The experimental protocols are shown in Fig 1 . After a stabilization period of 20 minutes, the control hearts were perfused for another 10 minutes before ischemia-reperfusion in order to measure the baseline pre-ischemia cardiac function. Subsequently, global ischemia was applied by eliminating flow for 20 minutes followed by 40 minutes of reperfusion. In the phlorizin-perfused group, 10−4 mol/L of phlorizin (Sigma-Aldrich, Tokyo, Japan) was added to the buffer during the 10-minute pre-ischemia perfusion and 40-minute reperfusion periods. phlorizin was dissolved in dimethylsulfoxide (DMSO), and the solvent concentration was identically maintained in the control group. For the immunoblotting analysis, the individual perfused hearts after ischemia-reperfusion or the hearts simply perfused for an equivalent interval of the ischemia-reperfusion protocol (namely, 90 minutes) were snap frozen in liquid nitrogen and stored at -80°C prior to protein extraction. Triphenyltetrazolium chloride (TTC) staining was performed to determine the myocardial infarct size using the individual perfused hearts after ischemia-reperfusion, as previously described [3 (link)].
Phlorizin
Manufactured by Merck Group
Sourced in United States, Japan
Phlorizin is a chemical compound that acts as a sodium-glucose co-transporter 2 (SGLT2) inhibitor. It is a naturally occurring substance found in the bark of apple trees. Phlorizin is used as a research tool in the study of glucose metabolism and transport processes.
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Lab products found in correlation
Phloretin, by Merck Group (9 mentions)
Quercetin, by Merck Group (6 mentions)
Rutin, by Merck Group (5 mentions)
Caffeic acid, by Merck Group (4 mentions)
Ferulic acid, by Merck Group (4 mentions)
Trolox, by Merck Group (4 mentions)
P coumaric acid, by Merck Group (4 mentions)
Gallic acid, by Merck Group (3 mentions)
Sodium carbonate, by Kemika (3 mentions)
Caffeic, by Merck Group (3 mentions)
Chlorogenic, by Merck Group (3 mentions)
2 4 6 tri 2 pyridyl s triazine tptz, by Thermo Fisher Scientific (3 mentions)
Copper 2 chloride, by Thermo Fisher Scientific (3 mentions)
P coumaric, by Merck Group (3 mentions)
Potassium persulfate, by Kemika (3 mentions)
Neocuproine, by Thermo Fisher Scientific (3 mentions)
Cyanidin 3 glucoside, by Extrasynthese (3 mentions)
Chlorogenic acid, by Merck Group (3 mentions)
Kaempferol, by Merck Group (3 mentions)
Catechin, by Merck Group (3 mentions)
25 protocols using phlorizin
1
Phlorizin Protects Against Ischemia-Reperfusion Injury
2
Diabetes Mellitus Experiment in Rats
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Based on the biological parameters, SD rats and SDT fatty rats at 4 weeks of age were divided into four subgroups; Group 1 (normal control): SD rats subcutaneously injected with 20 % propylene glycol and fed powdered CRF-1 chow; Group 2 (DME vehicle): SDT fatty rats subcutaneously injected with 20 % propylene glycol and fed powdered CRF-1 chow; Group 3 (DME pioglitazone): SDT fatty rats subcutaneously injected with 20 % propylene glycol and administered 10 mg/kg/day pioglitazone (synthesized at Japan Tobacco Co.) through powdered CRF-1 chow (food admixture); and Group 4 (DME phlorizin): SDT fatty rats subcutaneously injected with 150 mg/kg/day phlorizin (Sigma-Aldrich Co., St. Louis, MO, USA) and fed powdered CRF-1 chow.
3
Antibodies for Cellular Analysis
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Unless otherwise specified, reagents including phlorizin (phlorizin, P3449) were obtained from Sigma-Aldrich. The antibodies that we used in this study were as follows: p63 antibody (#sc-8431, Santa Cruz Biotechnology, Santa Cruz, CA), proliferating cell nuclear antigen antibody (PCNA, #M0879, DAKO, Glostrup, Denmark), integrin α6 antibody (#sc-6597, Santa Cruz Biotechnology), integrin β1 antibody (#sc-9970, Santa Cruz Biotechnology), and type IV collagen antibody (239M-16, Cell Marque, Rocklin, CA).
4
Phlorizin Treatment in Diabetic Mice
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Phlorizin (Sigma-Aldrich) was prepared as a 20% stock (0.2 g ml−1) in 1,2-propanediol (Sigma-Aldrich) and kept at 4 °C. Mice were injected subcutaneous with 0.4 g kg−1 Phlorizin or vehicle once daily for 7 days (10-week-old db/db; 3 phlorozin-treated and 7 vehicle-treated mice) or 10 days (4-week-old db/db; 3 phlorozin-treated and 7 vehicle-treated mice). Non-fasted blood glucose was measured before killing. For the Phlorizin-treated 4-week-old db/db mice it was 4.7±0.4 mM and for the control-treated 4-week-old db/db mice it was 4.8±0.6 mM. For the Phlorizin-treated 10-week-old db/db mice it was 8.4±1.5 mM and for the control-treated 10-week-old db/db mice it was 20.0±3.2 mM. Animals were killed by cervical dislocation and pancreatic islets and RNA was prepared as described above.
