Streptozotocin (STZ) and MRS 1730 were purchased from Tocris Cookson. The anti-iNOS antibody was from BD Biosciences, whereas the peroxidase-coupled secondary antibody was obtained from Vector. Recombinant rat cytokines were obtained from Tebu-Bio. Unless otherwise specified, chemicals were purchased from Sigma-Aldrich.
Streptozotocin (stz)
Manufactured by Bio-Techne
Sourced in United Kingdom
STZ is a lab equipment product manufactured by Bio-Techne. It is a chemical compound used for inducing diabetes in experimental animal models.
Automatically generated - may contain errors
Lab products found in correlation
Streptozotocin (stz), by Roche (3 mentions)
Glucometer, by Roche (2 mentions)
Diazepam, by Merck Group (2 mentions)
Loganin, by Extrasynthese (2 mentions)
Accu check performa, by Roche (2 mentions)
Anti inos antibody, by BD (1 mentions)
Complete freund adjuvant (cfa), by Merck Group (1 mentions)
Complete freund s adjuvant, by Merck Group (1 mentions)
Win55 212, by Bio-Techne (1 mentions)
Hu210, by Bio-Techne (1 mentions)
Accu chek active kit, by Roche (1 mentions)
Cd 1 mice, by Charles River Laboratories (1 mentions)
8 protocols using streptozotocin (stz)
1
Streptozotocin-Induced Diabetic Model
2
Diabetic Rats' Behavioral Response
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Seven days after adaptation, rats were randomly divided into five groups (n = 8): control (con), diabetic plus saline (dia + saline), diabetic plus imipramine (dia + imipr), diabetic plus diazepam (dia + diaz), and diabetic plus loganin (dia + log). Experimental diabetes was induced by intraperitoneal (i.p.) injection of a single dose of 60 mg/kg streptozotocin (STZ) (Tocris, UK) (35) . Fasting blood glucose levels were determined 72 h after STZ injection, using a glucometer (Roche, Germany). Blood samples were collected from the tail vein of overnight (10 h) fasted rats. Rats were considered diabetic, if blood glucose levels exceeded 300 mg/dl. Confirming diabetes (on day 4 following STZ injection), rats received saline (1 ml), imipramine (15 mg/kg i.p.) (ScienceLab, USA) (7) , or diazepam (1 mg/kg i.p.) (Sigma, USA), 30 min before behavioral tests (14) . The rats in the dia + log group received 40 mg/kg of loganin per os (Extrasynthese, France), which was administered at 9:00 a.m. for 10 consecutive days. On day 10, the behavioral tests were performed an hour after the loganin administration.
3
Diabetic Rats' Behavioral Response
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Seven days after adaptation, rats were randomly divided into five groups (n = 8): control (con), diabetic plus saline (dia + saline), diabetic plus imipramine (dia + imipr), diabetic plus diazepam (dia + diaz), and diabetic plus loganin (dia + log). Experimental diabetes was induced by intraperitoneal (i.p.) injection of a single dose of 60 mg/kg streptozotocin (STZ) (Tocris, UK) (35) . Fasting blood glucose levels were determined 72 h after STZ injection, using a glucometer (Roche, Germany). Blood samples were collected from the tail vein of overnight (10 h) fasted rats. Rats were considered diabetic, if blood glucose levels exceeded 300 mg/dl. Confirming diabetes (on day 4 following STZ injection), rats received saline (1 ml), imipramine (15 mg/kg i.p.) (ScienceLab, USA) (7) , or diazepam (1 mg/kg i.p.) (Sigma, USA), 30 min before behavioral tests (14) . The rats in the dia + log group received 40 mg/kg of loganin per os (Extrasynthese, France), which was administered at 9:00 a.m. for 10 consecutive days. On day 10, the behavioral tests were performed an hour after the loganin administration.
