Selective Influence of Hemp Fiber Ingestion on Post-Exercise Gut Permeability: A Metabolomics-Based Analysis

Abstract

Objectives: This study investigated the effects of 2-week ingestion of hemp fiber (high and low doses) versus placebo bars on gut permeability and plasma metabolite shifts during recovery from 2.25 h intensive cycling. Hemp hull powder is a rich source of two bioactive compounds, N-trans-caffeoyl tyramine (NCT) and N-trans-feruloyl tyramine (NFT), with potential gut health benefits. Methods: The study participants included 23 male and female cyclists. A three-arm randomized, placebo-controlled, double-blind, crossover design was used with two 2-week supplementation periods and 2-week washout periods. Supplement bars provided 20, 5, or 0 g/d of hemp hull powder. Participants engaged in an intensive 2.25 h cycling bout at the end of each of the three supplementation periods. Five blood samples were collected before and after supplementation (overnight fasted state), and at 0 h-, 1.5 h-, and 3 h-post-exercise. Five-hour urine samples were collected pre-supplementation and post-2.25 h cycling after ingesting a sugar solution containing 5 g of lactulose, 100 mg of ¹³C mannitol, and 1.9 g of mannitol in 450 mL of water. An increase in the post-exercise lactulose/¹³C mannitol ratio (L:¹³CM) was used as the primary indicator of altered gut permeability. Other outcome measures included muscle damage biomarkers (serum creatine kinase, myoglobin), serum cortisol, complete blood cell counts, and shifts in plasma metabolites using untargeted metabolomics. Results: No trial differences were found for L:¹³CM, cortisol, blood cell counts, and muscle damage biomarkers. Orthogonal partial least-squares discriminant analysis (OPLSDA) showed distinct trial differences when comparing high- and low-dose hemp fiber compared to placebo supplementation (R2Y = 0.987 and 0.995, respectively). Variable Importance in Projection (VIP) scores identified several relevant metabolites, including 3-hydroxy-4-methoxybenzoic acid (VIP = 1.9), serotonin (VIP = 1.5), 5-hydroxytryptophan (VIP = 1.4), and 4-methoxycinnamic acid (VIP = 1.4). Mummichog analysis showed significant effects of hemp fiber intake on multiple metabolic pathways, including alpha-linolenic acid, porphyrin, sphingolipid, arginine and proline, tryptophan, and primary bile acid metabolism. Conclusions: Hemp fiber intake during a 2-week supplementation period did not have a significant effect on post-exercise gut permeability in cyclists (2.25 h cycling bout) using urine sugar data. On the contrary, untargeted metabolomics showed that the combination of consuming nutrient-rich hemp fiber bars and exercising for 135 min increased levels of beneficial metabolites, including those derived from the gut in healthy cyclists.

Keywords: exercise; gut permeability; hemp fiber; metabolomics.

Figure 1

Study participant flow diagram. Thirty-eight participants were assessed for eligibility, and 25 were entered into the study, with 23 completing all aspects of the protocol.

Figure 2

Study design. This study employed a randomized, placebo-controlled, double-blind, crossover design with three 2-week supplementation periods and 2-week washout periods.

Figure 3

(a,b) OPLSDA analysis of high− and low−dose hemp study arms relative to placebo. Data represent metabolite changes from pre−supplementation time points across each of the four other time points (post−2 weeks supplementation, immediately post−2.25 h cycling, and 1.5 h and 3 h post-exercise.

Figure 4

Six metabolic pathways were significantly influenced by hemp fiber supplementation across all time points (interaction effects, p < 0.05). The color and size of each circle corresponds to its p-value and enrichment factor, respectively. The large red circles indicate the most significant pathways influenced by hemp fiber supplementation.

Figure 5

Four metabolic pathways were significantly influenced by hemp fiber supplementation across all time points (supplement main effects, p < 0.05). The color and size of each circle corresponds to its p-value and enrichment factor, respectively. The large red circles indicate the most significant pathways influenced by hemp fiber supplementation.