Stevia rebaudiana Bertoni, an American plant used as sweetener: Study of its effects on body mass control and glycemia reduction in Wistar male and female rats.

Stevia rebaudiana Bertoni water extracts have been used as a natural sweetener and customary medicine by the indigenous inhabitants of South America for several hundred years. This plant was sent to Europe in the 16th century and was described by Peter Jacob Esteve in Spain. Recently the food industry has started to employ S. rebaudiana as sweetener using its glycosides after purification. Advertisement claims that Stevia glycosides is good for controling body mass and reducing glycemia. This study's objective was to evaluate the effect of S. rebaudiana leaf extract on Wistar rats as animal model to prove its effectiveness on body mass control, glycemia reduction, and other biochemical parameters. Three groups were randomly formed with 24 males and 24 females: A blank group without any sweetener, a control group drinking water with 10% glucose, and the test group ingesting a 0.94% water extract of S. rebaudiana. Body mass measurements as well as food and drink consumption were daily performed. The experiment lasted 120 days after the specimens were weaned and got used to eating solid food. Euthanasia was done and blood serum was collected to evaluate the following biochemical parameters: Glucose, triglycerides, cholesterol, insulin, glucagon, leptin, ghrelin, and glucose-dependent insulinotropic peptide, GIP. Results indicated that only female rats had statistical differences in body mass gain. No relevant effects either positive or negative were found in the biochemical parameters measured. The crude extracts of S. rebaudiana did not show any relevant changes in biochemical and hormonal profiles, changes nor body mass with respect to the blank and control groups of young and healthy rats in the age range of infancy to youth. According to the results obtained, the therapeutic properties that have been associated to S. rebaudiana consumption especially for body mass control and glycemia reduction, did not occur in young and healthy male and female rats in equivalent age to infants, young children, and youths

Fig 3 -

A. Male Wistar rats daily food intake during 120 days after weaning consuming stevia infusion, drinking water sweetened with glucose (control), and plain water (blank). Each line represents the trend in the evaluated period where each point comprises the average food intake of each group and its CI at 95%, n = 8. B. Female Wistar rats daily food intake during 120 days after weaning consuming stevia infusion, drinking water sweetened with glucose (control), and plain water (blank). Each line represents the trend in the evaluated period where each point comprises the average food intake of each group and its CI at 95%, n = 8. C. Male Wistar rats statistical analyses of final cumulative food intake data 120 days after weaning consuming stevia infusion, drinking water sweetened with glucose (control), and plain water (blank). The following letters indicate significant differences p<0.05: a, b, c, d. Duncan test. D. Female Wistar rats statistical analyses of final cumulative food intake data 120 days after weaning consuming stevia infusion, drinking water sweetened with glucose (control), and plain water (blank). The following letters indicate significant differences p<0.05: a, b, c, d. Duncan test.</p

Methodology for preparing infusions and quantification of glucosides of interest (modified of [34]).

Methodology for preparing infusions and quantification of glucosides of interest (modified of [34]).</p

Fig 7 -

A. Male Wistar rats statistical analyses of serum insulin data 120 days after weaning consuming stevia infusion, drinking water sweetened with glucose (control), and plain water (blank). Its CI is at 95%. The following letters indicate significant differences p<0.05: a, b, c, d. Games-Howell Method n = 8. B. Female Wistar rats statistical analyses of serum insulin data 120 days after weaning consuming stevia infusion, drinking water sweetened with glucose (control), and plain water (blank). Its CI is at 95%. The following letters indicate significant differences p<0.05: a, b, c, d. Games-Howell Method n = 8. C. Male Wistar rats statistical analyses of serum glucagon data 120 days after weaning consuming stevia infusion, drinking water sweetened with glucose (control), and plain water (blank). Its CI is at 95%. The following letters indicate significant differences p<0.05: a, b, c, d. Games-Howell Method n = 8. D. Female Wistar rats statistical analyses of serum glucagon data 120 days after weaning consuming stevia infusion, drinking water sweetened with glucose (control), and plain water (blank). Its CI is at 95%. The following letters indicate significant differences p<0.05: a, b, c, d. Games-Howell Method n = 8. E. Male Wistar rats statistical analyses of serum leptin data 120 days after weaning consuming stevia infusion, drinking water sweetened with glucose (control), and plain water (blank). Its CI is at 95%. The following letters indicate significant differences p<0.05: a, b, c, d. Games-Howell Method n = 8. F. Female Wistar rats statistical analyses of serum leptin data 120 days after weaning consuming stevia infusion, drinking water sweetened with glucose (control), and plain water (blank). Its CI is at 95%. The following letters indicate significant differences p<0.05: a, b, c, d. Games-Howell Method n = 8. G. Male Wistar rats statistical analyses of serum ghrelin data 120 days after weaning consuming stevia infusion, drinking water sweetened with glucose (control), and plain water (blank). Its CI is at 95%. The following letters indicate significant differences p<0.05: a, b, c, d. Games-Howell Method n = 8. H. Female Wistar rats statistical analyses of serum ghrelin data 120 days after weaning consuming stevia infusion, drinking water sweetened with glucose (control), and plain water (blank). Its CI is at 95%. The following letters indicate significant differences p<0.05: a, b, c, d. Games-Howell Method n = 8. I. Male Wistar rats statistical analyses of serum glucose dependent insulinotropic peptide, GIP, data 120 days after weaning consuming stevia infusion, drinking water sweetened with glucose (control), and plain water (blank). Its CI is at 95%. The following letters indicate significant differences p<0.05: a, b, c, d. Games-Howell Method n = 8. J. Female Wistar rats statistical analyses of serum glucose dependent insulinotropic peptide, GIP, data 120 days after weaning consuming stevia infusion, drinking water sweetened with glucose (control), and plain water (blank). Its CI is at 95%. The following letters indicate significant differences p<0.05: a, b, c, d. Games-Howell Method n = 8.</p

