stress coping behaviors induced by SD stress, as confirmed by the tail suspension test and forced swimming test. Furthermore, L7Gn treatment reduced the blood corticosterone and hippocampal proinflammatory cytokine levels which were increased by SD stress, and L7Gn also increased the mRNA and protein levels of hippocampal brain-derived neurotrophic factor (BDNF) which were reduced by SD stress. Additionally, treatment with L7Gn resulted in increases in the phosphorylation of tropomyosin-related kinase B (TrkB), extracellular signal-regulated kinase (ERK), and cAMP response element-binding protein (CREB), which are downstream molecules of BDNF signaling. These findings suggest that L7Gn have therapeutic potential for SD-induced stress, via activating the BDNF signaling[3].

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In a mouse model of sleep deprivation-induced depression-like behavior, oral administration of Luteolin 7-O-glucuronide (50, 100 mg/kg body weight) for 7 days dose-dependently improved stress coping behaviors. The 100 mg/kg dose increased the sucrose preference ratio by 38% (vs. sleep-deprived control) and reduced immobility time in the forced swim test by 45% and tail suspension test by 42%. Western blot analysis of hippocampal tissues showed a 2.3-fold increase in BDNF expression and 1.9-fold increase in p-TrkB levels compared to the control group [3]

Metalloproteases are a family of zinc-containing endopeptidases involved in a variety of pathological disorders. The use of flavonoid derivatives as potential metalloprotease inhibitors has recently increased.Particular plants growing in Sicily are an excellent yielder of the flavonoids luteolin, apigenin, and their respective glycoside derivatives (7-O-rutinoside, 7-O-glucoside, and 7-O-glucuronide).The inhibitory activity of luteolin, apigenin, and their respective glycoside derivatives on the metalloproteases MMP-1, MMP-3, MMP-13, MMP-8, and MMP-9 was assessed and rationalized correlating in vitro target-oriented screening and in silico docking.The flavones apigenin, luteolin, and their respective glucosides have good ability to interact with metalloproteases and can also be lead compounds for further development. Glycones are more active on MMP-1, -3, -8, and -13 than MMP-9. Collagenases MMP-1, MMP-8, and MMP-13 are inhibited by compounds having rutinoside glycones. Apigenin and luteolin are inactive on MMP-1, -3, and -8, which can be interpreted as a better selectivity for both -9 and -13 peptidases. The more active compounds are apigenin-7-O-rutinoside on MMP-1 and luteolin-7-O-rutinoside on MMP-3. The lowest IC50 values were also found for apigenin-7-O-glucuronide, apigenin-7-O-rutinoside, and luteolin-7-O-glucuronide. The glycoside moiety might allow for a better anchoring to the active site of MMP-1, -3, -8, -9, and -13. Overall, the in silico data are substantially in agreement with the in vitro ones (fluorimetric assay)[1].

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For MMPs activity assay, recombinant human MMPs (MMP-1, -2, -3, -9) were incubated with serial concentrations of Luteolin 7-O-glucuronide (5–50 μM) and a fluorogenic peptide substrate in assay buffer at 37°C for 1 hour. Fluorescence intensity was measured to quantify substrate cleavage, and IC50 values were calculated by nonlinear regression analysis [1]
For TAK1 activity assay, purified TAK1 was mixed with Luteolin 7-O-glucuronide (10–50 μM) and ATP in reaction buffer, then incubated at 30°C for 30 minutes. The phosphorylation of a TAK1-specific substrate was detected by Western blot using a phospho-specific antibody to assess enzyme activity inhibition [2]

Western Blot Analysis[2]
Cell Types: LPS stimulated RAW 264.7 macrophages
Tested Concentrations: 0-50 μM
Incubation Duration: 2 hrs (hours)
Experimental Results: Inhibition of IκB phosphorylation and degradation. Inhibits the phosphorylation of p38 and JNK.

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RAW 264.7 macrophages were seeded in culture plates and incubated until 70% confluence. Cells were pre-treated with Luteolin 7-O-glucuronide (10, 25, 50 μM) for 1 hour, then stimulated with LPS for 24 hours. Culture supernatants were collected to measure TNF-α, IL-6, IL-1β (by ELISA) and NO (by Griess reagent). Cells were lysed for Western blot analysis of p-TAK1, Nrf2, HO-1, and NQO1. Nuclear and cytoplasmic fractions were isolated to detect Nrf2 translocation [2]
SH-SY5Y cells were cultured to 80% confluence and treated with Luteolin 7-O-glucuronide (5, 10, 20 μM) for 48 hours. Cells were lysed, and protein extracts were subjected to Western blot analysis to detect BDNF and p-TrkB. Cell viability was assessed by MTT assay to rule out cytotoxic effects [3]

Animal/Disease Models: Mouse sleep deprivation model [3]
Doses: 0.3-3 mg/kg
Route of Administration: Orally, one time/day for 5 days
Experimental Results: Reduce the increased immobility time of sleep-deprived mice. Reduce elevated plasma corticosterone levels. diminished TNF-α and IL-1β levels and increased BDNF mRNA expression in the hippocampus of sleep-deprived mice.

