The cultured atrial fibroblasts were randomly divided into seven groups and treated as follows: i) NC group, cells were cultured in DMEM medium containing 4 mM glucose without any treatment; ii) Glu group, cells were cultured in DMEM medium containing 25 mM glucose without other treatment; iii) Glu+Apo group, cells were cultured in DMEM medium containing 25 mM glucose and 100 µg/ml apocynin (Sigma-Aldrich; Merck KGaA); iv) H2O2 group, cells were cultured in DMEM medium containing 4 mM glucose and treated with 100 nmol/l H2O2; v) H2O2+Apo group, cells were cultured in DMEM medium containing 4 mM glucose and 100 µg/ml apocynin, and treated with 100 nmol/l H2O2; vi) Glu+H2O2 group, cells were cultured in DMEM medium containing 25 mM glucose and treated with 100 nmol/l H2O2; vii) Glu+H2O2+Apo group, cells were cultured in DMEM medium containing 25 mM glucose and 100 µg/ml apocynin, and treated with 100 nmol/l H2O2.
Apocynin
Manufactured by Merck Group
Sourced in United States, Germany, Italy, United Kingdom, Sao Tome and Principe, Macao, Brazil, China, France, Austria
Apocynin is a chemical compound used in laboratory settings. It functions as an inhibitor of the enzyme NADPH oxidase, which plays a role in the production of reactive oxygen species. Apocynin is commonly utilized in research applications to investigate the involvement of oxidative stress in various biological processes.
Automatically generated - may contain errors
Lab products found in correlation
Tempol, by Merck Group (17 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (17 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (14 mentions)
Catalase, by Merck Group (11 mentions)
Acetylcholine, by Merck Group (11 mentions)
Ang 2, by Merck Group (9 mentions)
Allopurinol, by Merck Group (9 mentions)
N acetylcysteine, by Merck Group (9 mentions)
Sodium nitroprusside, by Merck Group (8 mentions)
Rotenone, by Merck Group (8 mentions)
N acetyl l cysteine, by Merck Group (8 mentions)
Phenylephrine, by Merck Group (7 mentions)
Losartan, by Merck Group (7 mentions)
L name, by Merck Group (7 mentions)
Indomethacin, by Merck Group (7 mentions)
N acetylcysteine nac, by Merck Group (7 mentions)
Lucigenin, by Merck Group (7 mentions)
Sb203580, by Merck Group (6 mentions)
Streptomycin, by Merck Group (6 mentions)
N acetyl cysteine (nac), by Merck Group (6 mentions)
243 protocols using apocynin
1
Atrial Fibroblast Oxidative Stress Response
2
Stimulants Elicit Cardiorespiratory Reflexes
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To stimulate the LVCFs, capsaicin [1 μg/kg; a transient receptor potential vanilloid 1 (TRPV1) receptor agonist] (Lin et al., 2013 (link)), adenosine (an adenosine A1-receptor agonist; 200 μg/kg, Sigma) (Lin et al., 2009 (link)), and α,β-methylene-ATP (15 μg/kg in the reflex studies and 100 μg/kg in the electrophysiological studies; a P2X receptor agonist) (Lin et al., 2013 (link)) were injected as a bolus into the right atrium (volume 0.1 ml) and then flushed by an injection of 0.3 ml of saline. MnTMPyP [manganese (III) tetrakis(1-methyl-4-pyridyl)porphyrin; a potent scavenger of superoxide anions; 5 mg/kg/day; Calbiochem, San Diego, CA] and apocynin (an inhibitor of NADPH oxidase; 30 mg/kg/day, Sigma) were given daily by intraperitoneal injection (volume ~0.35 ml) at 15 min prior to CIH or RA exposure for 14 consecutive days. The vehicle for adenosine, α,β-methylene-ATP, and MnTMPyP was saline. The vehicle for capsaicin was a solution that contained 10% Tween 80, 10% ethanol, and 80% saline. apocynin was prepared by dissolving the agent in 20% dimethyl sulfoxide (Sigma) and then diluting the resulting solution with saline.
3
Preparation of Chemical Reagents for Cell Assays
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LPS (Invivogen, Thermo Fisher Scientific, Carlsbad, CA, USA) and 2-DG (Sigma-Aldrich, St. Louis, MO, USA, Cat no. D8375) were dissolved in H2O at a concentration of 1 mg/mL and 1 M, respectively. 6-aminonicotinamide (6-AN; Sigma-Aldrich, St. Louis, MO, USA, Cat no. A68203) was dissolved in dimethyl sulfoxide (DMSO; Sigma-Aldrich, St Louis, MO, USA, cat. no. 472301; 100%) at a concentration of 200 mM. L-012 (WAKO Chemicals, Richmond, MO, USA, cat. no. 120-04891) and phorbol 12,13-dibutyrate (PDB; Sigma-Aldrich, St Louis, MO, USA, cat. no. P1269-5MG) were dissolved in DMSO in 10 μL and 5 μL aliquots, respectively, at a concentration of 1 × 10−2 M. IFN antibody receptor 1 (IFNAR; Bio X cell, Lebanon, NH, USA, Cat no BE0241) was diluted 1:1000 in cell culture medium. Apocynin (Sigma-Aldrich, St. Louis, MO, USA, Cat no. A10809) was dissolved in DMSO at a concentration of 300 mM. Chemicals were stored at −20 °C and rapidly thawed when required, apart from 6-AN and Apocynin, which were freshly prepared before each use.
