Metformin

Manufactured by Beyotime

Sourced in China

Metformin is a pharmaceutical compound used as a laboratory reagent. It is a biguanide drug primarily employed in the treatment of type 2 diabetes. Metformin functions by reducing hepatic glucose production and improving insulin sensitivity.

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Lab products found in correlation

Bend 3, by American Type Culture Collection (1 mentions) Dexamethasone (dex), by Merck Group (1 mentions) Trastuzumab, by Roche (1 mentions) Cd206, by Proteintech (1 mentions) Recombinant murine ifn γ, by Beyotime (1 mentions) Apc anti mouse cd206 antibody, by Elabscience (1 mentions) Ampkα d63g4 rabbit mab, by Cell Signaling Technology (1 mentions) Phospho ampkα thr172 40h9 rabbit mab, by Cell Signaling Technology (1 mentions) β actin, by Cell Signaling Technology (1 mentions) Lipopolysaccharide, by Beyotime (1 mentions) Microplate analyzer, by Thermo Fisher Scientific (1 mentions) Western blotting imaging system, by Bio-Rad (1 mentions) Fetal bovine serum (fbs), by Merck Group (1 mentions) Glyceraldehyde 3 phosphate dehydrogenase gapdh, by Affinity Biosciences (1 mentions) Cleaved caspase 3, by Affinity Biosciences (1 mentions) Annexin 5 fluorescein isothiocyanate fitc propidium iodide apoptosis assay kit, by BD (1 mentions) Cell counting kit 8 cck 8, by Beyotime (1 mentions) Dichlorofluorescein diacetate, by Thermo Fisher Scientific (1 mentions) 8 br camp, by Merck Group (1 mentions) Scintillation cocktail, by PerkinElmer (1 mentions)

11 protocols using metformin

Metformin Modulates LPS-Induced Endothelial Inflammation

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Mouse brain microvascular endothelial cells bEND3 (American Type Culture Collection, Rockville, MD, USA) were grown as a monolayer in DMEM with 15% fetal bovine serum (FBS), 100 U mL⁻¹ penicillin, and 100 μg mL⁻¹ streptomycin at 37°C in a humidified incubators with 5% CO₂ and 95% room air. Cells were subcultured into 35‐mm dishes and allowed to grow to confluence before incubating with metformin.
The confluent endothelial cells were first treated with metformin (Beyotime, Shanghai, China) at a dose of 0, 0.1, 0.5, and 1 mm for 24 h to define the optimal dose in activating AMPK. Then, the cells were treated with metformin at the optimal dose for 8 h before adding 1 μg mL⁻¹ LPS (Zhao et al., 2014), and 16 h later, Western blot was used to detect gp91^phox protein levels.

Wang X., Xue G., Liu W., Shu H., Wang M., Sun Y., Liu X., Sun Y.E., Liu C., Liu J., Liu W, & Jin X. (2017). Melatonin alleviates lipopolysaccharide‐compromised integrity of blood–brain barrier through activating AMP‐activated protein kinase in old mice. Aging Cell, 16(2), 414-421.

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Molecular Compounds for Cancer Treatment

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The 2DG (Cat# HY-13966), rosiglitazone (Cat# HY-17386), and pioglitazone (Cat# HY-13956) were obtained from MCE (MedChemExpress). Trastuzumab was obtained from Roche Pharma (Schweiz) Ltd., metformin (Cat# S1741) from Beyotime, and dexamethasone (Cat# D4902) from Sigma-Aldrich.

Wang J., Huang Q., Hu X., Zhang S., Jiang Y., Yao G., Hu K., Xu X., Liang B., Wu Q., Ma Z., Wang Y., Wang C., Wu Z., Rong X., Liao W, & Shi M. (2022). Disrupting Circadian Rhythm via the PER1–HK2 Axis Reverses Trastuzumab Resistance in Gastric Cancer. Cancer Research, 82(8), 1503-1517.

