Pioglitazone

Manufactured by Cayman Chemical

Sourced in United States

Pioglitazone is a chemical compound used in research and laboratory settings. It is a peroxisome proliferator-activated receptor gamma (PPAR-gamma) agonist. Pioglitazone can be used as a reference standard or for the development of analytical methods.

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Pioglitazone (Pio, (2 citations)

31 protocols using pioglitazone

Macrophage Polarization by PPARγ Modulation

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Bone marrow cells isolated from the femur and tibia of PPARγ-MKO or PPARγ-WT mice were matured in RPMI with 20% FBS, 1% Pen-Strep and 50 ng/mL recombinant mouse M-CSF for seven days, which results in a population that is >95% macrophages[12 (link)]. Following differentiation, bone marrow-derived macrophages (BMDM) were treated with the PPARγ agonist pioglitazone (10 μM), the PPARγ antagonist T0070907 (T007: 10 μM, Cayman), a combination of pioglitazone and T007, or vehicle control (DMSO) for 48 hours. In experiments where tumor homogenate was used to stimulate BMDM, whole left lungs were collected from tumor-bearing or naïve mice and snap frozen in liquid nitrogen. Tissues were homogenized in phosphate-buffered saline (PBS) using an overhead stirrer (Wheaton). A final concentration of 0.5 mg/mL protein each from tumor or control homogenate was added to BMDM for 48 hours. BMDM were then washed three times with HBSS and placed into fresh RPMI for 24 hours prior to collecting the culture media. TGF-β1 released into the media was measured by ELISA (R&D Systems). TGF-β1 levels were normalized to total BMDM RNA or protein concentration.

Sippel T.R., Johnson A.M., Li H.Y., Hanson D., Nguyen T.T., Bullock B.L., Poczobutt J.M., Kwak J.W., Kleczko E.K., Weiser-Evans M.C, & Nemenoff R.A. (2019). Activation of PPARγ in Myeloid Cells Promotes Progression of Epithelial Lung Tumors through TGF- β1. Molecular cancer research : MCR, 17(8), 1748-1758.

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Investigating Rosiglitazone, Pioglitazone, and GW9662 Effects

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Rosiglitazone, pioglitazone and GW9662 were purchased from Cayman Chemical (Ann Arbor, MI, United States). LPS and tunicamycin were purchased from Sigma-Aldrich (St. Louis, MO, United States). Rabbit anti-NGBR, β-actin, and GAPDH polyclonal antibodies were purchased from Abcam (Cambridge, MA, United States). Rabbit anti-IL-1β and IL-6 polyclonal antibodies were purchased from ABclonal (Wuhan, China). Rabbit anti-PPARγ and AKT polyclonal antibodies and mouse anti-TNF-α and phospho-AKT (p-AKT, Ser473) monoclonal antibodies were purchased from Proteintech Group (Chicago, IL, United States). Rabbit anti-CHOP, BIP, ATF6, phospho-IRE1α (p-IRE1α, Ser724) and phospho-PERK (p-PERK, Thr982) polyclonal antibodies were purchased from Affinity Biosciences (Cincinnati, OH, United States). Silencer siRNA Construction Kit was purchased from Life Technologies (New York, United States). Transfection reagent Lipofectamine RNAiMAX and Lipofectamine 2000 were purchased from Thermo Fisher Scientific (Waltham, MA, United States).

Ma J., Zeng P., Liu L., Zhu M., Zheng J., Wang C., Zhao X., Hu W., Yang X., Duan Y., Han J., Miao Q.R, & Chen Y. (2022). Peroxisome Proliferator-Activated Receptor-Gamma Reduces ER Stress and Inflammation via Targeting NGBR Expression. Frontiers in Pharmacology, 12, 817784.

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Pharmacological Pretreatments in Rat TGI

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In the experiments involving pharmacological pretreatments, the dosage for Mdivi-1 was tested as previously reported [8 (link), 30 (link)], the dosage of GW9662 and pioglitazone were based on our previous studies [26 (link), 31 (link)]. One group of rats were treated intraperitoneally with Drp1 inhibitor Mdivi-1 (2.4 mg/kg), which was purchased from Sigma-Aldrich Ltd (St. Louis, MO, USA), or the solvent dimethyl sulfoxide (DMSO) 30 min before TGI. The other group of rats were microinjected into bilateral CA1 subfields with pioglitazone (Cayman Chemical, Ann Arbor, MI, USA; 20 nmol), GW9662 (Cayman Chemical, 500 ng) or DMSO as the vehicle and volume control 30 min before TGI. The test agents were microinjected bilaterally in a volume of 100 nl on each side. Drug delivery into the hippocampal CA1 subfield was carried out as previously reported [18 (link), 19 (link)]. The animals receiving chloral hydrate anesthesia and surgical preparations without additional experimental manipulations served as sham-controls.