4
PC12 Cell Glucose Toxicity Mitigation
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PC12 cells were supplied by Pasteur Institute of Iran (Tehran, Iran). Cells were kept at 37 °C and 90 % air humidity with 5 % CO2. Cells were cultured in DMEM with 5 % (v/v) FBS, 10 % (v/v) HS, 100 units/ml penicillin, and 100 μg/ml streptomycin. The culture medium was replaced every 48 h and the cells were passaged every 2–3 days.
The glucose concentration in DMEM was 25 mM and considered the normal glucose. The cells were used after 2 subcultures. In MTT assay, HG condition was estimated by exposing 96 well-culture plate (5000 cells/well) to 60 mM, 80 mM, 100 mM, and 140 mM D-glucose. Also, the cells were pretreated by Atorvastatin (0.001–1 μM) for 96 h and co-incubated with Atorvastatin and HG (140 mM) for 24 h. In experiments where Apocynin (Sigma, a10809) was used together with HG, the cells were pretreated with Apocynin (10, 20, 40 μm) 45 min before glucose addition. Control cells were cultured in normal glucose (25 mM). Atorvastatin was dissolved in dimethyl sulfoxide (DMSO). The DMSO′s final concentration was less than 0.1 %.
The glucose concentration in DMEM was 25 mM and considered the normal glucose. The cells were used after 2 subcultures. In MTT assay, HG condition was estimated by exposing 96 well-culture plate (5000 cells/well) to 60 mM, 80 mM, 100 mM, and 140 mM D-glucose. Also, the cells were pretreated by Atorvastatin (0.001–1 μM) for 96 h and co-incubated with Atorvastatin and HG (140 mM) for 24 h. In experiments where Apocynin (Sigma, a10809) was used together with HG, the cells were pretreated with Apocynin (10, 20, 40 μm) 45 min before glucose addition. Control cells were cultured in normal glucose (25 mM). Atorvastatin was dissolved in dimethyl sulfoxide (DMSO). The DMSO′s final concentration was less than 0.1 %.
5
Apocynin Ameliorates CRF in Rats
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Animal experiments were approved by the Committee on Ethics of Animal Experiments, Sun Yat-sen University and conducted in accordance with the Guidelines for the Care and Use of Laboratory Animals published by the US National Institutes of Health (NIH Publication No. 85–23, revised 1996).
Male Sprague-Dawley (SD) rats weighing 160–180g were obtained from Sun Yat-sen University. A two-step 5/6 NE model of CRF was used as described in our previous study [28 (link)]. Rats were randomly divided into three groups: sham-operated group, NE group and NE+apocynin group. Rats in NE+apocynin group were fed with 1.5 mM apocynin (Sigma-Aldrich, St. Louis, US) in drinking water [30 (link)]. 8 weeks after surgery, SBP, DBP and heart rate of the animals were measured by a tail-cuff method (BP-98A, Softron, Tokyo, Japan). Plasma was obtained by tail vein at 8 weeks for determining the levels of creatinine (model 7600–010, HITACHI automatic analyzer, Tokyo, Japan) and 14, 15-EET levels (Detroit R&D Inc.). Cardiac tissues were harvested at sacrifice for histological and molecular investigations.
Male Sprague-Dawley (SD) rats weighing 160–180g were obtained from Sun Yat-sen University. A two-step 5/6 NE model of CRF was used as described in our previous study [28 (link)]. Rats were randomly divided into three groups: sham-operated group, NE group and NE+apocynin group. Rats in NE+apocynin group were fed with 1.5 mM apocynin (Sigma-Aldrich, St. Louis, US) in drinking water [30 (link)]. 8 weeks after surgery, SBP, DBP and heart rate of the animals were measured by a tail-cuff method (BP-98A, Softron, Tokyo, Japan). Plasma was obtained by tail vein at 8 weeks for determining the levels of creatinine (model 7600–010, HITACHI automatic analyzer, Tokyo, Japan) and 14, 15-EET levels (Detroit R&D Inc.). Cardiac tissues were harvested at sacrifice for histological and molecular investigations.
6
Investigating Angiotensin II-induced H9c2 Cell Responses
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The H9c2 rat cell line was obtained from American Type Culture Collection (ATCC) (Sanger Biotech, Shang Hai, China). Cells were cultured in standard Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% fetal bovine serum in a humidified incubator with 5% CO2 at 37°C. Cells were cultured in serum-free DMEM for 24 h before treatment. Four groups were divided: (a) dimethyl sulfoxide (DMSO) alone, (1 μl, Sigma-Aldrich, St. Louis, US) (b) apocynin (100 μM) alone (c) Ang II (100 nM, Sigma-Aldrich, St. Louis, US) (d) Ang II (100 nM)+apocynin (100 μM).