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Macrophage Polarization Modulation Protocol

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Tetraethylorthosilicate (TEOS), Na₂CO₃, ethanol, methanol, ammoniumhydroxide (NH₃·H₂O), hexadecyl-trimethyl-ammoniumbromide (CTAB) and potassium permanganate (KMnO₄) were purchased from SinopharmChemReagent Co., Ltd. (China). Metformin, lipopoly-saccharide (LPS) and Recombinant Murine IFN-γ were purchased from Beyotime Biotechnology Co., Ltd. (Shanghai, China). Murine IL-4 was provided by PeproTech Biotechnology Co., Ltd. (Suzhou, China). Polyclonal antibodies CD47, CD80 and CD206 were obtained from Proteintech Group, Inc. (Wuhan, China). Coumarin-6 was ordered from Aladdin Biochemical Technology Co., Ltd. (Shanghai, China). Mouse Tumor Necrosis Factor Alpha and Mouse IL-10 ELISA kit was provided by ABclonal Biotechnology Co., Ltd. (Wuhan, China). ELISA kits of Mouse iNOS and Mouse Arg-1 were obtained from Jonln Biotechnology Co., Ltd. (Shanghai, China). FITC-Anti-Mouse CD80 Antibody, FITC-Anti-Mouse CD206 and APC-Anti-Mouse CD206 Antibody were provided by Elabscience Biotechnology Co., Ltd. (Wuhan, China). β-Actin, AMPKα (D63G4) Rabbit mAb and Phospho-AMPKα (Thr172) (40H9) Rabbit mAb were ordered from Cell Signaling Technology, Inc. (MA, USA). DSPE-PEG-M2pep was purchased from SunLipo NanoTech Co., Ltd. (Shanghai, China).

Chong L., Jiang Y.W., Wang D., Chang P., Xu K, & Li J. (2023). Targeting and repolarizing M2-like tumor-associated macrophage-mediated MR imaging and tumor immunotherapy by biomimetic nanoparticles. Journal of Nanobiotechnology, 21, 401.

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Metformin Modulates PI3K/Akt/mTOR in Bladder Cancer

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Human bladder cancer cell lines T24 and 5637 were provided by the cell bank of the Chinese Academy of Sciences in Shanghai. The present study used metformin (Shanghai Beyotime Biotechnology Co., Ltd., Shanghai, China), Roswell Park Memorial Institute (RPMI)-1640 medium (Cellmax Company, Groningen, Netherlands), fetal bovine serum (FBS) (Sigma, St. Louis, MO, USA), phosphate-buffered saline (PBS) (Beijing Solarbio Company, Beijing, China), Cell counting Kit-8 (CCK-8) (Shanghai Beyotime Biotechnology Co., Ltd., Shanghai, China), and an Annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) Apoptosis Assay Kit (BD Biosciences, San Jose, CA, USA). Antibodies recognizing PI3K/Akt/mTOR signaling pathway associated proteins (phosphatidylinositol-4,5-bisphosphate 3-kinase (PI3K), phosphorylated (p)-PI3K, protein kinase B (Akt), p-Akt, mechanistic target of rapamycin (mTOR), p-mTOR), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and Apoptosis-related protein (cleaved-caspase 3 and cleaved-poly(ADP-ribose) polymerase (PARP)) were all purchased from Affinity Biosciences (Cincinnati, OH, USA). We also used a microplate analyzer (Thermo Scientific, Waltham, MA, USA), a flow cytometer (Bio-Rad, Hercules, CA, USA), and a western blotting imaging system (Bio-Rad).

Shen Z., Xue D., Wang K., Zhang F., Shi J., Jia B., Yang D., Zhang Q., Zhang S., Jiang H., Luo D., Li X., Zhong Q., Zhang J., Peng Z., Han Y., Sima C., He X, & Hao L. (2022). Metformin exerts an antitumor effect by inhibiting bladder cancer cell migration and growth, and promoting apoptosis through the PI3K/AKT/mTOR pathway. BMC Urology, 22, 79.

