Quercetin to enhance the bioavailability and activity of green tea polyphenols

槲皮素增强绿茶多酚的生物利用度和活性

• 批准号: 8641331
• 负责人: Susanne Margarete Henning
• 金额: $ 7.47万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-04-01 至 2016-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8641331
• 关键词: ABCC1 gene; Adverse effects; Animals; Apple; Berry; Biological; Biological Availability; Blood; Broccoli - dietary; Carrier Proteins; Catechol O-Methyltransferase; Cell Culture Techniques; Cell Line; Chemical Structure; Chemicals; Chemoprevention; Chemopreventive Agent; Clinical Trials; Combined Modality Therapy; Consumption; CpG Islands; Cytosine; DNA; DNA Methylation; DNA Methyltransferase; DNA Modification Methylases; Data; Detection; Diagnosis; Dose; Double-Blind Method; Enterohepatic Circulation; Enzyme Gene; Epigallocatechin Gallate; Erythrocytes; Evaluation; Excretory function; Flavonoids; Foundations; Future; Gene Expression; Gene Proteins; Genes; Green tea; Hepatotoxicity; High Pressure Liquid Chromatography; Hour; Human; Hypermethylation; Induction of Apoptosis; Intake; Intervention; Intervention Studies; LNCaP; MSH3 gene; Malignant neoplasm of prostate; Marketing; Metabolism; Methylation; Methyltransferase; Multi-Drug Resistance; Multidrug Resistance Associated Protein 1; Mus; Onions; Participant; Phase; Placebo Control; Placebos; Plasma; Prevention; Promoter Regions; Prostate; Prostatectomy; Prostatic Neoplasms; Public Health; Published Comment; Quercetin; Randomized; Research Design; SCID Mice; Schedule; Staging; Tea; Testing; Time; Tissues; Toxic effect; Tumor Tissue; Tumor Volume; Urine; Western Blotting; Xenograft procedure; absorption; arm; base; capsule; carcinogenesis; cell growth; drinking; enzyme activity; epicatechin; epicatechin gallate; epigallocatechin-(4-8,2-O-7)epicatechin; gallocatechol; genome-wide; inhibitor/antagonist; men; polyphenol; prevent; prostate cancer model; prostatitis; protein expression; public health relevance; subcutaneous; sulfation; tissue processing; tumor; tumor growth; uptake

DESCRIPTION (provided by applicant): Based on the reviewers' comments and new findings we revised the study design, dose, timing, tissue processing and statistical evaluation. The objective of the proposed human intervention study remains to determine whether a combined administration of quercetin and green tea (GT) will increase the bioavailability and decrease the metabolism of GT polyphenols (GTPs) such as epigallocatechin gallate (EGCG). Factors limiting the biological activity of GTPs are the reduced bioavailability and extensive metabolism converting EGCG into methylated metabolites with decreased activity. In our previous human intervention study in men drinking 6 cups of green tea daily prior to prostatectomy we demonstrated that GTPs are present in human prostate tissue. However 50 percent of EGCG was found in methylated form. Quercetin, a flavonoid found in onions, apples, broccoli, berries and teas is a natural inhibitor of catechol-O-methyltransferase (COMT) and multidrug resistance transport proteins (MRP1 and MRP2). Our cell culture studies demonstrated that the combined treatment with EGCG and quercetin led to a 4 to 10-fold increase in intracellular concentration of EGCG and significant decrease in EGCG methylation, depending on the cell line used. We demonstrated in mouse prostate cancer xenograft studies that the administration of the combination of GT with quercetin significantly decreased tumor volume, increased EGCG tissue concentration and decreased the methylation of EGCG significantly compared to treatment with GT or quercetin alone. The combined administration also significantly decreased the protein expression of methyltransferases and MRP1. We therefore propose a placebo- controlled, double-blinded, randomized, parallel two arm human pilot intervention study to determine whether the consumption of GT extract and quercetin will increase the bioavailability of GTPs in blood and prostate tissue and decrease the methylation of EGCG. We will administer two capsules of GT extract combined with one capsule of quercetin (N=25) or with one placebo capsule (N=15) twice daily for 3 weeks to men scheduled for prostatectomy. After 10 participants will complete the intervention, liver toxicity will be evaluated. In the absence of lier toxicity the quercetin dose will be doubled (800 mg daily) (N=15). GTPs, quercetin and methylated metabolites will be analyzed in blood and prostate tissue using high performance liquid chromatography (HPLC) with coularray electrochemical detection (ECD). To evaluate the mechanism involved, COMT and DNA methyltransferase enzyme activities and gene and protein expression as well as MRP1 gene and protein expression and global DNA methylation will be determined in tumor and normal prostate tissue and COMT activity in erythrocytes using HPLC-ECD, real time qPCR and Western blot analyses. The results will lay the foundation for future clinical trials to evaluate the interaction of quercetin and GT in chemoprevention of early stages of prostate cancer.