7
Osteoarthritis Induction and Treatment
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30C57bl/6 mice (initial weight 18–20g, Animal Experiment Center, Southern Medical University, Guangzhou, China) were used in this study. All the mice were housed under a regulate light/dark cycle with food and water available ad libitum. They were acclimated for 7 days before any experimental procedures. All the animal experimental procedures were approved by the Laboratory Animal Care and Use Committee of Nanfang Hospital, Southern Medical University (NFYY-2015-68).
Osteoarthritis mice models were established by knee anterior cruciate ligament transaction (ACLT). Mice were randomized into five groups: Group 1, sham-operated; Group 2, ACLT-operated and treated with PBS (pH = 7.4); Group 3, ACLT-operated and treated with AOPPs (50 mg/kg/d); Group 4, intraperitoneal injection of AOPPs-MSA (50 mg/kg/d) and intragastric administration of apocynin (a nicotinamide adenine dinucleotide phosphate (NADPH) oxidase inhibitor, Sigma, USA) at 100 mg/kg/d dissolved in Tween-80; Group 5, intragastric administration of apocynin at 50 mg/kg/d dissolved in Tween-80.
Osteoarthritis mice models were established by knee anterior cruciate ligament transaction (ACLT). Mice were randomized into five groups: Group 1, sham-operated; Group 2, ACLT-operated and treated with PBS (pH = 7.4); Group 3, ACLT-operated and treated with AOPPs (50 mg/kg/d); Group 4, intraperitoneal injection of AOPPs-MSA (50 mg/kg/d) and intragastric administration of apocynin (a nicotinamide adenine dinucleotide phosphate (NADPH) oxidase inhibitor, Sigma, USA) at 100 mg/kg/d dissolved in Tween-80; Group 5, intragastric administration of apocynin at 50 mg/kg/d dissolved in Tween-80.
8
Apocynin Inhibits NOX-Driven Oxidative Stress
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To observe the effect of inhibition of NOX enzyme on oxidative stress, SH-SY5Y cells were treated with apocynin (sigma), a NOX inhibitor, for 24 h. The concentration of apocynin used in this study is 10−4 mol/L. Following that, the cells were sampled for relevant analysis, such as gene expression measurement and other assays.
9
Antioxidants Modulate Cardiomyocyte Oxidative Stress
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H9C2 cardiomyocytes were incubated with N-acetyl-cysteine (NAC) (Sigma-Aldrich, St. Louis, MO, USA) at 2 mM or tempol (Sigma-Aldrich, St. Louis, MO, USA) at 10 mM for 6 h. PBS containing 0.1 % bovine serum albumin was used as a control in all protocols. The dosage for NAC and tempol used in this study was based on the previous studies (Peng et al. 2011 , Rahman et al. 2014) . NAC is a precursor of the cellular antioxidant glutathione. tempol is a membrane-permeable piperidine nitroxide that mimics SODs by dismuting O 2 -anions.
tempol also detoxifies redox-reactive transition metal ions and directly reacts with many ROS forming adducts.
NADPH oxidase inhibitor apocynin treatment H9C2 cardiomyocytes were treated with apocynin (100 μmol/l) (Sigma-Aldrich, St. Louis, MO, USA) for 6 h. DMSO (0.02 %) was used as control. The dose used in this study was based on the previous report (Qin et al. 2007) .
tempol also detoxifies redox-reactive transition metal ions and directly reacts with many ROS forming adducts.
NADPH oxidase inhibitor apocynin treatment H9C2 cardiomyocytes were treated with apocynin (100 μmol/l) (Sigma-Aldrich, St. Louis, MO, USA) for 6 h. DMSO (0.02 %) was used as control. The dose used in this study was based on the previous report (Qin et al. 2007) .
10
Antioxidant and Oxidative Stress Assays
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Apocynin (C9H10O3, A10809, Figure 1A ), protocatechuic acid (C7H6O4, 37580, Figure 1B ), acetylcholine (A6625), phenylephrine (P6126), dihydroethidium (DHE), 4,5-diaminofluorescein diacetate (DAF-2DA), 2,2-Diphenyl-1-picrylhydrazyl (DPPH, D9132), 2,2′-Azobis (2-methylpropionamidine) dihydrochloride (AAPH, 440914) (±)-6-Hydroxy-2,5,7,8-tetramethylchromane-2-carboxylic acid (Trolox, 238,813), Brij 35 (801,962), Tung oil (440,337), and fluorescein (F6377) were acquired from Sigma Aldrich (United States). Lucigenin (L6868) was obtained from ThermoFisher Scientific (United States). DMEM was purchased from Vitrocell (00,025, Brazil) and FBS from Gibco (12657029, South America). TBARS assay kit was acquired from Cayman Chemical (10009055, United States). The remaining salts and reagents were acquired from Sigma Aldrich.
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