4
Diabetes Induction in Animal Models
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In order to induce diabetes, animals received intraperitoneal (i.p.) injections of STZ (35 mg/kg, Tocris Bioscience, UK) or vehicle (0.1M citrate buffer pH 4.6) under general anesthesia with isoflurane inhalation. Body weight, blood glucose levels, and mechanical allodynia were measured before the injections and weekly after that. Diabetes was confirmed by assaying the glucose levels in blood obtained from the tail vein using Accu-Check Performa (Roche, Germany). Rats with glucose levels greater than 200 mg/dl 3 days after STZ injection were considered diabetic.
5
Induction and Confirmation of Diabetes in Rats
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To induce diabetes, rats received intraperitoneal (I.P.) injections of STZ (35 mg/kg, Tocris Bioscience, Bristol, UK) or vehicle (0.1 M citrate buffer pH 4.6). These injections were performed under brief isoflurane anesthesia. Baseline reading of mechanical allodynia, blood glucose concentrations and body weight were taken before the injections, and then tested every week for 8 weeks [23 (link)]. In order to confirm diabetes, Accu-Check performa (Roche, Grenzach-Wyhlen, Germany) was used to measure the glucose concentration in blood samples collected from the tail vein. Three days following the STZ injections, rats with glucose levels higher than 200 mg/dl were regarded as diabetic.
6
Pharmacological Modulation of Pain Pathways
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HU210, WIN55212 and STZ (streptozotocin) were purchased from Tocris Bioscience (Abingdon, UK). CFA (Complete Freund’s Adjuvant) was purchased from Sigma-Aldrich (Munich, Germany). Tramadol was purchased from The Arab Pharmaceutical Manufacturing Co. Ltd. (Amman, Jordan). Morphine was purchased from Hikma Pharmaceuticals LLC (Amman, Jordan). Unless otherwise indicated, drugs were initially dissolved in ethanol (100%) to form a stock solution, and then diluted in (3% tween 20 in saline).
7
Streptozotocin-Induced Diabetic Mice Model
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Six-week-old male C57BL/6 mice were used for the experiment. All animal procedures were conducted per the Institutional Animal Care and Use Committee of CHA University (IACUC No: 170028). Mice were allowed to acclimate to the environment of a well-ventilated room for a week prior to the experiment. They were fasted for 12–14 h before intraperitoneal injection of STZ (Tocris, Bristol, UK) in 100 mM citrate buffer (pH 4.5). Blood glucose levels after STZ treatment were measured every three to four days with the Accu-Chek active kit (Roche, Basel, Switzerland). Mice with a blood glucose level higher than 250 mg/dL are generally used for the diabetic mouse model [27 (link)]. In the present study, a significant rise in the blood glucose level was observed two weeks after two daily STZ treatments at 100 mg/kg. Mice with blood glucose level higher than 250 mg/dL were selected and used for wound healing assay.
8
Hyperglycemic Mouse Model for SGLT2i Evaluation
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Mouse experiments were approved by the Joslin Diabetes Center Institutional Animal Care and Use Committee. Male 8-week-old CD-1 mice (strain IGS, stock no. 022) were purchased from Charles River Laboratories. Mice were group housed in a pathogen-free facility and maintained on a 12-h normal light/dark cycle with ad libitum access to diet. All mice consumed a low-fat diet (Research Diets, D14020502) consisting of 17% kcal from protein, 73% carbohydrate, and 10% fat. To induce hyperglycemia, STZ (Tocris Bioscience, cat. no. 1621,) was dissolved in citrate buffer (114 mmol/L) and administered in 40 mg/kg intraperitoneal injections on 2 consecutive days after a 4-h fast. Control (Con) mice were injected with citrate buffer vehicle. Two weeks after injection, a subset of STZ-treated mice was randomized to receive the SGLT2i Cana in their diet. For Cana treatment, the D14020502 diet was supplemented with lyophilized Cana (240 mg/kg diet; Advanced ChemBlocks, cat. no. 10300). Mice consumed ∼4–5 g of food per day, resulting in a Cana dose of ∼30 mg/kg/day. This approach was previously reported to successfully reduce blood glucose concentrations in hyperglycemic mice (22 ).
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