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      Is Open Access

      An imaging approach for determining the mechanism of enhancement of intestinal absorption of an L-theanine supplement

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          Abstract

          Background & objective

          Theanine (L-glutamylethylamide) contained in green tea is a functional food component that has been attracting attention due to its relaxation effect. It was shown that the ingredients added to the theanine formulations increased the absorption of theanine. If this mechanism can be elucidated, it would be possible to contribute to development of evidence-based formulations. In this study, we investigated the effect of ingredients in the formulations on the absorption of theanine in detail.

          Main methods

          After oral administration of a mixture of theanine and additional components to Wistar rats the plasma concentration was determined by an HPLC and the pharmacokinetic parameters were calculated. In addition, a new system for evaluating intestinal blood flow was developed since the involvement of intestinal blood flow was considered as a factor that increased absorption of theanine.

          Key findings

          Plasma concentration of theanine increased significantly in the combined use group with eight ingredients containing piperine as compared with theanine only group. Piperine would increase theanine absorption by increased blood flow, not an inhibition of metabolism. We succeeded to develop a visual and quantitative system to evaluate the effect of these ingredients directly including piperine on the intestinal blood flow using indocyanine green while maintaining physiological conditions.

          Significance

          Increased intestinal blood flow by these ingredients including piperine enhanced the absorption of theanine. Other mechanisms may also be considered as the mechanism by which theanine absorption is increased in addition to increased blood flow.

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          Most cited references 36

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          Protein measurement with the Folin phenol reagent.

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            Black pepper and its pungent principle-piperine: a review of diverse physiological effects.

             K Srinivasan (2006)
            Black pepper (Piper nigrum) is one of the most widely used among spices. It is valued for its distinct biting quality attributed to the alkaloid, piperine. Black pepper is used not only in human dietaries but also for a variety of other purposes such as medicinal, as a preservative, and in perfumery. Many physiological effects of black pepper, its extracts, or its major active principle, piperine, have been reported in recent decades. Dietary piperine, by favorably stimulating the digestive enzymes of pancreas, enhances the digestive capacity and significantly reduces the gastrointestinal food transit time. Piperine has been demonstrated in in vitro studies to protect against oxidative damage by inhibiting or quenching free radicals and reactive oxygen species. Black pepper or piperine treatment has also been evidenced to lower lipid peroxidation in vivo and beneficially influence cellular thiol status, antioxidant molecules and antioxidant enzymes in a number of experimental situations of oxidative stress. The most far-reaching attribute of piperine has been its inhibitory influence on enzymatic drug biotransforming reactions in the liver. It strongly inhibits hepatic and intestinal aryl hydrocarbon hydroxylase and UDP-glucuronyl transferase. Piperine has been documented to enhance the bioavailability of a number of therapeutic drugs as well as phytochemicals by this very property. Piperine's bioavailability enhancing property is also partly attributed to increased absorption as a result of its effect on the ultrastructure of intestinal brush border. Although initially there were a few controversial reports regarding its safety as a food additive, such evidence has been questionable, and later studies have established the safety of black pepper or its active principle, piperine, in several animal studies. Piperine, while it is non-genotoxic, has in fact been found to possess anti-mutagenic and anti-tumor influences.
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              Indocyanine green: observations on its physical properties, plasma decay, and hepatic extraction.

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                Author and article information

                Contributors
                Role: ConceptualizationRole: Data curationRole: Formal analysisRole: Funding acquisitionRole: InvestigationRole: MethodologyRole: VisualizationRole: Writing – original draft
                Role: ConceptualizationRole: Data curationRole: Formal analysisRole: InvestigationRole: MethodologyRole: Validation
                Role: Formal analysisRole: InvestigationRole: MethodologyRole: ResourcesRole: SoftwareRole: Visualization
                Role: Visualization
                Role: ConceptualizationRole: Funding acquisitionRole: Project administrationRole: ResourcesRole: SoftwareRole: SupervisionRole: VisualizationRole: Writing – review & editing
                Role: Editor
                Journal
                PLoS One
                PLoS One
                plos
                PLoS ONE
                Public Library of Science (San Francisco, CA USA )
                1932-6203
                11 June 2021
                2021
                : 16
                : 6
                Affiliations
                [1 ] Faculty of Pharmaceutical Sciences, Hokkaido University, Sapporo, Hokkaido, Japan
                [2 ] Global Station for Biosurfaces and Drug Discovery, Global Institution for Collaborative Research and Education (GI-CoRE), Hokkaido University, Sapporo, Hokkaido, Japan
                [3 ] Department of Pharmacy, Hokkaido University Hospital, Sapporo, Hokkaido, Japan
                Shenzhen Bay Laboratory, CHINA
                Author notes

