After 18α-glycyrrhetinic acid (18a-GA) therapy for 48 hours and 72 hours, the number of LX-2 cells was dramatically reduced by 14.8% and 31.2% correspondingly (P<0.05). Compared with the control group, after 48 and 72 hours of 18α-glycyrrhetinic acid administration, the proportion of LX-2 cells in the G0/G1 phase considerably increased, while the percentage of cells in the S phase decreased. 18α-Glycyrrhetinic acid increased cell apoptosis to 6.8% after 48 hours compared to the control group (2.5%), and at 72 hours, the percentage of apoptotic cells in LX-2 cells in the control and treatment groups were respectively 3.1% and 15.6% (P<0.01). In addition, 18α-glycyrrhetinic acid stimulates the expression of PPAR-γ and modifies several cell cycle and apoptosis-related proteins. 18α-Glycyrrhetinic acid also suppresses NF-κB DNA binding activity [1].

After 18α-glycyrrhetinic acid (18a-GA) therapy for 48 hours and 72 hours, the number of LX-2 cells was dramatically reduced by 14.8% and 31.2% correspondingly (P<0.05). Compared with the control group, after 48 and 72 hours of 18α-glycyrrhetinic acid administration, the proportion of LX-2 cells in the G0/G1 phase considerably increased, while the percentage of cells in the S phase decreased. 18α-Glycyrrhetinic acid increased cell apoptosis to 6.8% after 48 hours compared to the control group (2.5%), and at 72 hours, the percentage of apoptotic cells in LX-2 cells in the control and treatment groups were respectively 3.1% and 15.6% (P<0.01). In addition, 18α-glycyrrhetinic acid stimulates the expression of PPAR-γ and modifies several cell cycle and apoptosis-related proteins. 18α-Glycyrrhetinic acid also suppresses NF-κB DNA binding activity [1].

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18α-Glycyrrhetinic acid (8 µM) treatment for 48 h and 72 h significantly reduced the number of human hepatic stellate cell line LX-2 by 14.8% and 31.2%, respectively, compared to the control group. A similar inhibitory effect was observed on rat cirrhotic fat-storing cells (CFSC). [1]
Flow cytometric analysis showed that 18α-Glycyrrhetinic acid (8 µM) increased the percentage of LX-2 and CFSC cells in the G0/G1 phase and decreased the percentage in the S phase after 48 h and 72 h of treatment. [1]
18α-Glycyrrhetinic acid (8 µM) induced apoptosis in LX-2 cells, with apoptotic rates of 6.8% at 48 h and 15.6% at 72 h, compared to 2.5% and 3.1% in the control group, respectively. Similar pro-apoptotic effects were observed in CFSC cells. [1]
Real-time PCR and Western blot analysis demonstrated that 18α-Glycyrrhetinic acid (8 µM) treatment increased the mRNA and protein levels of PPAR-γ in a time-dependent manner. [1]
Western blot analysis revealed that 18α-Glycyrrhetinic acid (8 µM) decreased the protein level of cyclin D1 and increased the level of p27 in LX-2 cells. It also downregulated the anti-apoptotic proteins Bcl-xl and Bfl/A1, while upregulating the pro-apoptotic protein Bax. [1]
A transcription factor activity assay showed that 18α-Glycyrrhetinic acid (8 µM) significantly inhibited NF-κB DNA-binding activity in LX-2 cells by 8% at 24 h and 17% at 48 h. [1]
All the above effects of 18α-Glycyrrhetinic acid on cell proliferation, apoptosis, cell cycle, protein expression, and NF-κB activity were abolished by pre-treatment with the selective PPAR-γ antagonist GW9662 (0.5 µM). [1]

18α-glycyrrhetinic acid (18α-GA) treatment can greatly increase the longevity of nematode strains; 20 μg/mL is the most effective quantity. The outcomes demonstrated a considerable delay in the onset of paralysis symptoms following treatment with 18α-glycyrrhetinic acid. Treatment with 18α-glycyrrhetinic acid can also considerably lower Aβ deposition [2].

Cell Viability and Proliferation Assay: Human LX-2 and rat CFSC cells were seeded in 96-well plates. After adherence, cells were treated with 18α-Glycyrrhetinic acid at concentrations ranging from 0 to 32 µM for 48 hours. Cell viability was assessed using a Cell Counting Kit-8 (CCK-8). Absorbance was measured at 450 nm. A concentration of 8 µM was selected for subsequent experiments as cell viability remained above 80%. [1]
Cell Growth Determination: Cells were seeded in 24-well plates, starved for 24 hours, and then stimulated with medium containing 10% fetal bovine serum with or without 8 µM 18α-Glycyrrhetinic acid. Cell numbers were counted at 24, 48, and 72 hours post-treatment using a computer-equipped cell counter. [1]
Cell Cycle Analysis by Flow Cytometry: Cells were seeded in 6-well plates, starved, and treated with 8 µM 18α-Glycyrrhetinic acid for 24, 48, and 72 hours. Cells were then harvested, and their DNA content was analyzed by flow cytometry to determine the distribution across different cell cycle phases (G0/G1, S, G2/M). [1]
Apoptosis Analysis by Flow Cytometry: After treatment with 8 µM 18α-Glycyrrhetinic acid for specified times, cells were stained using an Annexin V-based staining kit according to the manufacturer's instructions. The percentage of apoptotic cells was quantified by flow cytometry. [1]
Gene Expression Analysis (Real-time PCR): Total RNA was extracted from treated cells. Reverse transcription was performed. The mRNA levels of PPAR-γ were quantified by real-time PCR using SYBR Green, with GAPDH serving as the internal control. The fold change was calculated. [1]
Protein Expression Analysis (Western Blot): Total protein was extracted from cultured cells. Proteins were separated by SDS-PAGE, transferred to membranes, and probed with specific primary antibodies against various targets (e.g., cyclins, p21, p27, PPAR-γ, Bcl-2 family proteins). GAPDH was used as a loading control. Protein bands were visualized using enhanced chemiluminescence. [1]
NF-κB DNA-Binding Activity Assay: Nuclear proteins were extracted from treated cells. Equal amounts of nuclear protein were added to wells coated with an oligonucleotide containing the NF-κB consensus binding site. The binding of active NF-κB (p65 subunit) was detected using a specific primary antibody, followed by a horseradish peroxidase-conjugated secondary antibody, and quantified colorimetrically at 450 nm. [1]