描述（由申请人提供）：根据审稿人的意见和新发现，我们修改了研究设计、剂量、时间、组织处理和统计评估。拟议的人类干预研究的目的仍然是确定槲皮素和绿茶 (GT) 的联合给药是否会增加 GT 多酚 (GTP) 如表没食子儿茶素没食子酸酯 (EGCG) 的生物利用度并降低 GT 多酚 (GTP) 的代谢。限制 GTP 生物活性的因素是生物利用度降低以及将 EGCG 转化为活性降低的甲基化代谢物的广泛代谢。在我们之前的人类干预研究中，男性在前列腺切除术前每天喝 6 杯绿茶，我们证明了 GTP 存在于人类前列腺组织中。然而，50% 的 EGCG 是以甲基化形式存在的。槲皮素是一种存在于洋葱、苹果、西兰花、浆果和茶叶中的黄酮类化合物，是儿茶酚-O-甲基转移酶 (COMT) 和多药耐药性转运蛋白（MRP1 和 MRP2）的天然抑制剂。我们的细胞培养研究表明，EGCG 和槲皮素联合处理可使细胞内 EGCG 浓度增加 4 至 10 倍，并且 EGCG 甲基化显着降低，具体取决于所使用的细胞系。我们在小鼠前列腺癌异种移植研究中证明，与单独使用 GT 或槲皮素治疗相比，GT 与槲皮素联合用药可显着减小肿瘤体积，增加 EGCG 组织浓度，并显着降低 EGCG 甲基化。联合用药还显着降低了甲基转移酶和 MRP1 的蛋白表达。因此，我们提出了一项安慰剂对照、双盲、随机、平行的两臂人类试点干预研究，以确定摄入 GT 提取物和槲皮素是否会增加血液和前列腺组织中 GTP 的生物利用度并降低 EGCG 的甲基化。我们将为计划进行前列腺切除术的男性每天两次服用两粒 GT 提取物胶囊与一粒槲皮素胶囊 (N=25) 或一粒安慰剂胶囊 (N=15) 联合服用，持续 3 周。 10 名参与者完成干预后，将评估肝脏毒性。在没有其他毒性的情况下，槲皮素剂量将加倍（每天 800 毫克）（N=15）。将使用高效液相色谱 (HPLC) 和 Coularray 电化学检测 (ECD) 来分析血液和前列腺组织中的 GTP、槲皮素和甲基化代谢物。为了评估所涉及的机制，将使用 HPLC-ECD、实时 qPCR 和蛋白质印迹分析测定肿瘤和正常前列腺组织中的 COMT 和 DNA 甲基转移酶活性、基因和蛋白质表达以及 MRP1 基因和蛋白质表达和整体 DNA 甲基化以及红细胞中的 COMT 活性。该结果将为未来评价槲皮素和GT在前列腺癌早期化学预防中相互作用的临床试验奠定基础。