Improvement of Lipoprotein Profile and Metabolic Endotoxemia by a Lifestyle Intervention That Modifies the Gut Microbiota in Subjects With Metabolic Syndrome

Abstract

Background Metabolic syndrome (MetS) is a serious health problem over the world; thus, the aim of the present work was to develop a lifestyle intervention to decrease the dysbiosis of gut microbiota and reduce the biochemical abnormalities of MetS. Methods and Results The prevalence of MetS was evaluated in 1065 subjects of Mexico City, Mexico, and the gut microbiota in a subsample. Subjects with MetS were selected for a pragmatic study based on a lifestyle intervention with a low-saturated-fat diet, reduced-energy intake, with functional foods and physical activity, and a second group was selected for a randomized control-placebo study to assess the gut microbiota after the dietary intervention. Prevalence of MetS was 53%, and the higher the body mass index, the higher the gut microbiota dysbiosis. The higher the Homeostatic Model Assessment for Insulin Resistance, the lower the high-density lipoprotein cholesterol concentration. The pragmatic study revealed that after 15 days on a low-saturated-fat diet, there was a 24% reduction in serum triglycerides; and after a 75-day lifestyle intervention, MetS was reduced by 44.8%, with a reduction in low-density lipoprotein cholesterol, small low-density lipoprotein particles, glucose intolerance, lipopolysaccharide, and branched-chain amino acid. The randomized control-placebo study showed that after the lifestyle intervention, there was a decrease in the dysbiosis of the gut microbiota associated with a reduction in the Prevotella/ Bacteroides ratio and an increase in the abundance of Akkermansia muciniphila and Faecalibacterium prausnitzii. Conclusions A lifestyle intervention significantly decreased MetS components, small low-density lipoprotein particle concentration, gut microbiota dysbiosis, and metabolic endotoxemia, reducing the risk of atherosclerosis. Clinical Trial Registration URL: https://www.clinicaltrials.gov. Unique identifier: NCT03611140.

Keywords: functional foods; lipoprotein; metabolic endotoxemia; metabolic syndrome; microbiota.

Figures

Figure 1

Physiological/biochemical parameters according to BMI categories in 1065 participants. A, Fat mass (%) and lean mass (%). B, Serum glucose and insulin. C, Relative frequency of subjects with Homeostatic Model Assessment for Insulin Resistance (HOMA‐IR) ≥2.5. D, Serum triglyceride concentration. E, Serum high‐density lipoprotein cholesterol (HDL‐C) concentration in women and men. F, Serum leptin. G, Serum CRP (C‐reactive protein). To compare the variables among body mass index categories, 1‐way ANOVA, followed by Tukey's post hoc test, was performed. Different letters indicate significant differences among groups: a>b>c>d (P<0.05). NW indicates normal weight; OW, overweight; OCI, class I obesity; OCII, class II obesity; OCIII, class III obesity.

Figure 2

Biochemical parameters according to body mass index (BMI) categories and presence of metabolic syndrome (MetS) in 1032 participants. A, Percentage of participants with and without MetS according to the BMI category: overweight (OW), class I obesity (OCI), class II obesity (OCII), and class III obesity (OCIII). B, Percentage of participants with and without MetS, according to BMI categories and sex. C, Systolic and diastolic blood pressure, according to BMI categories. Circles in blue represent without MetS, and triangles in red represent with MetS. D, Serum glucose and insulin concentration, according to BMI categories. E, Homeostatic Model Assessment for Insulin Resistance (HOMA‐IR), according to BMI categories; the red bars represent with MetS, and the blue bars represent without MetS. F, Serum triglyceride concentration, according to BMI categories. G, High‐density lipoprotein (HDL) cholesterol concentration, according to BMI categories and sex. The differences in biochemical/physiological parameters between participants with and without MetS were determined using an unpaired Student t test.

Figure 3

Metabolic syndrome (MetS) modifies the gut microbiota in Mexican adults. A, Principal coordinate analysis (PCoA) of normal weight (NW) subjects and subjects with MetS or class III obesity (OCIII)+MetS on the basis of the weighted UniFrac distances. The red squares represent samples of healthy subjects, blue triangles represent subjects with MetS, and orange circles represent subjects with OCIII+MetS. B, The α diversity by Shannon index in NW subjects indicates higher diversity in healthy subjects than subjects with OCIII+MetS. C and D, Taxonomic summary of the gut microbiota of NW subjects and subjects with MetS and OCIII+MetS at the phylum (C) and at the genus (D) levels. E, Serum lipopolysaccharide (LPS) in healthy subjects, subjects with MetS, and subjects with OCIII+MetS. F, Discriminative taxa at the species level in NW subjects and subjects with MetS and OCIII+MetS were determined using a linear discriminant analysis (LDA) effect size. The green bar chart represents the species that were more abundant in MetS and OCIII+MetS subjects, and the red bar chart represents the healthy subjects.

Figure 4

Relationship between specific genera and metabolic endotoxemia and body mass index categories and carbohydrate intake. A, Prevotella/Bacteroides ratio in normal weight (NW) subjects and subjects with metabolic syndrome (MetS) and class III obesity (OCIII)+MetS; statistical analysis was performed using 1‐way ANOVA, followed by Fisher's Least Significant Difference (LSD) post hoc test. Different letter indicates significant differences among groups: a>b>c (P<0.05). B, Correlation between serum lipopolysaccharide and Prevotella copri. The statistical analysis was performed using the correlation of Spearman. C, Association between Prevotella/Bacteroides ratio and percentage carbohydrates or percentage protein consumed in the diet.