                Competing Interests: The authors have declared that no competing interests exist.

                Article
                PONE-D-20-38266
                10.1371/journal.pone.0253066
                8195392
                34115818
                © 2021 Sato et al

                This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

                Page count
                Figures: 4, Tables: 1, Pages: 16
                Product
                Funding
                Funded by: The Japan Society for the Promotion of Sciences
                Award ID: 16K00842, 20K07153
                Award Recipient :
                Funded by: Grants-in-Aid for Regional R&D Proposal-Based Program from Northern Advancement Center for Science & Technology of Hokkaido
                Award Recipient :
                Funded by: Hokkaido University, Global Facility Center (GFC), Pharma Science Open Unit (PSOU), funded by the Ministry of Education, Culture, Sports, Science and Technology (MEXT) under “Support Program for Implementation of New Equipment Sharing System
                Award Recipient :
                Funded by: The Platform Project for Supporting in Drug Discovery and Life Science Research (Platform for Drug Discovery, Informatics, and Structural Life Science) from the MEXT and Japan Agency for Medical Research and Development (AMED)
                Award Recipient :
                This work was supported in part by Grants-in-Aid for Regional R&D Proposal-Based Program from Northern Advancement Center for Science & Technology of Hokkaido and Grants-in-Aid for Scientific Research (C) (Grant number 16K00842, 20K07153) from the Japan Society for the Promotion of Science (JSPS). This research was partly supported by Hokkaido University, Global Facility Center (GFC), Pharma Science Open Unit (PSOU), funded by the Ministry of Education, Culture, Sports, Science and Technology (MEXT) under “Support Program for Implementation of New Equipment Sharing System.” This research was also partially supported by the Platform Project for Supporting in Drug Discovery and Life Science Research (Platform for Drug Discovery, Informatics, and Structural Life Science) from the MEXT and Japan Agency for Medical Research and Development (AMED).
                Categories
                Research Article
                Biology and Life Sciences
                Anatomy
                Digestive System
                Gastrointestinal Tract
                Medicine and Health Sciences
                Anatomy
                Digestive System
                Gastrointestinal Tract
                Biology and Life Sciences
                Anatomy
                Body Fluids
                Blood
                Blood Flow
                Medicine and Health Sciences
                Anatomy
                Body Fluids
                Blood
                Blood Flow
                Biology and Life Sciences
                Physiology
                Body Fluids
                Blood
                Blood Flow
                Research and Analysis Methods
                Imaging Techniques
                Fluorescence Imaging
                Medicine and Health Sciences
                Pharmacology
                Pharmacokinetics
                Drug Absorption
                Biology and Life Sciences
                Anatomy
                Body Fluids
                Blood
                Blood Plasma
                Medicine and Health Sciences
                Anatomy
                Body Fluids
                Blood
                Blood Plasma
                Biology and Life Sciences
                Physiology
                Body Fluids
                Blood
                Blood Plasma
                Medicine and Health Sciences
                Pharmacology
                Routes of Administration
                Oral Administration
                Research and Analysis Methods
                Biological Cultures
                Cell Lines
                Caco-2 Cells
                Research and Analysis Methods
                Bioassays and Physiological Analysis
                Biochemical Analysis
                Bioelectrochemical Analysis
                Amperometry
                Custom metadata
                All relevant data are within the paper and its Supporting information files.

                Uncategorized

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