Chuang Y.C., Lin T.K., Yang D.I., Yang J.L., Liou C.W, & Chen S.D. (2016). Peroxisome proliferator-activated receptor-gamma dependent pathway reduces the phosphorylation of dynamin-related protein 1 and ameliorates hippocampal injury induced by global ischemia in rats. Journal of Biomedical Science, 23, 44.

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Neuroprotective Compound Evaluation in Microglial Cell Cultures

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Neurobasal-A medium, Dulbecco’s Modified Eagle’s Medium,
fetal bovine serum, B-27 supplement with and without antioxidant, fungizone,
gentamicin, and goat anti-rabbit IgG Alexa 55 were purchased from Invitrogen
(Thermo Fisher Scientific, Waltham, MA). Pioglitazone and minocycline
hydrochloride were purchased from Cayman Chemical (Ann Arbor, MI). Melatonin,
phenyl-α-tert-butyl nitrone (PBN) and
2',7'-dichlorodihydrofluorescein diacetate (DCF-DA) were
purchased from Sigma-Aldrich (St. Louis, MO). Rabbit polyclonal antibodies to
Iba1 and to paraoxonase-2 were purchased from Abcam (Cambridge, MA). Falcon
24-well cell culture inserts and PET membranes were purchased from Corning Life
Sciences (Tewksbury, MA). For lactate dehydrogenase assays, Promega’s
CytoTox 96 Non-Radioactive Cytotoxicity kit (Promega Corporation, Madison, WI)
was used. The Live/Dead Viability/Cytotoxicity Kit for mammalian cells was
purchased from Molecular Probes (Thermo Fisher Scientific, Waltham, MA). All
reagents for quantitative polymerase chain reaction (qPCR) were obtained from
Biorad Laboratories (Hercules, CA). The Quantikine ELISA Mouse IL-6 Immunoassay
kit was from R&D Systems, Inc. (Minneapolis, MN).

Roqué P.J., Dao K, & Costa L.G. (2016). Microglia mediate diesel exhaust particle-induced cerebellar neuronal toxicity through neuroinflammatory mechanisms. Neurotoxicology, 56, 204-214.

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Apoptosis Induction by PPARγ Antagonists

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Thiazolidinediones (rosiglitazone, pioglitazone, ciglitazone) and the PPARγ antagonist GW9662 were provided from Cayman Chemical (Ann Arbor, MI, USA). Soluble recombinant TRAIL, proteasome inhibitor MG132, caspase 9 specific inhibitor (Z-LEHD-FMK) and propidium iodide (PI) were purchased from Sigma (Saint Quentin Fallavier, France). Caspase 8 inhibitor (Z-IETD-FMK) was from Bachem (Heidelberg, Germany). Anti-DR4 and DR5 blocking antibodies were from Alexis Biochemicals (Lausanne, Switzerland).

Plissonnier M.L., Fauconnet S., Bittard H., Mougin C., Rommelaere J, & Lascombe I. (2017). Cell death and restoration of TRAIL-sensitivity by ciglitazone in resistant cervical cancer cells. Oncotarget, 8(64), 107744-107762.

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CYP26A1 and CYP26B1 Enzyme Assay

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CYP26A1 and CYP26B1 were generous gifts from Dr. Nina Isoherranen (University of Washington). Recombinant CYP2C8 Supersomes® and purified human cytochrome P450 reductase were obtained from Corning Life Sciences (Tewksbury, MA). Tazarotenic acid, MM11253, liarazole, EC23, AM80 and candesartan were purchased from Tocris Chemicals (Bristol, United Kingdom). Montelukast, pioglitazone, rosiglitazone and zafirlukast were from Cayman Chemical (Ann Arbor, MI). Talarazole was purchased from MedChem Express (Monmouth Junction, NJ). Rapid equilibrium dialysis (RED) device kits were obtained from ThermoFisher Scientific (Waltham, MA). All other chemicals were from Sigma-Aldrich (St. Louis, MO) and were of the highest grade available.

Foti R.S., Diaz P, & Douguet D. (2016). Comparison of the Ligand Binding Site of CYP2C8 with CYP26A1 and CYP26B1: A Structural Basis for the Identification of New Inhibitors of the Retinoic Acid Hydroxylases. Journal of enzyme inhibition and medicinal chemistry, 31(SUP2), 148-161.

