NF-κB

1. For collagen-induced arthritis (CIA) study: Splenocytes were isolated from CIA mice, adjusted to a concentration of 2×10^6 cells/mL, and seeded in culture plates. The cells were stimulated with lipopolysaccharide (LPS) and treated with different concentrations of Eleutheroside E (1 μM, 5 μM, 10 μM) for 24 hours. The supernatant was collected, and enzyme-linked immunosorbent assay (ELISA) was used to detect the levels of inflammatory cytokines. The results showed that Eleutheroside E reduced the secretion of interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), and interleukin-6 (IL-6) in a concentration-dependent manner [1]
2. For type 2 diabetes study: 3T3-L1 preadipocytes were induced to differentiate into mature adipocytes. The differentiated adipocytes were treated with different concentrations of Eleutheroside E (2.5 μM, 5 μM, 10 μM) and insulin for 24 hours. Glucose uptake assay was performed to measure glucose transport activity mediated by glucose transporter 4 (GLUT4), and Western blot analysis was used to detect the phosphorylation levels of insulin receptor substrate-1 (IRS-1) and protein kinase B (AKT). The results indicated that Eleutheroside E enhanced insulin-induced glucose uptake and increased the phosphorylation of IRS-1 and AKT [2]

1. CIA mouse model experiment: CIA was induced in mice by immunization with bovine type II collagen emulsified in complete Freund's adjuvant. Mice were divided into 5 groups: control group, CIA model group, Eleutheroside E 10 mg/kg group, Eleutheroside E 20 mg/kg group, Eleutheroside E 40 mg/kg group. Eleutheroside E was administered via intraperitoneal injection once daily from day 21 to day 41 after the first immunization (total 21 days). Arthritis severity was scored weekly based on paw swelling and joint redness. At the end of the experiment, serum, joint tissues, and spleens were collected. The results showed that Eleutheroside E significantly reduced the arthritis score in a dose-dependent manner, alleviated pathological damage of joint tissues (reduced synovial hyperplasia and inflammatory cell infiltration), decreased serum levels of IL-1β, TNF-α, and IL-6, and lowered the spleen index (a marker of systemic inflammation) [1]
2. db/db mouse model experiment: Male db/db mice (a model of type 2 diabetes) were divided into 5 groups: control group, db/db model group, Eleutheroside E 5 mg/kg group, Eleutheroside E 10 mg/kg group, Eleutheroside E 20 mg/kg group. Eleutheroside E was administered via oral gavage once daily for 4 weeks. Body weight and fasting blood glucose (FBG) were measured weekly. At the end of the experiment, fasting insulin level was detected, and homeostatic model assessment of insulin resistance (HOMA-IR) was calculated. Glucose tolerance test (GTT) was performed to evaluate glucose metabolism. Adipose and liver tissues were collected for Western blot analysis. The results demonstrated that Eleutheroside E significantly reduced FBG, fasting insulin level, and HOMA-IR, improved glucose tolerance in GTT, and increased the phosphorylation levels of IRS-1 and AKT in adipose tissues [2]

1. AKT kinase activity assay (for type 2 diabetes study): Total protein was extracted from adipose tissues of db/db mice. The protein extract was incubated with AKT antibody-conjugated beads to immunoprecipitate AKT. The immunoprecipitated complex was mixed with kinase buffer, ATP, and AKT-specific substrate. The reaction was carried out at 37°C for 30 minutes, and the phosphorylation level of the substrate was detected by Western blot using phospho-substrate antibody. The activity of AKT was quantified based on the intensity of the phospho-substrate band. The results showed that Eleutheroside E increased AKT kinase activity in adipose tissues of db/db mice compared with the model group [2]