Figure 5

Effect of a lifestyle intervention with functional foods and energy reduction (−500 kcal) for 75 days on clinical and biochemical characteristics in 146 patients with metabolic syndrome. In the first stage, participants were instructed to consume a reduced‐energy diet for 15 days. This period is indicated by a vertical dotted line. During the second stage of the study, participants consumed the dietary intervention and functional foods in addition to the reduced‐energy diet for 60 days. A, Waist circumference in all patients and separated by sex. B, Body mass index (BMI) in all patients and separated by sex. C, Serum triglycerides. D, Systolic and diastolic blood pressure. E, Serum glucose. F, Serum high‐density lipoprotein (HDL) cholesterol separated by sex. Statistical analysis was performed using 1‐way ANOVA, followed by Bonferroni post hoc test. Logarithmic transformation was performed before the statistical analysis. Different letters indicate significant differences among groups: a>b>c>d (P<0.05).

Figure 6

Effect of a lifestyle intervention with functional foods and energy reduction for 75 days on body composition and biochemical parameters in 146 patients with metabolic syndrome (MetS). In the first stage, participants were instructed to consume a reduced‐energy diet for 15 days. During the second stage of the study, participants consumed the functional foods in addition to the reduced‐energy diet for 60 days. A, Serum leptin concentration, according to body mass index (BMI) category (overweight [OW], class I obesity [OCI], and class II obesity [OCII]) and sex. B, Percentage of fat mass in all patients and separated by sex. C, Homeostatic Model Assessment for Insulin Resistance (HOMA‐IR) index. D, Percentage of glycosylated hemoglobin (HbA1c) in all patients and according to BMI category. E, Glucose serum concentration after an oral glucose tolerance test (OGTT) at 0 and 75 days after the dietary strategy in all patients and area under the curve (AUC), according to the BMI category. F, Insulin concentration after an OGTT at 0 and 75 days after the dietary strategy in all patients and AUC for insulin, according to BMI category. G, Serum lipopolysaccharide. H, Serum branched‐chain amino acid (BCAA), according to BMI category (n=111). Statistical analysis was performed using 1‐way ANOVA, followed by Bonferroni post hoc test. Logarithmic transformation was performed before the statistical analysis. Different letters indicate significant differences among groups: a>b>c>d (P<0.05). Comparisons between 2 groups were analyzed by a paired Student t test. Significant differences are shown. *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001.

Figure 7

Effect of lifestyle intervention with functional foods and energy reduction for 75 days on lipoprotein profile in 146 patients with metabolic syndrome. A, Serum total cholesterol concentration. B, Nuclear magnetic resonance–calculated lipids: plasma triglycerides (TG), very‐low‐density lipoprotein (VLDL), chylomicron triglycerides (CTGs), and high‐density lipoprotein (HDL) cholesterol. C, Plasma VLDL and chylomicron particle concentration: VLDL and chylomicron particles (VLDLCPs), large VLDL and chylomicron particles (VLCPs), medium VLDL particles (VMPs) and small VLDL particles (VSPs). D, Plasma low‐density lipoprotein concentration (LDL): total LDL particles, intermediate‐density lipoprotein (IDL) particles, large LDL particles (LARGE), and small LDL particles (SMALL). E, Plasma HDL particle concentration: total HDL particle (TOTAL), large HDL particles (LARGE), medium HDL particles (MEDIUM), small HDL particles (SMALL). F, Mean particle size of VLDL, LDL, and HDL. Statistical analysis was performed by a paired Student t test. Significant differences are shown. *P<0.05.

Figure 8

Effect of a lifestyle intervention on changes in anthropometric and biochemical parameters, according to the presence of specific polymorphisms in patients with metabolic syndrome. A, High‐density lipoprotein (HDL) cholesterol. B, Triglycerides. C, Very‐low‐density lipoprotein (VLDL) and chylomicron triglyceride concentration after dietary strategy, according to the presence of ADIPOQ genotype. D, Serum intermediate‐density lipoprotein (IDLP) after dietary strategy, according to the presence of FTO genotype. E, Serum LDL cholesterol after dietary strategy, according to the presence of ABCA1 genotypes. F, Weight. G, Fat mass. H, Body mass index (BMI) after dietary strategy, according to the presence of GFOD2 genotypes. I, Serum LDL particles after dietary strategy, according to presence of APOE isoforms. J, Serum area under the curve (AUC) of glucose after dietary strategy, according to presence of TCF7L2 genotypes.

Figure 9

Effect of a lifestyle intervention with functional foods (FFs) or placebo (P) on gut microbiota in patients with metabolic syndrome (MetS). A, The α diversity by Shannon index, according to the body mass index category after dietary intervention (DI) with placebo or FFs. Normal weight (placebo=9, FFs=11), MetS (placebo=17, FFs=18), class III obesity (OCIII)–MetS (placebo=10, FFs=13). B, Taxonomic summary of gut microbiota at the genus level. C, Prevotella/Bacteroides ratio after the dietary strategy with placebo or FFs. Discriminative taxa at the species level in normal weight (NW; D), MetS (E), and OCIII+MetS (F) subjects using a linear discriminant analysis (LDA) effect size. The green bar chart represents the species that were more abundant in subjects consuming the DI+placebo, and the red bar chart represents the subjects consuming the DI+FFs. Comparisons between 2 groups were analyzed by an unpaired Student t test. Significant differences are shown. *P<0.05, **P<0.01, ****P<0.0001.