All experiments involving mice were approved by the Animal Experiment Administration Committee of the Fourth Military Medical University. 5 × 106 patient-derived GBM cells suspended in 100 μL PBS were inoculated subcutaneously into the right forelimb interior root of BABL/c-A nude mice (female) at 4 weeks of age. About 7–8 days after cell implantation, the mice bearing tumor around 50 mm3 were randomly divided into a control group, glycyrrhizin group, TMZ group or TMZ + glycyrrhizin group. Mice in the control group received equivalent drug vehicle (dimethyl sulfoxide, DMSO), mice in glycyrrhizin group received 10 mg/kg glycyrrhizin (SelleckChem, Houston TX, USA) five times per week for 2 weeks (intraperitioneal injection, i.p.), mice in TMZ group received 5 mg/kg TMZ five times per week for 3 weeks (i.p.), and mice in TMZ + glycyrrhizin group received 10 mg/kg glycyrrhizin five times per week for 2 weeks and also 5 mg/kg TMZ five times per week for 3 weeks (i.p.). Tumor volume was measured every 3 days with a caliper and calculated using the formula tumor volume (mm3) = (length × width2) /2. About 24 days after the first treatment, all mice were euthanized and the tumor were carefully removed, weighed.
Glycyrrhizin
Manufactured by Selleck Chemicals
Sourced in United States
Glycyrrhizin is a naturally occurring compound extracted from the roots of the licorice plant (Glycyrrhiza glabra). It is a complex organic compound with a chemical formula of C42H62O16. Glycyrrhizin exhibits various biological properties and is commonly used in pharmaceutical and research applications.
Automatically generated - may contain errors
6 protocols using glycyrrhizin
1
Glioblastoma Xenograft Tumor Model
2
Glycyrrhizin Protects Contrast-Induced Toxicity
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The NRK52E cell line (immortalized normal rat renal proximal tubular cells) were cultured in DMEM with 5% FBS and 100U/ml penicillin–streptomycin at 37 °C in a humidified incubator with 5% CO2 for growth. When the cell confluence was 85%, the cells were exposed to 200 mg I/ml contrast media (Pamiray 370, Dongkook Lifescience, Seoul, South Korea) for 2 h. The glycyrrhizin pretreatment group was pretreated with 150 uM of glycyrrhizin (Selleckchem, Huston, TX) 90 min before contrast media exposure.
3
Intrauterine Injection of HMGB1 Inhibitor
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Intrauterine injection of HMGB1 inhibitor was performed as previously described (13 (link)). At 09:00 on the day 4 of pregnancy, two uterine horns of each mouse were slowly injected with Glycyrrhizin (3 μl per uterine horn, 100 μM in saline, NSC 167409, Selleck), a specific HMGB1 inhibitor. In control group, two uterine horns of each mouse were injected with an equal volume of normal saline (3 μl per uterine horn).
4
Glycyrrhizin Administration for Wound Healing
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Before use, glycyrrhizin (Selleck Chemicals LLC, USA) was dissolved in 50 mM NaOH at 37 °C, and the pH was adjusted to 7.4 using 0.5 M Tris–HCl buffer (pH 6.8). Immediately after the wound was sutured, glycyrrhizin (1 mg/kg) was injected intraperitoneally and then administered intraperitoneally daily for one week. The dose was determined as described previously [26 (link)].
5
Biochemical Assay Protocol for Inflammatory and Metabolic Markers
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Cu was purchased from Sigma Aldrich (Saint Louis, MO). Glucose was obtained from Sigma Aldrich (St. Louis, MO). Glycyrrhizin was purchased from Selleck (USA). Enzyme-linked immunosorbent assay kits of Interleukin (IL)-6, IL-1β, and tumor necrosis factor (TNF)-α were purchased from Elabscience (Wuhan, China). The commercial kits of total triglycerides (TGs), total cholesterol (TC), creatine kinase (CK), and lactate dehydrogenase (LDH) were purchased from Jiancheng Bioengineering Institute (Nanjing, China). Primary antibodies were purchased from Cell Signaling Technology.
6
Culturing Mouse Cancer and Myoblast Cells
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Mouse CT26 colon cancer cells (CT26 cells) and mouse myoblasts (C2C12 cells) were purchased from Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences. CT26 cells and C2C12 were cultured in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% fetal bovine serum (FBS), 1% penicillin G-streptomycin (Gibco, MA, United States) in 37°C incubators with 5% CO2. When C2C12 myoblasts reached 90%, the proliferation medium was discarded and replaced with DMEM medium containing 2% horse serum (Gibco, MA, United States). The differentiation medium was replaced every day. Glycyrrhizin was obtained from SelleckChem (Huston TX, United States) and dissolved in dimethyl sulfoxide (DMSO). TAK-242, BAY 11-7082, PD169316 were purchased from MedChem Express (NJ, United States).
When the cell density of CT26 reaches 80%, discard the complete medium and wash with sterile PBS gently for at least 3 times. Then continue to incubate for 48 h with serum-free DMEM. The CT26 cell supernatant was collected and centrifuged at 1,200 rpm for 10 min to remove the cell debris. Filter the centrifuged CT26 supernatant through a 0.2 μm sterile syringe filter and the medium was used as a cachectic factor. CT26 conditioned medium (CCM) were prepared by CT26 supernatant and DMEM at a ratio of 1:1 with 2% horse serum.
When the cell density of CT26 reaches 80%, discard the complete medium and wash with sterile PBS gently for at least 3 times. Then continue to incubate for 48 h with serum-free DMEM. The CT26 cell supernatant was collected and centrifuged at 1,200 rpm for 10 min to remove the cell debris. Filter the centrifuged CT26 supernatant through a 0.2 μm sterile syringe filter and the medium was used as a cachectic factor. CT26 conditioned medium (CCM) were prepared by CT26 supernatant and DMEM at a ratio of 1:1 with 2% horse serum.
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