Cytotoxicity: In cell viability assays using the CCK-8 assay, both LX-2 and CFSC cell lines showed viability above 80% after treatment with 16 µM 18α-glycyrrhetinic acid for 48 hours. Higher concentrations may show higher toxicity. [1]

[1]. 18α-glycyrrhetinic acid extracted from Glycyrrhiza radix inhibits proliferation and promotes apoptosis of the hepatic stellate cell line. J Dig Dis. 2013 Jun;14(6):328-36.

[2]. 18α-Glycyrrhetinic Acid Proteasome Activator Decelerates Aging and Alzheimer's Disease Progression in Caenorhabditis elegans and Neuronal Cultures. Antioxid Redox Signal. 2016 Dec 1;25(16):855-869.

Glycyrrhetinic acid is a pentacyclic triterpenoid compound formed by the substitution of oleanolic-12-ene at positions 3, 11, and 30 with hydroxyl, carbonyl, and carboxyl groups, respectively. It possesses immunomodulatory effects and is also a plant metabolite. It is a pentacyclic triterpenoid compound, a cyclic terpene ketone, and a hydroxy monocarboxylic acid. It is the conjugate acid of glycyrrhetinic acid. It is derived from the hydride of oleanolic acid. Enoxetine (glycyrrhizic acid) has been used in basic scientific research on epigenetic mineralocorticoid hyperparathyroidism (AME). Enoxetine has been reported to exist in plants of the genus Glycyrrhiza (such as Glycyrrhiza glabra) and other organisms with relevant data. Enoxetine is a pentacyclic triterpenoid aglycone metabolite of glycyrrhizic acid, a product of the plant Glycyrrhiza glabra, which has potential expectorant and prokinetic effects. After administration, enoxetine can inhibit prostaglandin metabolism by inhibiting 15-hydroxyprostaglandin dehydrogenase [NAD(+)] and prostaglandin reductase 2. Therefore, this drug can enhance the activity of prostaglandins E2 and F2α, thereby inhibiting gastric acid secretion, while stimulating pancreatic secretion and the secretion of intestinal and respiratory mucus, ultimately leading to enhanced intestinal motility and antitussive effects. In addition, this drug can also inhibit 11β-hydroxysteroid dehydrogenase and other enzymes in the kidneys involved in the conversion of cortisol to cortisone.
Glycyrrhizic acid is an oleanolic acid derived from licorice, possessing certain anti-allergic, antibacterial, and antiviral properties. It can be used topically to treat allergic or infectious skin inflammations, or orally to exert its aldosterone effect in electrolyte regulation.
See also: glycyrrhizic acid (its active ingredient); ammonium glycyrrhizate (active ingredient); licorice root extract (partial components).
18α-Glycyrrhetinic acid is the epimer of 18β-glycyrrhetinic acid at the 18-H position. Compared to 18β-glycyrrhetinic acid, 18α-glycyrrhetinic acid has stronger targeting to the liver, stronger hepatocyte protection, and lower toxicity. [1]
Studies have shown that the antiproliferative and pro-apoptotic effects of 18α-glycyrrhetinic acid on hepatic stellate cells are mediated at least in part by upregulating PPAR-γ expression and subsequently inhibiting NF-κB activity. This highlights its potential as a candidate drug for antifibrotic therapy of liver fibrosis. [1]

C30H46O4

470.68384

470.339

1449-05-4

18β-Glycyrrhetinic acid;471-53-4

10114

White to off-white solid powder

1.1±0.1 g/cm3

588.3±50.0 °C at 760 mmHg

331-333°C

323.7±26.6 °C

0.0±3.7 mmHg at 25°C

1.563

6.57

2

4

1

34

965

9

C[C@]12CC[C@](C[C@H]1C3=CC(=O)[C@@H]4[C@]5(CC[C@@H](C([C@@H]5CC[C@]4([C@@]3(CC2)C)C)(C)C)O)C)(C)C(=O)O

MPDGHEJMBKOTSU-YKLVYJNSSA-N

InChI=1S/C30H46O4/c1-25(2)21-8-11-30(7)23(28(21,5)10-9-22(25)32)20(31)16-18-19-17-27(4,24(33)34)13-12-26(19,3)14-15-29(18,30)6/h16,19,21-23,32H,8-15,17H2,1-7H3,(H,33,34)/t19-,21-,22-,23+,26+,27-,28-,29+,30+/m0/s1

(2S,4aS,6aR,6aS,6bR,8aR,10S,12aS,14bR)-10-hydroxy-2,4a,6a,6b,9,9,12a-heptamethyl-13-oxo-3,4,5,6,6a,7,8,8a,10,11,12,14b-dodecahydro-1H-picene-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

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

DMSO : ~11.36 mg/mL (~24.14 mM)

Solubility in Formulation 1: 1.14 mg/mL (2.42 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 11.4 mg/mL clear DMSO stock solution to 400 μL of PEG300 and mix evenly; then add 50 μL of Tween-80 to the above solution and mix evenly; then add 450 μL of 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: ≥ 1.14 mg/mL (2.42 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 11.4 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.)