1. Splenocyte culture assay (for CIA study): Spleens were removed from CIA mice under sterile conditions and minced into small pieces. The tissue was digested with collagenase for 15 minutes, and then passed through a cell strainer to obtain a single-cell suspension. Red blood cells were lysed with lysing buffer, and the remaining cells were washed with culture medium. The splenocytes were resuspended in RPMI 1640 medium containing fetal bovine serum, penicillin, and streptomycin, adjusted to a concentration of 2×10^6 cells/mL, and seeded into 24-well plates (1 mL per well). LPS (1 μg/mL) was added to stimulate cytokine production, and Eleutheroside E was added at final concentrations of 1 μM, 5 μM, and 10 μM. The plates were incubated at 37°C in a 5% CO2 incubator for 24 hours. After incubation, the culture supernatant was collected by centrifugation (1000×g for 10 minutes) and stored at -80°C until ELISA detection of IL-1β, TNF-α, and IL-6 [1]
2. 3T3-L1 adipocyte differentiation and glucose uptake assay (for type 2 diabetes study): 3T3-L1 preadipocytes were cultured in DMEM medium containing fetal bovine serum until confluence. Two days after confluence (day 0), differentiation was induced by adding differentiation medium (DMEM with fetal bovine serum, isobutylmethylxanthine, dexamethasone, and insulin). On day 3, the medium was replaced with maintenance medium (DMEM with fetal bovine serum and insulin). On day 7, the cells were fully differentiated into adipocytes. For glucose uptake assay, the differentiated adipocytes were serum-starved for 4 hours, then treated with Eleutheroside E (2.5 μM, 5 μM, 10 μM) for 1 hour, followed by insulin (100 nM) for 30 minutes. 2-Deoxy-D-glucose (2-DG) was added to the medium, and the cells were incubated for another 20 minutes. The cells were washed with cold PBS, and the intracellular radioactivity of 2-DG was measured using a scintillation counter to evaluate glucose uptake [2]

1. CIA mouse model protocol: Female DBA/1 mice (6-8 weeks old) were used. On day 0, each mouse was immunized subcutaneously at the base of the tail with 100 μL of emulsion containing 100 μg bovine type II collagen and complete Freund's adjuvant. On day 21, a booster immunization was given subcutaneously with 100 μL of emulsion containing 50 μg bovine type II collagen and incomplete Freund's adjuvant. Mice were randomly divided into 5 groups (n=8 per group): Control group (normal mice, no treatment), CIA model group (CIA mice, administered with vehicle), Eleutheroside E 10 mg/kg group, Eleutheroside E 20 mg/kg group, Eleutheroside E 40 mg/kg group. Eleutheroside E was dissolved in normal saline containing 0.5% DMSO, and administered via intraperitoneal injection (0.1 mL/10 g body weight) once daily from day 21 to day 41. Arthritis score was evaluated weekly: each paw was scored on a scale of 0-4 (0: no swelling/redness; 1: mild swelling/redness of toes; 2: moderate swelling/redness of paw; 3: severe swelling/redness of paw; 4: complete loss of joint function), and the total score was the sum of four paws. At the end of the experiment, mice were sacrificed by cervical dislocation. Serum was collected by orbital blood sampling and centrifugation (3000×g for 15 minutes). Joint tissues (hind paws) were fixed in 4% paraformaldehyde for pathological sectioning, and spleens were weighed to calculate the spleen index (spleen weight/body weight × 1000) [1]
2. db/db mouse model protocol: Male db/db mice (8 weeks old) and age-matched C57BL/6J mice (control group) were used. Mice were randomly divided into 5 groups (n=8 per group): Control group (C57BL/6J mice, administered with vehicle), db/db model group (db/db mice, administered with vehicle), Eleutheroside E 5 mg/kg group, Eleutheroside E 10 mg/kg group, Eleutheroside E 20 mg/kg group. Eleutheroside E was dissolved in 0.5% carboxymethyl cellulose sodium solution, and administered via oral gavage (0.1 mL/10 g body weight) once daily for 4 weeks. Body weight was measured weekly. Fasting blood glucose (FBG) was measured weekly using a glucose meter (after 6 hours of fasting). At the end of the experiment, mice were fasted for 6 hours, and fasting insulin was detected by ELISA. HOMA-IR was calculated as (FBG × fasting insulin)/22.5. For glucose tolerance test (GTT), mice were fasted for 6 hours, then administered with glucose (2 g/kg body weight) via oral gavage. Blood glucose was measured at 0, 30, 60, 90, and 120 minutes after glucose administration. After GTT, mice were sacrificed, and adipose tissues (epididymal fat) and liver tissues were collected and stored at -80°C for subsequent Western blot analysis [2]

1. In the CIA mouse model: At the end of the experiment, the levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), blood urea nitrogen (BUN), and creatinine (Cr) in serum were detected using a biochemical kit. The results showed that there were no significant differences in the levels of ALT, AST, BUN, and Cr between the treatment group and the control group, indicating that Acanthopanax senticosin E (dose up to 40 mg/kg, intraperitoneal injection, for 21 consecutive days) had no significant hepatotoxicity or nephrotoxicity in mice [1]. 2. In the db/db mouse model: After administration of Acanthopanax senticosin E (dose up to 20 mg/kg, gavage) for 4 weeks, the levels of ALT, AST, BUN, and Cr in serum were detected. No significant changes were observed in these biochemical indicators compared with the control group, indicating that Acanthopanax senticosin E had no detectable toxicity in db/db mice [2].

