Student Scientific Association "Farmakon", Department of Biochemistry and Pharmacogenomics, Medical University of Warsaw, 1 Banacha Street, 02-097 Warsaw, Poland.
Polish Academy of Sciences Botanical Garden, Centre for Biological Diversity Conservation in Powsin, 2 Prawdziwka Street, 02-973 Warsaw, Poland.
Nutrients. 2022 May 30;14(11):2287. doi: 10.3390/nu14112287.
Targeting pancreatic lipase and α-amylase by digestion-derived fractions of ethanolic-aqueous (60%, /) extract from fruit (CM) in relation to the control and prevention of metabolic disorders, including diabetes, was the first purpose of the present study. Taking into consideration the significance of bio-accessibility of compounds, we attempted to identify metabolites of CM after gastrointestinal digestion in vitro, as well as their kinetic changes upon gut microbiota treatment. The digestion of extract was simulated with digestive enzymes in vitro and human gut microbiota ex vivo (1 h, 3 h, 6 h, 24 h), followed by chromatographic analysis using the UHPLC-DAD-MS method. The effect of fractions from gastrointestinal digestion in vitro on the activity of pancreatic lipase and α-amylase was studied with fluorescence-based assays. The gastric and intestinal fractions obtained after in vitro digestion of CM inhibited pancreatic lipase and α-amylase. Loganic acid as the main constituent of the extract was digested in the experimental conditions in contrast to cornuside. It was found in most analytes such as salivary, gastric, intestinal, and even colon (fecal slurry, FS) fractions. In all fractions, kaempferol hexoside and reduced forms of kaempferol, such as aromadendrin, and benzoic acid were assigned. The signals of tannins were detected in all fractions. Cornusiin A was tentatively assigned in the gastric fraction. The metabolites originating from kinetic analytes have been classified mainly as phenolic acids, hydrolyzable tannins, and flavonoids. Phenolic acids (protocatechuic acid, gallic acid), tannins (digalloylglucose, tri--galloyl--D-glucose), and flavonoids (aromadendrin, dihydroquercetin) were detected in the late phases of digestion in fecal slurry suspension. Cornuside was found in FS analyte after 3 h incubation. It was not detected in the samples after 6 and 24 h incubation with FS. In conclusion, cornuside, aromadendrin, and phenolic acids may be potentially bio-accessible compounds of CM. The presence of plants' secondary metabolites in the intestinal fractions allows us to indicate them as responsible for decreasing glucose and lipid absorption.
以消化衍生的乙醇-水溶液（60%，/）提取物的 fractions 为靶点，作用于胰腺脂肪酶和α-淀粉酶，这是本研究的首要目的。考虑到化合物生物可及性的重要性，我们试图鉴定 CM 在胃肠道消化后的代谢产物，以及它们在肠道微生物群处理下的动力学变化。采用体外消化酶和人体肠道微生物群（1 h、3 h、6 h、24 h）对提取物进行体外消化模拟，然后采用 UHPLC-DAD-MS 方法进行色谱分析。采用荧光法研究体外胃肠消化 fractions 对胰腺脂肪酶和α-淀粉酶活性的影响。CM 经体外消化后获得的胃和肠 fractions 抑制了胰腺脂肪酶和α-淀粉酶的活性。在实验条件下，与 cornuside 相反，作为提取物主要成分的 loganic acid 被消化。在唾液、胃、肠，甚至结肠（粪便浆，FS） fractions 中均发现了这种酸。在所有 fractions 中，均鉴定出山柰酚己糖苷和山柰酚的还原形式，如 aromadendrin 和苯甲酸。在所有 fractions 中均检测到单宁的信号。在胃 fractions 中，推测出 cornusiin A 的存在。源自动力学分析物的代谢物主要被分类为酚酸、可水解单宁和类黄酮。在粪便浆悬浮液的消化后期，检测到酚酸（原儿茶酸、没食子酸）、单宁（二没食子酰葡萄糖、三--没食子酰--D-葡萄糖）和类黄酮（aromadendrin、二氢槲皮素）。在 FS 分析物中，在 3 h 孵育后检测到 cornuside。在与 FS 孵育 6 和 24 h 后，未检测到该物质。总之，cornuside、aromadendrin 和酚酸可能是 CM 的潜在生物可利用化合物。肠道 fractions 中植物次生代谢物的存在使我们能够表明它们负责降低葡萄糖和脂质的吸收。