S1 Raw data -

Stevia rebaudiana Bertoni water extracts have been used as a natural sweetener and customary medicine by the indigenous inhabitants of South America for several hundred years. This plant was sent to Europe in the 16th century and was described by Peter Jacob Esteve in Spain. Recently the food industry has started to employ S. rebaudiana as sweetener using its glycosides after purification. Advertisement claims that Stevia glycosides is good for controling body mass and reducing glycemia. This study’s objective was to evaluate the effect of S. rebaudiana leaf extract on Wistar rats as animal model to prove its effectiveness on body mass control, glycemia reduction, and other biochemical parameters. Three groups were randomly formed with 24 males and 24 females: A blank group without any sweetener, a control group drinking water with 10% glucose, and the test group ingesting a 0.94% water extract of S. rebaudiana. Body mass measurements as well as food and drink consumption were daily performed. The experiment lasted 120 days after the specimens were weaned and got used to eating solid food. Euthanasia was done and blood serum was collected to evaluate the following biochemical parameters: Glucose, triglycerides, cholesterol, insulin, glucagon, leptin, ghrelin, and glucose-dependent insulinotropic peptide, GIP. Results indicated that only female rats had statistical differences in body mass gain. No relevant effects either positive or negative were found in the biochemical parameters measured. The crude extracts of S. rebaudiana did not show any relevant changes in biochemical and hormonal profiles, changes nor body mass with respect to the blank and control groups of young and healthy rats in the age range of infancy to youth. According to the results obtained, the therapeutic properties that have been associated to S. rebaudiana consumption especially for body mass control and glycemia reduction, did not occur in young and healthy male and female rats in equivalent age to infants, young children, and youths.</div

Combined high-fat diet and sustained high sucrose consumption promotes NAFLD in a murine model

Background. The study of NAFLD in humans has several limitations. Using murine models helps to understand disease pathogenesis.Aim. Evaluate the impact of 4 different diets in the production of NAFLD with emphasis on a combined high-fat plus sustained high sucrose consumption.Material and methods. Eight week-old male Wistar rats were divided in four groups and fed for 90 days with the following diets: 1) Control chow diet (C); 2) High-fat cholesterol diet (HFC) + 5% sucrose in drinking water. 3) High-fat cornstarch diet (HFCO) + 5% sucrose in drinking water. 4) Chow diet + 20% sucrose in drinking water (HSD). Metabolic changes, leptin levels, liver histology, hepatic and plasma lipid composition, fasting plasma glucose and insulin and liver gene expression of FAS, SREBP-1 and PPAR-α were evaluated.Results. The HFC diet had the highest grade of steatosis (grade 2 of 3) and HSD showed also steatosis (grade 1). Liver weight TG and cholesterol concentrations in liver were greater in the HFC diet. There were no increased levels of iron in the liver. Rats in HFC gained significantly more weight (P < 0.001). All experimental groups showed fasting hyperglycemia. HFC had the highest glucose level (158.5 ± 7 mg/dL) (P < 0.005). The HSD and the HFCO diets developed also hyperglycemia. HSD had significantly higher fasting hyperinsulinemia. Serum leptin was higher in the HFC diet (p = 0.001). In conclusion, the HFC diet with combination of high fat and high sucrose is more effective in producing NAFLD compared with a high sucrose diet only

