Effects of human oral mucosal tissue, saliva, and oral microflora on intraoral metabolism and bioactivation of black raspberry anthocyanins.
Effects of human oral mucosal tissue, saliva, and oral microflora on intraoral metabolism and bioactivation of black raspberry anthocyanins.
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DOI：
10.1158/1940-6207.capr-11-0040
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发表时间：
2011-08
期刊：
影响因子：
--
通讯作者：
Liu Z
中科院分区：
文献类型：
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作者：
Mallery SR;Budendorf DE;Larsen MP;Pei P;Tong M;Holpuch AS;Larsen PE;Stoner GD;Fields HW;Chan KK;Ling Y;Liu Z
我们的口腔癌化学预防试验数据表明，冻干黑树莓（BRB）成分的局部保留和代谢的患者特异性差异影响治疗反应。随后的研究证实，花青素是BRB化学预防作用的关键因素。因此，进行功能测定、免疫印迹和免疫组织化学分析以评估BRB花青素相关代谢酶在人口腔组织中的水平和分布。对BRB漱口后收集的时程唾液样本进行LC-MS/MS分析，以评估局部药代动力学，并比较三种不同BRB漱口制剂提供持续口内花青素水平的能力。蛋白质谱表明，在所有15个口腔粘膜组织中存在的关键代谢酶的评价，而免疫组化证实这些酶分布在表面口腔上皮细胞和终端唾液导管。β-葡萄糖苷酶测定证实，全唾液和微生物区系减少的唾液可以使BRB花青素去糖基化，从而能够产生生物活性糖苷配基花青素。LC-MS/MS分析表明，在冲洗后，唾液中保留了亲本花青素及其功能性稳定代谢物原儿茶酸长达4小时。此外，冲洗后的唾液样本含有葡萄糖醛酸化的花青素缀合物，与细胞内摄取和BRB花青素转化为适合局部再循环的形式的第二阶段一致。我们的数据表明，与小肠相比，局部肠循环所需的水解酶、II相酶和外排转运酶在人口腔粘膜中存在并发挥作用。值得注意的是，患者间花青素生物活化和肠道再循环能力的差异将影响治疗，因为生物活化的化学预防剂在靶部位的保留将维持治疗有效性。
Our oral cancer chemoprevention trial data implied that patient-specific differences in local retention and metabolism of freeze-dried black raspberries' (BRB) components affected therapeutic responsiveness. Subsequent studies have confirmed that anthocyanins are key contributors to BRB's chemopreventive effects. Consequently, functional assays, immunoblotting and immunohistochemical analyses to evaluate levels and distribution of BRB anthocyanin-relevant metabolic enzymes in human oral tissues were performed. LC-MS/MS analyses of time course saliva samples collected following BRB rinses were conducted to assess local pharmacokinetics and compare the capacities of three different BRB rinse formulations to provide sustained intraoral levels of anthocyanins. Protein profiles demonstrated the presence of key metabolic enzymes in all 15 oral mucosal tissues evaluated while immunohistochemistry confirmed these enzymes were distributed within surface oral epithelia and terminal salivary ducts. β-glucosidase assays confirmed that whole and microflora-reduced saliva can deglycosylate BRB anthocyanins, enabling generation of the bioactive aglycone, cyanidin. LC-MS/MS analyses demonstrated retention of parent anthocyanins and their functional, stable metabolite, protocatechuic acid, in saliva for up to 4 hours after rinsing. Furthermore, post-rinse saliva samples contained glucuronidated anthocyanin conjugates, consistent with intracellular uptake and Phase II conversion of BRB anthocyanins into forms amenable to local recycling. Our data demonstrate that comparable to the small intestine, the requisite hydrolytic, Phase II and efflux transporting enzymes necessary for local enteric recycling are present and functional in human oral mucosa. Notably, inter-patient differences in anthocyanin bioactivation and capacities for enteric recycling would impact treatment as retention of bioactivated chemopreventives at the target site would sustain therapeutic effectiveness.