Oral spore-based probiotic supplementation was associated with reduced incidence of post-prandial dietary endotoxin, triglycerides, and disease risk biomarkers

Abstract

Aim: To determine if 30-d of oral spore-based probiotic supplementation could reduce dietary endotoxemia.

Methods: Apparently healthy men and women (n = 75) were screened for post-prandial dietary endotoxemia. Subjects whose serum endotoxin concentration increased by at least 5-fold from pre-meal levels at 5-h post-prandial were considered "responders" and were randomized to receive either placebo (rice flour) or a commercial spore-based probiotic supplement [Bacillus indicus (HU36), Bacillus subtilis (HU58), Bacillus coagulans, and Bacillus licheniformis, and Bacillus clausii] for 30-d. The dietary endotoxemia test was repeated at the conclusion of the supplementation period. Dietary endotoxin (LAL) and triglycerides (enzymatic) were measured using an automated chemistry analyzer. Serum disease risk biomarkers were measured using bead-based multiplex assays (Luminex and Milliplex) as secondary, exploratory measures.

Results: Data were statistically analyzed using repeated measures ANOVA and a P < 0.05. We found that spore-based probiotic supplementation was associated with a 42% reduction in endotoxin (12.9 ± 3.5 vs 6.1 ± 2.6, P = 0.011) and 24% reduction in triglyceride (212 ± 28 vs 138 ± 12, P = 0.004) in the post-prandial period Placebo subjects presented with a 36% increase in endotoxin (10.3 ± 3.4 vs 15.4 ± 4.1, P = 0.011) and 5% decrease in triglycerides (191 ± 24 vs 186 ± 28, P = 0.004) over the same post-prandial period. We also found that spore-based probiotic supplementation was associated with significant post-prandial reductions in IL-12p70 (24.3 ± 2.2 vs 21.5 ± 1.7, P = 0.017) and IL-1β (1.9 ± 0.2 vs 1.6 ± 0.1, P = 0.020). Compared to placebo post supplementation, probiotic subject had less ghrelin (6.8 ± 0.4 vs 8.3 ± 1.1, P = 0.017) compared to placebo subjects.

Conclusion: The key findings of the present study is that oral spore-based probiotic supplementation reduced symptoms indicative of "leaky gut syndrome".

Keywords: Cardiovascular disease; Chronic disease; High-fat meal challenge; Inflammatory cytokines; Leaky gut syndrome; Metabolic endotoxemia; Multiplex; Probiotics.

Figure 1

Represents the consort diagram for the study that indicates the number of participants that matriculated through the study. Subjects were carefully screened for exclusion/inclusion criteria and if qualified were enrolled in the study. Consistent with our preliminary data 2 out of every 6 subjects presented a dietary/metabolic endotoxin response following consumption of the high-fat meal. A total of 26 individuals were identified to have the “responder” phenotype and were randomized to participate in either the probiotic or placebo condition.

Figure 2

The change score for each variable compared to baseline was calculated and plotted as a heat map for probiotic vs placebo. Venous blood samples were collected prior to 3, and 5-h after consumption of a high-fat, high-calorie meal. Serum samples were analyzed using various accepted methods. Variables were divided into those that demonstrated a significant (upper panel) and those that did not (lower panel) have a significant probiotic effect. Responses were coded a lower (green to yellow) or higher (yellow to red) compared to baseline. An unchanged (yellow) response was also identified. TNF: Tumor necrosis factor; IL: Interleukin.

Figure 3

Serum endotoxin (A) and triglyceride (B) response to consumption of a commercially available high-fat, high-calorie pizza meal. ^aP < 0.05 indicates significantly less than placebo, less than pre-meal, less than and same time point at baseline. Venous blood samples were collected following an overnight fast and abstention from exercise. Serum samples were analyzed using an automated chemistry analyzer. Subjects consumed an oral probiotic supplement for 30-d and the experimental meal challenge was completed at baseline and following the 30-d supplementation period. Probiotic responses were compare to placebo. Panel C demonstrates individual variability in the dietary endotoxin response prior to and after the 30-d supplementation period. Red indicates a large dietary endotoxin response (> 5 fold increase from pre-meal) and green indicates a small dietary endotoxin response (< 1 fold increase from pre-meal).

Figure 4

Serum IL-12p70 (A), IL-1β (B), and ghrelin (C) response to consumption of a commercially available high-fat, high-calorie pizza meal. Venous blood samples were collected following an overnight fast and abstention from exercise. Serum samples were analyzed using an automated chemistry analyzer. Subjects consumed an oral probiotic supplement for 30-d and the experimental meal challenge was completed at baseline and following the 30-d supplementation period. Probiotic responses were compare to placebo. IL: Interleukin.

Figure 5

Serum IL-6 (A), IL-8 (B), and MCP-1 (C) response to consumption of a commercially available high-fat, high-calorie pizza meal. Venous blood samples were collected following an overnight fast and abstention from exercise. Serum samples were analyzed using an automated chemistry analyzer. Subjects consumed an oral probiotic supplement for 30-d and the experimental meal challenge was completed at baseline and following the 30-d supplementation period. Probiotic responses were compare to placebo. While effects did not reach statistical significance, trends are consistent with other variables that did significant change (Figures 2 and 3). IL: Interleukin.