Fig 5 -

A. Male Wistar rats statistical analyses of theoretically calculated energy cumulative intake data 120 days after weaning consuming stevia infusion, drinking water sweetened with glucose (control), and plain water (blank). Its CI is at 95%. The following letters indicate significant differences p<0.05: a, b, c, d. Duncan test. B. Female Wistar rats statistical analyses of theoretically calculated energy cumulative intake data 120 days after weaning consuming stevia infusion, drinking water sweetened with glucose (control), and plain water (blank). Its CI is at 95%. The following letters indicate significant differences p<0.05: a, b, c, d. Duncan test.</p

Fig 6 -

A. Male Wistar rats statistical analyses of blood serum glucose data 120 days after weaning consuming stevia infusion, drinking water sweetened with glucose (control), and plain water (blank). Its CI is at 95%. The following letters indicate significant differences p<0.05: a, b, c, d. Games-Howell Method n = 8. B. Female Wistar rats statistical analyses of blood serum glucose data 120 days after weaning consuming stevia infusion, drinking water sweetened with glucose (control), and plain water (blank). Its CI is at 95%. The following letters indicate significant differences p<0.05: a, b, c, d. Games-Howell Method n = 8. C. Male Wistar rats statistical analyses of serum triglycerides data 120 days after weaning consuming stevia infusion, drinking water sweetened with glucose (control), and plain water (blank). Its CI is at 95%. The following letters indicate significant differences p<0.05: a, b, c, d. Games-Howell Method n = 8. D. Female Wistar rats statistical analyses of serum triglycerides data 120 days after weaning consuming stevia infusion, drinking water sweetened with glucose (control), and plain water (blank). Its CI is at 95%. The following letters indicate significant differences p<0.05: a, b, c, d. Games-Howell Method n = 8. E. Male Wistar rats statistical analyses of serum cholesterol data 120 days after weaning consuming stevia infusion, drinking water sweetened with glucose (control), and plain water (blank). Its CI is at 95%. The following letters indicate significant differences p<0.05: a, b, c, d. Games-Howell Method n = 8. F. Female Wistar rats statistical analyses of serum cholesterol data 120 days after weaning consuming stevia infusion, drinking water sweetened with glucose (control), and plain water (blank). Its CI is at 95%. The following letters indicate significant differences p<0.05: a, b, c, d. Games-Howell Method n = 8.</p

Fig 4 -

A. Male Wistar rats daily water intake during 120 days after weaning consuming stevia infusion, drinking water sweetened with glucose (control), and plain water (blank). Each line represents the trend in the evaluated period where each point comprises the average water intake of each group and its CI at 95%, n = 8. B. Female Wistar rats daily water intake during 120 days after weaning consuming stevia infusion, drinking water sweetened with glucose (control), and plain water (blank). Each line represents the trend in the evaluated period where each point comprises the average water intake of each group and its CI at 95%, n = 8. C. Male Wistar rats statistical analyses of final cumulative water intake data 120 days after weaning consuming stevia infusion, drinking water sweetened with glucose (control), and plain water (blank). The following letters indicate significant differences p<0.05: a, b, c, d. Duncan test. D. Female Wistar rats statistical analyses of final cumulative water intake data 120 days after weaning consuming stevia infusion, drinking water sweetened with glucose (control), and plain water (blank). The following letters indicate significant differences p<0.05: a, b, c, d. Duncan test.</p

Fig 2 -

A. Male Wistar rats final cumulative body mass gain 120 days after weaning consuming a water infusion of S. rebaudiana dried leaves, drinking water with glucose (control), and plain water (blank). Each line represents the trend in the evaluated period where each point comprises the average body mass gain of the group and its CI at 95%, n = 8. B. Female Wistar rats final cumulative body mass gain 120 days after weaning consuming a water infusion of S. rebaudiana dried leaves, drinking water with glucose (control), and plain water (blank). Each line represents the trend in the evaluated period where each point comprises the average body mass gain of the group and its CI at 95%, n = 8. C. Male Wistar rats statistical analyses of final cumulative body mass gain data 120 days after weaning consuming a water infusion of S. rebaudiana dried leaves, drinking water with glucose (control), and plain water (blank). The following letters indicate significant differences pS. rebaudiana dried leaves, drinking water with glucose (control), and plain water (blank). The following letters indicate significant differences p<0.05: a, b, c, d. Duncan test.</p