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5 protocols using β nicotinamide adenine dinucleotide phosphate nadph

1

Antioxidant and Nitric Oxide Evaluation

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Ellagic acid (purity ≥ 95%), L-NAME, ethylenediaminetetraacetic acid (EDTA), butylated hydroxytoluene (BHT), thiobarbituric acid (TBA), sodium dodecyl sulfate (SDS), sulfanilamide, N-1-napthylethylethylenediamine (NED) and 1,1,3,3-tetraethoxypropane were obtained from Sigma-Aldrich Corp. (St. Louis, MO, USA). β-Nicotinamide adenine dinucleotide phosphate (NADPH), glucose-6-phosphate dehydrogenase (G-6-PD) and Nitrate reductase were obtained from Roche Applied Sciences (Mannheim, Germany). Trichloroacetic acid (TCA) and lucigenin were obtained from Fluka Chemika Co., Ltd (Buch, Switzerland). All chemicals used in this study were obtained from standard companies and were of analytical grade quality.
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2

Characterization of Ganoderic Acid Metabolism

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GAA was purchased from Weikeqi Biotechnology Co, Ltd. (Chengdu, Sichuan province, China). Ganoderic acid B, ganoderic acid C2 (GAC2) and ganoderic acid D were kindly provided by professor Ruo-Yun Chen (Institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China). Glycyrrhizic acid (GLA), α-naphthoflavone, quinidine, fluconazole, diethyldithiocarbamate and uridine 5′-diphosphoglucuronic acid (UDPGA) were obtained from Sigma-Aldrich Chemical Co. (St. Louis, MO, USA). Ticlopidine and ketoconazole were purchased from the National Institutes for Food and Drug Control (Beijing, China). β-nicotinamide adenine dinucleotide phosphate (NADPH) was purchased from Roche Molecular Biochemicals (Basel, Switzerland). A pooled Sprague Dawley rat liver microsomes (RLMs) was obtained from RILD Research Institute for Liver Diseases Co. Ltd (Shanghai, China). A pooled human liver microsomes (HLMs) was obtained from BD Gentest Corporation (Woburn, MA, USA). Methanol and ethyl acetate of HPLC grade, and acetonitrile and formic acid of LC/MS grade, were obtained from Fisher Co. Ltd. (Waltham, MA, USA). Milli-Q (Millford, MA, USA) water was used throughout the study.
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3

Cytochrome P450 Enzyme Profiling

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BDQ and propranolol were purchased from Sigma-Aldrich (St. Louis, MO, United States); TBI-166 was provided by the Institute of Materia Medica, Peking Union Medical College and Chinese Academy of Medical Sciences (Beijing, China); N-desmethyl-bedaquilin (M2) were purchased from Toronto Research Chemicals Inc. (Toronto, Canada); Probe substrates and associated metabolites for CYPs were purchased from BD Bioscience (San Jose, CA, United States); β-nicotinamide adenine dinucleotide phosphate (NADPH) was purchased from Roche (Roche, Switzerland); dimethyl sulfoxide, formic acid, acetic acid were purchased from Sigma-Aldrich (St. Louis, MO, United States);, acetonitrile, and methanol were purchased from Fisher Co. Ltd. (Waltham, MA, United States); Human liver microsomes (HLM; 150-donor pool, mixed sex) and recombinant human cytochrome P450s (rCYPs; rCYP1A2, 2A6, 2B6, 2C8, 2C9, 2C19, 2D6, 2E1, 2J2, 3A4, 3A5, and 4F2) were purchased from BD Gentest (Woburn, MA, United States); Milli-Q (Millford, MA, United States) water was used throughout the study.
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4

Characterization of Cytochrome P450 Interactions

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RTV, LPV, and indinavir sulfate were purchased from Macklin Inc. (Shanghai, China); testosterone was purchased from Sigma-Aldrich (St. Louis, MO, United States); 6β-hydroxytestosterone was purchased from Toronto Research Chemicals Inc. (Toronto, Canada); β-nicotinamide adenine dinucleotide phosphate (NADPH) was purchased from Roche (Roche, Switzerland); propranolol was obtained from the National Institutes for Food and Drug Control (Beijing, China); Solutol HS-15 was purchased from MedChemExpress LLC (Shanghai, China); and XYPI was purchased from Jiangxi Qingfeng Pharmaceutical Co. Ltd. (Jiangxi, China). A pooled sample of human liver microsomes (HLM, 452161) was obtained from BD Gentest Corporation (BD Gentest TM, Woburn, MA, United States). Antibodies specific for CYP3A1 (Mouse monoclonal antibody) was purchased from Santa Cruz Biotechnology (Cruz, CA, United States). Primary antibody for GAPDH (Rabbit Polyclonal antibody) and the second-antibody specific for Rabbit and Mouse were purchased from Proteintech Group, Inc. Methanol and acetonitrile of high-performance liquid chromatography (HPLC) grade were obtained from Fisher Co. Ltd. (Waltham, MA, United States). Milli-Q (Millford, MA, United States) water was used throughout the study.
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5

Comprehensive Analytical Procedures for Pharmacokinetics

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Chlorzoxazone, dextromethorphan, tolbutamide, testosterone, omeprazole, 6-hydroxyChlorzoxazone, dextrorphan, and 5-hydroxyomeprazole were purchased from Sigma-Aldrich Company (St. Louis, USA). 6β-Hydroxytestosterone and 4-hydroxytolbutamide were purchased from Toronto Research Chemicals Inc. (Toronto, Canada). β-Nicotinamide adenine dinucleotide phosphate (NADPH) was purchased from Roche (Roche, Switzerland). Vinpocetine, propranolol, and gliclazide were obtained from the National Institutes for Food and Drug Control (Beijing, China). Acetonitrile of high-performance liquid chromatography grade was obtained from Fisher Co. Ltd. (Waltham, MA, USA). Milli-Q (Milford, MA, USA) water was used throughout the experiments.
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