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Metformin Pretreatment in DSS-Induced Colitis

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Metformin (Beyotime Biotechnology) was reconstituted in normal saline. Mice were administered Metformin by daily i.p. injection at 250 mg/kg body weight consecutively for 7 d before being subjected to DSS-induced acute colitis.

Liu X., Lu J., Liu Z., Zhao J., Sun H., Wu N., Liu H., Liu W., Hu Z., Meng G., Shen L., Miller A.W., Su B., Li X, & Kang Z. (2018). Intestinal Epithelial Cell–Derived LKB1 Suppresses Colitogenic Microbiota. Journal of immunology (Baltimore, Md. : 1950), 200(5), 1889-1900.

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Cholesterol Metabolism Regulation Assay

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A-769662 was provided by Santa Cruz. AICAR and 8-Br-cAMP were purchased from Sigma and metformin from Beyotime Institute of Biotechnology. IMM-H007 was provided by the Institute of Material Medica, Chinese Academy of Medical Sciences and Peking Union Medical College (99.86% purity by HPLC). Acetylated (ac)LDL was obtained from Peking Union-Biology Co. Ltd. The [1,2-³H(N)]cholesterol and scintillation cocktails were purchased from PerkinElmer Life Sciences. The 1-palmitoyl-2-arachidonyl-sn-glycero-3-phosphorylcholine (PAPC) and hydroperoxyoctadecadienoic acid (HPODE) were purchased from Avanti Polar Lipids; dichlorofluorescein diacetate was purchased from Molecular Probes; and 1-palmitoyl-2-(5,6-epoxyisoprostaneE (2))-sn-glycero-3-phosphocholine was prepared from PAPC as described previously (19 (link), 20 (link)).

Ma A., Wang J., Yang L., An Y, & Zhu H. (2017). AMPK activation enhances the anti-atherogenic effects of high density lipoproteins in apoE−/− mice. Journal of Lipid Research, 58(8), 1536-1547.

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AMPK Pathway Modulation Impacts Fibrosis

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Metformin and 5-amino-4-imidazolecarboxamide riboside-1-b-D-ribofuranoside (AICAR) were bought from Beyotime Institute of Biotechnology (China). The antibodies used in this study include rabbit polyclonal anti-FN (H-300), rabbit polyclonal anti-SIRT1 (H-300), rabbit polyclonal anti-Egr1 (C-19), rabbit polyclonal anti-CTGF (H-55), and rabbit polyclonal anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (FL-335) from Santa Cruz Biotechnology (Santa Cruz, CA, USA) and rabbit monoclonal anti-AMPKα (4188S) and rabbit monoclonal anti-phospho-AMPKα (Thr172) (4811S) from Cell Signaling Technology (USA).

Wu C., Qin N., Ren H., Yang M., Liu S, & Wang Q. (2018). Metformin Regulating miR-34a Pathway to Inhibit Egr1 in Rat Mesangial Cells Cultured with High Glucose. International Journal of Endocrinology, 2018, 6462793.

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Silica-Induced Lung Injury and Metformin Therapy

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A total of 24 mice were randomly divided into four groups (n = 6 each group, using Excel 2010 software random number formula): a saline group, a silica group, a 100 mg/kg of metformin plus silica group, and a 300 mg/kg of metformin plus silica group. All the mice in the silica groups were received a single intratracheal instillation with 50 mg/kg of silica particles (size distribution: 99% between 0.5 and 10 μm, 80% between 1 and 5 μm, average particle diameter 1.7 μm, obtained from Sigma-Aldrich, St. Louis, MO, USA) dissolved in 0.05 ml sterile saline. After silica treatment for 28 days, the mice of metformin treatment groups were given a corresponding dose of metformin (started from the 28th day, Beyotime Institute of Biotechnology, Shanghai, China) via intragastrical administration daily for 2 weeks. Then the mice were sacrificed after installation, and the lung tissues were isolated and stored at – 80 °C for further analysis.