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Telomerase Inhibition in Cancer Cells

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MCT (Must Bio-technology, Chengdu, China) was used to establish PH. BIBR1532 (APExBIO Techonologies, Houston, TX, USA) was used to impede the activity of telomerase. LY294002 (HY-MedChemExpress, Monmouth, NJ, USA). Pioglitazone (Cayman Chemical Company, Michigan, USA), was used to active PPAR-γ. Antibody against Akt, phospho-Akt (Ser473), c-Myc and GAPDH were purchased from Cell signaling Technology (Danvers, MA, USA). Polyclonal antibody against TERT were provided by Abcam (Cambridge, MA, USA).

Hussain T., Chai L., Wang Y., Zhang Q., Wang J., Shi W., Wang Q., Li M, & Xie X. (2023). Activation of PPAR-γ prevents TERT-mediated pulmonary vascular remodeling in MCT-induced pulmonary hypertension. Heliyon, 9(3), e14173.

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ROS Modulation by Pharmacological Agents

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Pioglitazone (5, 10, and 20 μM) (Cayman), GW9662 (10, 20 μM) (Cayman), a-lipoic acid (50, 200, and400 μM), and coenzyme Q10 (CoQ10) (0.5, 1, and 5 μM) were used for assessment of ROS generation. We optimized the concentration of each component individually in the production of ROS, by measuring fluorescence intensity using fluorescent microscopy, as well as flow cytometry.

Baharlooie M., Peymani M., Nasr-Esfahani M.H, & Ghaedi K. (2022). Pioglitazone Mediates Cardiac Progenitor Formation through Increasing ROS Levels. BioMed Research International, 2022, 1480345.

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3T3-L1 Cell Differentiation Assay

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DMEM, OptiMEM, penicillin-streptomycin, PBS and sodium pyruvate were obtained from Gibco by Life Technologies (Carlsbad, CA, United States), SYBR green MicroAmp Fast Optical 96-well Reaction Plate 0.1 ml and StepOnePlus thermocycler were obtained from Applied Biosystems (Foster City, CA, United States). FCS was purchased from Capricorn (Ebsdorfergrund, Germany). New born calf serum was purchased from Biochrom (Berlin, Germany). All primer pairs were purchased from Eurofins (Friedrichsdorf, Germany). Cell culture flasks (T75, Cell+, vented cap) and 6-well plates for sub-culturing 3T3-L1 cells were obtained from SARSTEDT (Nümbrecht, Germany). Rosiglitazone, pioglitazone, zafirlukast, montelukast, IBMX and (±)14(15)-EET-d₁₁ were purchased from Cayman Chemical (Ann Arbor, MI, United States). Insulin, dexamethasone and Oil Red O were obtained from Sigma Aldrich (St. Louis, Missouri, MO, United States). Tris, Triton-X-100, NP-40, NaCl, EDTA and SDS were purchased from AppliChem (Darmstadt, Germany).

Göbel T., Diehl O., Heering J., Merk D., Angioni C., Wittmann S.K., Buscato E.L., Kottke R., Weizel L., Schader T., Maier T.J., Geisslinger G., Schubert-Zsilavecz M., Steinhilber D., Proschak E, & Kahnt A.S. (2019). Zafirlukast Is a Dual Modulator of Human Soluble Epoxide Hydrolase and Peroxisome Proliferator-Activated Receptor γ. Frontiers in Pharmacology, 10, 263.

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Pioglitazone Modulates Sepsis Response

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For in vivo experiments, pioglitazone (20 mg/Kg, i.p) was administered 1, 4 and 18 hours before CLP surgery.
For in vitro experiments, pioglitazone, rosiglitazone, troglitazone, and GW9662 (10 μM each, Cayman Chemical) were incubated with 2×10⁶ cells for 24 h before immunoblotting or PCR assays. Anti-IL-10R (Biolegend, 20 μg/mL) was incubated for 0.5 h before pioglitazone and/or 100 ng/mL LPS (Sigma-Aldrich) stimulation.

Ferreira A.E., Sisti F., Sônego F., Wang S., Filgueiras L., Brandt S., Serezani A.P., Du H., Cunha F.Q., Alves-Filho J.C, & Serezani C.H. (2014). PPAR-γ/IL-10 axis inhibits MyD88 expression and ameliorates murine polymicrobial sepsis. Journal of immunology (Baltimore, Md. : 1950), 192(5), 2357-2365.

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