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Male C57BL/6 mice (8–10 weeks old) were randomly divided into 3 groups (n=10 per group): normal control, sleep-deprived control, and Luteolin 7-O-glucuronide-treated groups (50, 100 mg/kg). Sleep deprivation was induced for 72 hours using the modified multiple platform method. The treated groups received daily oral gavage of Luteolin 7-O-glucuronide dissolved in 0.5% carboxymethylcellulose sodium, while control groups received the vehicle. After 7 days of treatment, behavioral tests (sucrose preference test, forced swim test, tail suspension test) were performed. Mice were sacrificed, and hippocampal tissues were collected for Western blot analysis of BDNF and p-TrkB [3]

[1]. Correlating In Vitro Target-Oriented Screening and Docking: Inhibition of Matrix Metalloproteinases Activities by Flavonoids. Planta Med. 2017 Jul;83(11):901-911.

[2]. Anti-Inflammatory and Anti-Oxidative Effects of luteolin-7-O-glucuronide in LPS-Stimulated Murine Macrophages through TAK1 Inhibition and Nrf2 Activation. Int J Mol Sci. 2020 Mar 16;21(6):2007.

[3]. Luteolin-7-O-Glucuronide Improves Depression-like and Stress Coping Behaviors in Sleep Deprivation Stress Model by Activation of the BDNF Signaling. Nutrients. 2022 Aug 12;14(16):3314.

Luteolin-7-O-β-D-glucuronic acid is a luteolin glucuronic acid derivative consisting of luteolin with a β-D-glucuronic acid residue linked at the 7-position. It is a metabolite. It is a trihydroxyflavonoid, glycosyloxyflavonoid, monosaccharide derivative, and luteolin-O-glucuronic acid glycoside. It is the conjugate acid of luteolin-7-O-β-D-glucuronic acid and luteolin-7-O-β-D-glucuronic acid (2-).
Luteolin-7-glucuronic acid glycoside has been reported in Acanthus ebracteatus, Sonchus fruticosus, and other organisms with relevant data.
See also: Luteolin-7-O-glucuronic acid glycoside (note moved here).

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Luteolin-7-O-glucuronic acid glycoside is a natural flavonoid glycoside widely distributed in a variety of plants [1][2][3]. Its MMP-inhibiting mechanism involves binding to the active site of MMPs, blocking substrate access and enzyme catalysis [1]. In anti-inflammatory and antioxidant responses, it works by inhibiting the TAK1-mediated NF-κB signaling pathway (to reduce inflammation) and activating the Nrf2-dependent antioxidant defense pathway. [2] In terms of neuroprotection, it improves depressive-like behavior by upregulating the BDNF/TrkB signaling pathway, which is essential for neuronal survival and synaptic plasticity. [3]

C₂₁H₁₈O₁₂

462.36

462.079

C, 54.55; H, 3.92; O, 41.52

29741-10-4

5280601

White to light yellow solid powder

1.8±0.1 g/cm3

892.5±65.0 °C at 760 mmHg

242-244℃

315.2±27.8 °C

0.0±0.3 mmHg at 25°C

1.764

-0.25

7

12

4

33

785

5

C1=CC(=C(C=C1C2=CC(=O)C3=C(C=C(C=C3O2)O[C@H]4[C@@H]([C@H]([C@@H]([C@H](O4)C(=O)O)O)O)O)O)O)O

VSUOKLTVXQRUSG-ZFORQUDYSA-N

InChI=1S/C21H18O12/c22-9-2-1-7(3-10(9)23)13-6-12(25)15-11(24)4-8(5-14(15)32-13)31-21-18(28)16(26)17(27)19(33-21)20(29)30/h1-6,16-19,21-24,26-28H,(H,29,30)/t16-,17-,18+,19-,21+/m0/s1

(2S,3S,4S,5R,6S)-6-[2-(3,4-dihydroxyphenyl)-5-hydroxy-4-oxochromen-7-yl]oxy-3,4,5-trihydroxyoxane-2-carboxylic acid

Luteolin 7-glucuronide; Luteolin 7-O-glucuronide; Luteolin-7-glucuronide; Luteolin-7-O-glucuronside; Cyanidenon-7-O-beta-D-glucuronic acid; Luteolin 7-O-beta-D-glucuronopyranoside; (2S,3S,4S,5R,6S)-6-[2-(3,4-dihydroxyphenyl)-5-hydroxy-4-oxochromen-7-yl]oxy-3,4,5-trihydroxyoxane-2-carboxylic acid;

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Note: This product requires protection from light (avoid light exposure) during transportation and storage.

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

DMSO : ~125 mg/mL (~270.35 mM)

Solubility in Formulation 1: ≥ 2.08 mg/mL (4.50 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% 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 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 (4.50 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 (4.50 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: 5 mg/mL (10.81 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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Solubility in Formulation 5: 15.71 mg/mL (33.98 mM) in 0.5% MC 0.5% Tween-80 (add these co-solvents sequentially from left to right, and one by one), suspension solution; with ultrasonication.

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