All the included patients received dietary intervention and exercise therapy, other drugs affecting plasma glucose of the patients were avoided during the experiment. Medication: i) Group A (rosiglitazone group), taking rosiglitazone (4 mg; GlaxoSmithKline, Middlesex, UK), once a day; ii) group B (metformin group), taking metformin (0.15 g; Beijing Zhonghui Pharmaceutical Co., Ltd., Beijing, China) 3 times a day, the medicines were taken during or after meal to reduce gastrointestinal reactions; and iii) group C (rosiglitazone + metformin group), taking 4 mg rosiglitazone once a day and 0.15 g metformin 2 times a day. Medicine in the three groups were all oral administered lasting for 24 weeks.
Rosiglitazone
Manufactured by GlaxoSmithKline
Sourced in United Kingdom
Rosiglitazone is a pharmaceutical compound used as a laboratory reagent. It is a peroxisome proliferator-activated receptor gamma (PPAR-gamma) agonist. Rosiglitazone can be used in research studies to investigate the effects of PPAR-gamma activation on various cellular processes and biological functions.
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Lab products found in correlation
Dexamethasone (dex), by Merck Group (3 mentions)
3 isobutyl 1 methylxanthine, by Merck Group (2 mentions)
Insulin, by Merck Group (2 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Dorsomorphin, by Merck Group (1 mentions)
Pioglitazone, by Merck Group (1 mentions)
Penicillin streptomycin p s, by Thermo Fisher Scientific (1 mentions)
Propylene glycol, by Merck Group (1 mentions)
Phospho acc, by Cell Signaling Technology (1 mentions)
Pparγ, by Cell Signaling Technology (1 mentions)
C ebpα, by Cell Signaling Technology (1 mentions)
C ebpβ, by Cell Signaling Technology (1 mentions)
Phospho ampk, by Cell Signaling Technology (1 mentions)
Phospho akt, by Cell Signaling Technology (1 mentions)
Trypsin edta, by Thermo Fisher Scientific (1 mentions)
Gapdh, by Cell Signaling Technology (1 mentions)
Oil red o, by Merck Group (1 mentions)
Ethanol, by Merck Group (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
D glucose, by Merck Group (1 mentions)
9 protocols using rosiglitazone
1
Effects of Rosiglitazone and Metformin on Glycemic Control
2
Adipocyte Differentiation Assay
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d -glucose, 3-isobutyl-1-methylxanthine, dexamethasone, insulin, ethanol (EtOH), Oil-Red O, propylene glycol, dimethyl sulfoxide (DMSO), pioglitazone, metformin, dorsomorphin (an AMPK inhibitor, AI), and all other chemicals were of analytical grade and obtained from Sigma-Aldrich Inc. (St. Louis, MO, USA). Normal glucose (100 mg/dL) Dulbecco’s Modified Eagle Medium (DMEM; Catalog No.: 12320), penicillin/streptomycin (P/S), fetal bovine serum (FBS), horse serum, and trypsin-EDTA were bought from Invitrogen (Carlsbad, CA, USA). Rosiglitazone used in this experiment did not contain any inactive ingredients, and the pure compound was kindly provided by GlaxoSmithKline, Ltd. (Taipei, Taiwan). Phospho-AMPK, AMPK, phospho-ACC, ACC, phospho-AKT, AKT, PPARγ, C/EBPα, C/EBPβ, and GAPDH antibodies were purchased from Cell Signaling Technology (Danvers, MA, USA).
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3
Isolation and Differentiation of Preadipocytes
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Primary ING preadipocytes were isolated from 20-month old female GHR-/- and wild-type littermates as previously described [68 (link)]. In brief, ING WAT was minced, digested with collagenase type-2 (Worthington Biochemical Corp., Lakewood, NJ, 1 mg/ml HBBS, Gibco,Grand Island, NY), filtered through a 100 μm nylon mesh, and centrifuged at 1000 x g for 10 minutes. The pellets were resuspended in αMEM containing 10% NBCS (Gibco) and 1% penicillin-streptomycin (Gibco). Cells were then plated and maintained in a humidified incubator for 16 hours with 3% O2 and 5% CO2 before being washed, trypsinized, and replated at a density of 5x104 cells/cm2. This procedure results in over 90% pure preadipocyte populations as determined by morphology and assay of markers by RT-PCR [21 (link), 69 (link)]. Differentiation of preadipocyte cultures was induced by exposure to differentiation medium containing DMEM/F-12 (Gibco), 10% FBS (Gibco), 1 μg/ml bovine insulin (Sigma-Aldrich), 250 nM dexamethasone (Sigma-Aldrich), 0.5 mM IBMX (Sigma-Aldrich), and 2.5 μM rosiglitazone (GlaxoSmithKline, Philadelphia, PA). Cells were differentiated for 48 hours in a humidified incubator with 20% O2 and 5% CO2. Cultures were then visually analyzed for lipid droplet formation prior to being lysed for transcriptional analysis of adipogenesis markers.