5
Microbial Cultivation and Phytochemical Analysis
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De Man, Rogosa, Sharpe (MRS) broth and Esculin-MRS agar medium was purchased from Sharing Technologies (Shanghai, China). Potassium ferricianyde, ferric chloride, ferrous sulfate, trichloroacetic acid, salicylic acid, and methanol (High Performance Liquid Chromatography, HPLC grade) were purchased from Tianli Biochemical Co., Ltd. (Tianjin, China). 2,2-diphenyl-1-picrylhydrazyl (DPPH) and Folin–Ciocalteus reagent were purchased from Biotopped Life Sciences (Beijing, China). Gallic acid, epicatechin acid, ferulic acid, caffeic acid, quercitrin, quercetin, phlorizin, phloretin, and p-nitrophenyl-β-D-glucopyranoside were purchased from Sigma-Aldrich (Beijing, China). All other reagents were purchased from Basifu Co., Ltd. (Tianjin, China).
6
Antioxidant and Anti-Diabetic Potential of Pectin and Apple Fiber
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The apple fiber powder was supplied by Biesterfeld AG (Zagreb, Croatia) and both types of pectin were the product of CP Kelco (Atlanta, GA, USA). The 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS), trolox, 4-(dimethylamino)-cinnamaldehyde (DMAC), chlorogenic, caffeic, p-coumaric, gallic and ellagic acids, rutin, quercetin, phloretin and phlorizin were purchased from Sigma-Aldrich (St. Louis, MO, USA). The α-glucosidase (from Saccharomyces cerevisiae) and acarbose were obtained from Sigma-Aldrich (St. Louis, MO, USA). The 4-nitrophenyl-α-D-glucopyranoside was obtained from Alfa Aesar (Kandel, Germany) and potassium dihydrogen phosphate was from BDH Prolabo (Poole, UK). Potassium persulfate, Folin-Chiocalteu reagent and sodium carbonate were obtained from Kemika (Zagreb, Croatia). Neocuproine, 2,4,6-tri(2-pyridyl)-s-triazine (TPTZ) and copper (II) chloride were purchased from Acros Organics (Geel, Belgium). Methanol (HPLC grade) was from J.T. Baker (Deventer, The Netherlands) and orthophosphoric acid (HPLC grade > 85%) was from Fisher Scientific (Loughborough, UK). Iron (III) chloride hexahydrate, sodium acetate, ethanol, ammonium acetate and starch were bought from Gram-mol (Zagreb, Croatia). Cyanidin 3-glucoside was from Extrasynthese (Genay, France).
7
Phlorizin Enhances Endothelial Function
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Phlorizin was obtained from Sigma-Aldrich (St Louis, MO). HUVECs were obtained from Shanghai Institute of Biochemistry and Cell Biology (Shanghai, China). LY294002 was purchased from Calbiochem (CA, USA). Antibodies to anti-phosphor-AKT (S473), anti-phosphor-eNOS (S1177), and anti-phosphor-IRS-1 were purchased from Cell Signaling Technology (Danvers, MA). Anti-SGLT1 was purchased from Affinity (Cincinnati, OH), and anti-SGLT2 was purchased from Abcam (Cambridge, UK). The nitric oxide assay kit was obtained from Cayman Chemical (Ann Arbor, MI).
8
Fluorescent Glucose Derivative Synthesis
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9
Comprehensive Phytochemical Analysis of Apple Fibers
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Apple fibers were obtained from Biesterfeld AG (Zagreb, Croatia). 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS), 4-(dimethylamino)-cinnamaldehyde (DMAC), trolox, 2,2-diphenyl-1-picrylhydrazyl (DPPH), rutin, quercetin, phloretin, phlorizin, chlorogenic, caffeic, p-coumaric, gallic, ellagic acids, myrtenol and α-amylase (from the porcine pancreas) were obtained from Sigma-Aldrich (St. Louis, MO, USA). Potassium persulfate, Folin–Ciocalteu reagent and sodium carbonate were purchased from Kemika (Zagreb, Croatia). Neocuproine, 2,4,6-tri(2-pyridyl)-s-triazine (TPTZ) and copper (II) chloride were bought from Acros Organics (Geel, Belgium). Methanol (HPLC grade) was from J.T. Baker (Deventer, Netherlands) and orthophosphoric acid (HPLC grade > 85%) was from Fisher Scientific (Loughborough, UK). Iron (III) chloride hexahydrate, sodium acetate, ethanol, ammonium acetate and starch were bought from Gram-mol (Zagreb, Croatia). Cyanidin 3-glucoside was from Extrasynthese (Genay, France) and 3,5-dinitrosalicylic acid from Alfa Aesar (Kandel, Germany).
10
Glucose Uptake Mechanism Evaluation
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2-NBDLG (23003-v, Peptide Institute), 2-NBDG (23002-v, Peptide Institute), 2-TRLG (Peptide Institute), phloretin (P7912, Sigma), cytochalasin B (C6762, Sigma), phlorizin (P3449, Sigma) and 4′,6-diamidino-2-phenylindole (DAPI, 049-18801, Wako) were prepared as described previously [18 (link)]. cytochalasin B was used to examine an involvement of GLUTs in the uptake of glucose tracers, since it acts as a specific antagonist of GLUTs when applied briefly in a low dose [11 (link)]. phlorizin was used to inhibit SGLTs [2 (link)]. In all experiments, 100 μM of carbenoxolone (C4790, Sigma) was routinely added to exclude non-specific permeation of the fluorescent tracers through gap junctions/hemichannels [28 (link)]. d -glucose and l -glucose were purchased from Fujifilm Wako Pure Chemical (049-31165) and Tokyo Chemical Industry (921-60-8), respectively.
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