Use of a lactobacillus rhamnosus strain for reducing weight gain and/or insulin resistance

Abstract

Provided are the use of Lactobacillus rhamnosus strain CNCM I-3690 for reducing diet-induced weight gain and/or diet-induced insulin resistance, and treating disorders resulting therefrom, such as overweight, obesity and obesity-related disorders in a subject.

Claims

1. A method of reducing diet-induced weight gain and/or diet-induced insulin resistance in a subject, comprising administering to a subject in need thereof a fermented food product comprising at least 10.sup.6 CFU/g of a Lactobacillus rhamnosus strain deposited at the Collection Nationale de Cultures de Micro-organismes (CNCM) with accession number I-3690.

2. The method of claim 1, wherein the method is for the treatment, or alleviation of a condition resulting from diet-induced weight gain and/or diet-induced insulin resistance, and said condition is selected from the group consisting of being overweight and obesity.

3. The method of claim 1, wherein said fermented food product is a fermented dairy product.

Description

FIGURE LEGENDS

(1) FIG. 1: Weight gain (A), fasting blood glucose (B), fasting insulin (C), HOMA-IR (D), OGTT (E) and areas under the curve (AUC) of OGTT (F) for four groups: NC, HFD, HFD+CNCM I-3690, HFD+B. lactis B420 (Danisco). Data are shown as means S.E.M. **p<0.01, *p<0.05 when compared to HFD group, and ##p<0.01, #p<0.05 when compared to NC group by One Way-ANOVA followed by Tukey post hoc test in SPSS. HOMA-IR is calculated according to the following formula: fasting blood glucose (mmol/L)fasting insulin (mU/L)/22.5.

(2) FIG. 2: Food intake of four groups each week. Data are shown as means of two cages of mice, so no statistical analysis was performed.

(3) FIG. 3: Cumulative food intake of four groups in each month of the animal trial. Data are shown as means of two cages of mice, so no statistical analysis was performed.

(4) FIG. 4: Cumulative food intake of four groups during 12 weeks. Data are shown as means of two cages of mice, so no statistical analysis was performed.

EXAMPLE 1

Improvement of High Fat Diet-Induced Obesity and Insulin Resistance By L. Rhamnosus CNCM 1-3690.

(5) Materials and Methods

(6) C57BL/6J mice (male, at age 12 weeks) were divided into 3 groups (8 mice per group) under different treatments as follows:

(7) Group A: high fat diet, containing 34.9% fat, 5.24 kcal/g, from Research Diets, Inc., New Brunswick, N.J. (HFD);

(8) Group B: high fat diet, plus probiotic strain L. rhamnosus CNCM 1-3690, at 10.sup.8 CFU/mouse/day (HFD+CNCM 1-3690);

(9) Group C: high fat diet, plus probiotic strain Bifidobacterium lactis B420 (Danisco), at 10.sup.8 CFU/mouse/day (HFD+B. lactis B420), previously reported to reduce adverse effects on metabolism associated with high-fat diet (AMAR et al., 2011, cited above), as a comparison;

(10) Group D; Normal chow, containing 4.3% fat, 3.85 kcal/g, from Research Diets, Inc., New Brunswick, N.J. (NC).

(11) L. rhamnosus CNCM 1-3690 or Bifidobacterium lactis B420 suspension were prepared before the animal trial, stored at 80 C. and thawed 1 hour before they were administered to each mouse by oral feeding.

(12) Animal treatments lasted for 12 weeks, during which the body weight of each mouse and food intake of every cage of mice were measured twice a week. Fresh stool and urine samples were collected once a month by using a metabolic cage and immediately stored at 80 C. for subsequent analysis.

(13) The amount of the probiotic strains in the feces of mice at 2nd, 6th and 11th weeks during the probiotic administration was quantified by reverse transcription (RT)-qPCR, and the results confirmed that they could survive in the intestine.

(14) At the end of the trial, after 5 h of food deprivation, blood was collected from the orbital plexus, and serum was isolated by centrifugation at 3000 rpm at 4 C. for 15 min. All animals were sacrificed by cervical dislocation. Epididymal fat pads, liver and jejunum were excised, weighed, and immediately kept in RNALater (Ambion) after sacrifice.

(15) Oral glucose tolerance tests (OGTT) were performed before the sacrifice of animals. After 5 h of food deprivation, 2.0 g/kg body weight glucose was administered orally to the mice. Blood samples were taken from the tail to measure blood glucose levels before and 15, 30, 60, and 120 min after glucose administration by using an ACCU-Check glucose meter (Roche Diagnostics, Canada). The blood glucose level before glucose administration is regarded as fasting blood glucose (FBG) level. Fasting insulin (FINS) level was determined by ELISA assay (Mercodia, Sweden). HOMA-IR was calculated according to the following formula: fasting blood glucose (mmol/L)fasting insulin (mU/L)/22.5.

(16) Results

(17) The results are shown in FIG. 1: A: Body weight gain; B: Fasting blood glucose levels (FBG); C: Fasting insulin levels (FINS); D: homeostasis assessment of insulin resistance (HOMA-IR) index; E: Curve of Oral Glucose Tolerance Test (OGTT), F: Areas under the curve (AUC) of OGTT. Data are shown as means S.E.M. **p<0.01, *p<0.05 when compared to HFD group, and ##p<0.01, #p<0.05 when compared to NC group by One Way-ANOVA followed by Tukey post hoc test in SPSS.

(18) Compared with NC-fed mice, the HFD group showed higher weight gain (FIG. 1A), elevated levels of fasting blood glucose (FIG. 1B), of fasting insulin (FIG. 1C), and of HOMA-IR (FIG. 1D), as well as decreased glucose tolerance (FIG. 1E, F). The supplement of two probiotic strains to HFD feeding significantly decreased the body weight gain (FIG. 1A). Although there was no significant difference in Fasting Blood glucode (FBG) and Fasting insulin (FINS) levels between HFD+probiotics groups and HFD group both L. rhamnosus CNCM 1-3690 or Bifidobacterium lactis B420 reduced the HOMA-IR index, and this reduction was significant in the case of CNCM 1-3690 (FIG. 1D). The two strains also significantly decreased glucose intolerance (FIG. 1E, F), indicating that they could improve insulin resistance.

(19) The average energy intake per mouse per day (FIG. 2) was calculated for each of the twelve weeks of the trial. During all the trial, the energy intake of NC group was the lowest, and the energy intake of HFD+probiotic groups was almost the same with that of the HFD group except for the 7.sup.th week. Moreover, cumulative energy intake of four groups of animal during 3 months (FIG. 3) and cumulative energy intake of four groups of animal during 12 weeks (FIG. 4) were calculated. This indicates that the body weight reduction observed for the probiotic treated groups cannot be attributed to a reduction of the energy intake.

(20) These results show that the two probiotic strains significantly improved the obesity and insulin resistance induced by HFD, and that the improvement provided by L. rhamnosus strain CNCM I-3690 is at least comparable to that provided by Bifidobacterium lactis B420.