Abstract
To (1) investigate the efficacy of multiple doses of an orally delivered probiotic bacteria Lactobacillus paracasei (LP) modified to express angiotensin (1–7) (LP-A) in altering physiologic parameters relevant to the gut-brain axis in older rats and to (2) compare this strategy with subcutaneous delivery of synthetic Ang(1–7) peptide on circulating Ang(1–7) concentrations and these gut-brain axis parameters. Male 24-month-old F344BN rats received oral gavage of LP-A, or subcutaneous injection of Ang(1–7) for 0×, 1×, 3×, or 7×/week over 4 weeks. Circulating RAS analytes, inflammatory cytokines, and tryptophan and its downstream metabolites were measured by ELISA, electrochemiluminescence, and LC-MS respectively. Microbiome taxonomic analysis of fecal samples was performed via 16S-based PCR. Inflammatory and tryptophan-related mRNA expression was measured in colon and pre-frontal cortex. All dosing regimens of LP-A induced beneficial changes in fecal microbiome including overall microbiota community structure and α-diversity, while the 3×/week also significantly increased expression of the anti-inflammatory species Akkermansia muciniphila. The 3×/week also increased serum serotonin and the neuroprotective analyte 2-picolinic acid. In the colon, LP-A increased quinolinate phosphoribosyltransferase expression (1×/week) and increased kynurenine aminotransferase II (1× and 3×/week) mRNA expression. LP-A also significantly reduced neuro-inflammatory gene expression in the pre-frontal cortex (3×/week: COX2, IL-1β, and TNFα; 7×/week: COX2 and IL-1β). Subcutaneous delivery of Ang(1–7) increased circulating Ang(1–7) and reduced angiotensin II, but most gut-brain parameters were unchanged in response. Oral—but not subcutaneous—Ang(1–7) altered physiologic parameters related to gut-brain axis, with the most effects observed in 3×/week oral dosing regimen in older rats.
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Acknowledgments
We would like to thank Brian Bouverat and Liana Juarez for assistance with specific technical aspects of the projects. We would also like to thank Richard Franklin, MD, PhD of Constant Therapeutics for the donation of TXA 127 (Ang[1-7]) for the study involving subcutaneous administration of Ang(1-7).
Funding
This work was funded by a grant from the National Institutes of Health/National Institute on Aging (R01AG054538). The work was also supported by the following: The UAB Nathan Shock Center (P30AG50886), Microbiome Resource (supported by NIH grants P30CA013148 and UL1TR001417), Metabolism Core Laboratory (supported by P30DK56336, P30DK079626, and U4TR001368), and O’Brien Center Bioanalytical Core (supported by P30DK079337).
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Buford, T.W., Sun, Y., Roberts, L.M. et al. Angiotensin (1–7) delivered orally via probiotic, but not subcutaneously, benefits the gut-brain axis in older rats. GeroScience 42, 1307–1321 (2020). https://doi.org/10.1007/s11357-020-00196-y
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DOI: https://doi.org/10.1007/s11357-020-00196-y