[1]. Eleutheroside E Ameliorates Arthritis Severity in Collagen-Induced Arthritis Mice Model by Suppressing Inflammatory Cytokine Release. Inflammation, Vol. 37, No. 5, October 2014 (# 2014)

[2]. Eleutheroside E, An Active Component of Eleutherococcus senticosus, Ameliorates Insulin Resistance in Type 2 Diabetic db/db Mice. Evid Based Complement Alternat Med. 2013; 2013: 934183.

According to reports, quercetin is found in white mistletoe, plants of the genus Mallotus, and other organisms with relevant data.
See also: Acanthopanax senticosin E (note moved to) Acanthopanax senticosin (note moved to)

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1. Acanthopanax senticosin E is the main active ingredient isolated from the rhizome of Acanthopanax senticosus (Siberian ginseng), a traditional herb often used to enhance immunity and reduce inflammation [1][2]
2. The anti-arthritis mechanism of Acanthopanax senticosin E is mainly related to its ability to inhibit the excessive production of pro-inflammatory cytokines (IL-1β, TNF-α, IL-6) in the immune system, thereby reducing joint inflammation and tissue damage in CIA mice [1]
3. The mechanism by which Acanthopanax senticosin E improves insulin resistance in db/db mice involves activating the IRS-1/AKT/GLUT4 signaling pathway in adipose tissue, thereby enhancing insulin-mediated glucose uptake and metabolism [2]

C34H46O18

742.7183

742.268

39432-56-9

Eleutheroside D;79484-75-6

226371

Solid

1.5±0.1 g/cm3

935.7±65.0 °C at 760 mmHg

255 - 260ºC

519.7±34.3 °C

0.0±0.3 mmHg at 25°C

1.616

-3.91

8

18

12

52

1010

0

O1C([H])([H])[C@@]2([H])[C@]([H])(C3C([H])=C(C(=C(C=3[H])OC([H])([H])[H])O[C@]3([H])[C@]([H])([C@@]([H])([C@]([H])([C@]([H])(C([H])([H])O[H])O3)O[H])O[H])O[H])OC([H])([H])[H])OC([H])([H])[C@]2([H])[C@@]1([H])C1C([H])=C(C(=C(C=1[H])OC([H])([H])[H])O[C@@]1([H])[C@@]([H])([C@]([H])([C@@]([H])([C@@]([H])(C([H])([H])O[H])O1)O[H])O[H])O[H])OC([H])([H])[H]

FFDULTAFAQRACT-JSGUJALWSA-N

InChI=1S/C34H46O18/c1-43-17-5-13(6-18(44-2)31(17)51-33-27(41)25(39)23(37)21(9-35)49-33)29-15-11-48-30(16(15)12-47-29)14-7-19(45-3)32(20(8-14)46-4)52-34-28(42)26(40)24(38)22(10-36)50-34/h5-8,15-16,21-30,33-42H,9-12H2,1-4H3/t15-,16+,21-,22-,23-,24-,25+,26+,27-,28-,29+,30-,33+,34+/m1/s1

(2S,3R,4S,5S,6R)-2-[4-[(3S,3aR,6R,6aS)-6-[3,5-dimethoxy-4-[(2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxyphenyl]-1,3,3a,4,6,6a-hexahydrofuro[3,4-c]furan-3-yl]-2,6-dimethoxyphenoxy]-6-(hydroxymethyl)oxane-3,4,5-triol

Eleutheroside E

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)

DMSO: ~100 mg/mM (~134.6 mM)

Solubility in Formulation 1: ≥ 2.5 mg/mL (3.37 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 25.0 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: ≥ 2.5 mg/mL (3.37 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 25.0 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: ≥ 2.5 mg/mL (3.37 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 25.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.

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