Cheng D., Xu Q., Wang Y., Li G., Sun W., Ma D., Zhou S., Liu Y., Han L, & Ni C. (2021). Metformin attenuates silica-induced pulmonary fibrosis via AMPK signaling. Journal of Translational Medicine, 19, 349.

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Investigating Metformin's Modulation of Cell Responses to Silica Nanoparticles

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The lung epithelial cells (A549 and HBE), lung fibroblast (MRC-5), and the human monocytic cell (THP-1) were commercially obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA). A549 and THP-1 cells were maintained in RPMI Medium 1640 basic (1640, Life Technologies/Gibco, Grand Island, NY, USA), the HBE cells were maintained in Dulbecco’s modified Eagle’s medium (D MEM, 1640, Life Technologies/Gibco, Grand Island, NY, USA), and the MRC-5 cells were maintained in Minimum Essential Medium (MEM, Life Technologies/Gibco, Grand Island, NY, USA). All of the culture media were containing 10% fetal bovine serum (BISH1475, Biological Industries) and antibiotics (penicillin and streptomycin, Life Technologies/Gibco, Gaithersburg, MD). Cells were cultured at 37 °C in a 5% CO₂ atmosphere.
PMA (Sigma–Aldrich) was used to treat THP-1 cells into macrophages. For all the experiments analysis, epithelial cells and THP-1 were treated with 200 or 150 μg/ml Silicon dioxide (SiO₂) (Sigma-Aldrich, St. Louis, MO, USA) together with various concentrations (2, 5, 10 mM) of metformin (Beyotime Institute of Biotechnology, Shanghai, China); MRC-5 cells were treated with 5 ng/ml TGF-β1 (Sigma-Aldrich) together with metformin.

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Isolation and Cultivation of Human Adipocytes

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Human adipocytes were acquired from fresh omentum of ovarian cancer patients undergoing surgery. Fresh omentum was minced with scissors, digested by gentleMACS (Miltenyi) [45 (link)] to obtain single cells. After centrifuge with condition of 100rpm, the suspension separated into 4 layers, which are cell pallets, conditional medium, adipocytes and free fat oil. And then we carefully isolated adipocytes, counted and cultured in six-well plates with DF12 (Gibco) supplemented with 2% FBS (Gibco) for 1×10⁶ adipocytes. 48 hours later, the supernatant was harvested as conditioned medium of human primary cultured adipocytes (hCM). hCM were filtered through 200-mesh sieves to remove cell debris before use.
3T3-L1 cells were chemically induced by DMEM with 10% FBS (Gibco), 0.5mM IBMX (TOPSCIENCE, T1713), 1μM Dexamethasone (TOPSCIENCE, T1076), 5μg/ml insulin (Bovine, Sigma I-5500) for 2 days and DMEM with 10% FBS (Gibco), 10μg/ml insulin (Bovine, Sigma I-5500) for 2 days to differentiate into adipocyte-like cells. Then the supernatant of induced adipocyte-like 3T3-L1 cells was collected at 24 hours as mouse conditioned medium (mCM). After pretreatment with metformin (1 mM, Beyotime, catalog number s1741–1g) for 12 hours, the induced adipocyte-like 3T3-L1 cells were cultured with complete medium for 24 hours, and the supernatant collected as CM (MET).

Sun C., Li X., Guo E., Li N., Zhou B., Lu H., Huang J., Xia M., Shan W., Wang B., Li K., Weng D., Xu X., Gao Q., Wang S., Hu J., Lu Y., Mills G.B, & Chen G. (2019). MCP-1/CCR-2 axis in adipocytes and cancer cell respectively facilitates ovarian cancer peritoneal metastasis. Oncogene, 39(8), 1681-1695.

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