4
Sourcing and Characterizing Pharmaceutical Compounds
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Digoxin, 95% and (+/−)-verapamil, 99% were obtained from Acros Organics (Morris Plains, NJ, USA); indomethacin, 98% and ranitidine hydrochloride were purchased from Alfa Aesar GmbH and Co KG (Karlsruhe, Germany), whereas losartan was obtained from Cayman Chemical (Ann Arbor, MI, USA). Atropine sodium salt, cefadroxil, fluvastatin natrium and valsartan were purchased from Sigma-Aldrich (St. Louis, MO, USA); rosuvastatin calcium salt was obtained from Santa Cruz Biotechnology Inc. (Dallas, TX, USA). Fexofenadine and sulfasalazine were purchased from Fluorochem Ltd. (Hadfield, UK), and rosiglitazone was obtained from GlaxoSmithKline (Brentford, UK). Agar, calcium chloride dihydrate, glucose hydrate, magnesium chloride hexahydrate, potassium chloride, sodium chloride, sodium phosphate monobasic and sodium hydrogen carbonate were obtained from Hänseler AG (Herisau, Switzerland).
5
Endothelin Receptor Antagonist Effects
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Male Sprague-Dawley rats were used and the experiments were conducted in accordance with the National Institutes of Health (NIH) Guide for the Care and Use of Laboratory Animals with the approval by the institutional committee. Polyclonal rabbit anti-ETBR antibody was from Abcam. Polyclonal rabbit anti-ETAR was from Santa Cruz Biotechnology. ET-1 and NG-nitro-L-arginine methyl ester (L-NAME) were from Sigma-Aldrich Co., rosiglitazone was from GlaxoSmithKline, and A192621 was from Abbott Laboratories.
6
Allergy Assay with ELISA and Imaging
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ELISA kit (Wuhan Huamei Bio-engineering Co., Ltd.); rosiglitazone (purchased from GlaxoSmithKline); aluminum potassium sulfate (purchased from Shanghai SANGON Biological Engineering Technology Service Co., Ltd); ovalbumin (OVA, Sigma); glycogen stain (PAS) kit (purchased from Fuzhou Maixin Biotechnology Development Co., Ltd.); ELISA kit (OVAs IgE, IL-4, and IL-13) were purchased from Wuhan Huamei Bio-engineering Co., Ltd.); 402AI type ultrasonic atomizer (purchased from Jiangsu Yuyue Medical Equipment & Supply Co., Ltd.); Microplate reader (purchased from USA BioTek Company); inverted microscope (purchased from Olympus company in Japan).
7
Investigating PGC-1α in SOD1(G93A) Mice
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Four-week-old SOD1(G93A) mice (body weight about 75 g) were purchased from the Nanjing biomedical research institute. C57BL/6 mice were purchased from Yangzhou University. All animal protocols were reviewed and approved by the Institutional Animal Care and Usage Committee of The First Clinical College of Harbin Medical University. The mice were kept in a temperature-controlled (22–24°C) room with a 12-h light and 12-h dark cycle and allowed to customize to the new environment for a week. According to the different groups (10 mice in each group), injected PBS or rosiglitazone (GlaxoSmithKline, Philadelphia, PA, USA) (10 mg/kg body weight) with siControl or siPGC-1α through tail-vein twice a week for 8 weeks.
8
Evaluating Antidiabetic Effects of PJE
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After four weeks of high-fat diet feeding, the fasted blood glucose levels of the mice were tested, and the mice who were fasted blood glucose levels ≥ 11.2 mmol/L were determined as type 2 diabetes mellitus. Then, the mice were divided randomly into four groups (n = 10 for each group). Group 1, diabetic control mice fed with saline; Group 2, diabetic mice fed with 700 mg/kg PJE; Group 3, diabetic mice fed with 350 mg/kg PJE; Group 4, diabetic mice fed with 4 mg/kg Rosiglitazone (GlaxoSmithKline, Tianjing, China). Body weight was measured at 7th and 14th day. After continuous feeding for 14 days, blood samples were collected from tail vein in heparin-containing sample tubes. Then the samples were centrifuged at 4°C (6000 g, 10 min) to collect plasma which was stored at –20°C for use.
9
Differentiation of Human Subcutaneous Preadipocytes
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Human white preadipocytes derived from subcutaneous adipose tissue of a female Caucasian subject (BMI 21 kg/m2; age 44 yr) were obtained from PromoCell (Heidelberg, Germany). Cells were cultured in preadipocyte growth medium supplemented with 100 U/ml penicillin, 100 μg/ml streptomycin, and 0.25 μg/ml amphotericin B (Lonza, Tewkesbury, UK) at 37°C in a humidified atmosphere of 5% CO2-95% air. Preadipocytes were seeded at 40,000/cm2 and grown in 6- or 24-well plates until confluence. At confluence, cells were induced to differentiate (day 0) by incubation for 3 days in Dulbecco's modified Eagle's medium (DMEM)-Ham's F-12 (1:1) medium containing 32 μM biotin, 1 μM dexamethasone, 200 μM 3-isobutyl-1-methylxanthine, 100 nM insulin, 11 nM l -thyroxine (all from Sigma, Poole, Dorset, UK), 8 μM rosiglitazone (GlaxoSmithKline, Uxbridge, UK), and 100 U/ml penicillin, 100 μg/ml streptomycin, and 0.25 μg/ml amphotericin B. After induction, cells were cultured in maintenance medium containing 3% fetal calf serum (FCS, Sigma), 100 nM insulin, 32 μM biotin, and 1 μM dexamethasone until full differentiation into adipocytes.
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