Biomedicine & Pharmacotherapy 60 (2006) 182–185 http://france.elsevier.com/direct/BIOPHA/
Original article
Biotin supplementation reduces plasma triacylglycerol and VLDL in type 2 diabetic patients and in nondiabetic subjects with hypertriglyceridemia Cristina Revilla-Monsalve a, Iván Zendejas-Ruiz b, Sergio Islas-Andrade a, Armida Báez-Saldaña c, Miguel Angel Palomino-Garibay a, Pedro Martín Hernández-Quiróz d, Cristina Fernandez-Mejia b,* b
a Metabolic Diseases Unit, Mexican Institute of Social Security, Mexico Nutritional Genetics Unit, Biomedical Research Institute, Pediatric National Institute, National University of Mexico, Mexico c Department of Immunology, Biomedical Research Institute, National University of Mexico, Mexico d School of Medicine, National University of Mexico, Mexico
Received 16 February 2006; accepted 10 March 2006 Available online 31 March 2006
Abstract Biotin is a water-soluble vitamin that acts as a prosthetic group of carboxylases. Besides its role as carboxylase prosthetic group, biotin regulates gene expression and has a wide repertoire of effects on systemic processes. The vitamin regulates genes that are critical in the regulation of intermediary metabolism. Several studies have reported a relationship between biotin and blood lipids. In the present work we investigated the effect of biotin administration on the concentration of plasma lipids, as well as glucose and insulin in type 2 diabetic and nondiabetic subjects. Eighteen diabetic and 15 nondiabetic subjects aged 30–65 were randomized into two groups and received either 61.4 µmol/day of biotin or placebo for 28 days. Plasma samples obtained at baseline and after treatment were analyzed for total triglyceride, cholesterol, very low density lipoprotein (VLDL), glucose and insulin. We found that the vitamin significantly reduced (P = 0.005) plasma triacylglycerol and VLDL concentrations. Biotin produced the following changes (mean of absolute differences between 0 and 28 day treatment ± S.E.M.): a) triacylglycerol – 0.55 ± 0.2 in the diabetic group and –0.92 ± 0.36 in the nondiabetic group; b) VLDL: –0.11 ± 0.04 in the diabetic group and –0.18 ± 0.07 in the nondiabetic group. Biotin treatment had no significant effects on cholesterol, glucose and insulin in either the diabetic or nondiabetic subjects. We conclude that pharmacological doses of biotin decrease hypertriglyceridemia. The triglyceride-lowering effect of biotin suggests that biotin could be used in the treatment of hypertriglyceridemia. © 2006 Elsevier SAS. All rights reserved. Keywords: Biotin; Triacylglycerol; Type 2 diabetes
1. Introduction In the last few decades, an increasing number of vitaminmediated effects have been discovered at the level of gene expression [1] in addition to their well-known roles as substrates and cofactors [2]. The best recognized examples are the lipophilic vitamins A and D that serve as ligand precursors of the hormone nuclear receptors superfamily and thus affect functions such as morphogenesis, immunity, growth and epithelial cell differentiation [1]. The increasing knowledge of the molecular mechanisms of these vitamins has opened new perspectives that form a connection between nutritional signals and the development of new therapeutic agents [3,4]. Although little is known * Corresponding author. Av. del Iman #1, 4th floor, Mexico City, CP 04530, Mexico. E-mail address:
[email protected] (C. Fernandez-Mejia).
0753-3322/$ - see front matter © 2006 Elsevier SAS. All rights reserved. doi:10.1016/j.biopha.2006.03.005
about water-soluble vitamins as genetic modulators, there are increasing examples of their effects on gene expression [5–8]. Biotin is a water-soluble vitamin that acts as a prosthetic group of carboxylases. Unrelated to its role as carboxylase prosthetic group, biotin regulates gene expression. [7,8], and has a wide repertoire of effects on systemic processes such as development [9] immunity [10] and metabolism [11,12]. In rats, several studies have found that biotin regulates gene expression of critical genes involved in intermediary metabolism, as revealed by the mRNA levels of hepatic [13] and pancreatic [14,15] glucokinase (EC 2.7.1.1), insulin [15,16] and phosphoenolpyruvate carboxykinase (PEPCK; EC 4.1.1.32) [17]. Moreover, pharmacological doses of biotin appear to affect carbohydrate metabolism. Biotin administration ameliorates the diabetic state: hyperglycemia reduction was observed in both type 1 and type 2 diabetic patients, as well as in genetically diabetic KK mice and in OLETF rats, treated with biotin [18–21]. In hemodialysis
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patients, pharmacological doses of biotin improved their oral glucose tolerance tests [22]. Several investigations have reported a relationship between biotin and lipid metabolism [23–27]. Spontaneous symptoms of biotin deficiency were detected in rats genetically prone to development of elevated blood lipids [23]. In this rat strain, an inverse association was found between plasma lipids and biotin status [24]. A negative correlation between plasma biotin concentrations and blood lipids was also found in humans [25]. Furthermore, pharmacological doses of biotin modify plasma lipid concentrations: In healthy volunteers, 3.68 µmol/d of biotin supplementation affected plasma lipid concentrations [25]. In patients with hyperlipidemia, Dukusova and Krivoruchenko [26] found that a daily administration of 20.5 µmol biotin for 4 weeks decreased hypercholesterolemia. In a previous work [28] our findings suggested that the treatment with biotin (61.4 µmol/d) for 28 d decreased hypertriglyceridemia. There are no studies addressed to investigate the effect of biotin supplementation on hyperlipidemia in type 2 diabetic patients. In this work we compared the effect of biotin administration (61.4 µmol/day) on the lipid profile of type 2 diabetic patients and nondiabetic subjects with hypertriglyceridemia.
2.3. Blood collection and analytic methods
2. Materials and methods
3. Results
2.1. Subjects
The base line characteristics of the subjects participating in the study are provided in Table 1. The groups did not significantly differ in age, sex, or body mass index, triacylglycerol, VLDL, cholesterol or insulin concentrations. Fasting glucose concentrations were significantly different between the diabetic and the nondiabetic group. We investigated the effect of biotin administration on plasma triacylglycerol concentrations in either diabetic or nondiabetic hypertriglyceridemic subjects. Fig. 1 shows the effect of 28 d of biotin treatment on triacylglycerol concentrations in either diabetic or nondiabetic subjects. We found that biotin administration produced significant (P = 0.005) changes of –0.55 ± 0.2 in the diabetic group, and – 0.92 ± 0.36 in the nondiabetic group. No significant changes on triacylglycerol concentrations were observed in the placebo groups. We also investigated the effect of biotin on different metabolites and on insulin. Table 2 shows the mean of absolute differences between 0 and 28 d produced by the treatment with either biotin (61.4 µmol/d) or placebo on different metabolites in, either diabetic or nondiabetic, hypertriglyceridemic patients. Significant decreases (P = 0.005) in VLDL concentrations were observed after 28 d of biotin treatment (changes of – 0.11 ± 0.04 in the diabetic group, and –0.18 ± 0.07 in the nondiabetic group). Biotin treatment had no significant effects on fasting cholesterol, glucose and insulin, in either diabetic or nondiabetic subjects. Placebo administration did not significantly affect any of the measured metabolites in any group.
A total of 18 diabetic patients and 15 nondiabetic individuals took part in the study. The experimental protocol design used was reviewed and approved by the Research Committee of the Mexican Institute of Social Security (IMSS) in accordance with the Institute’s ethical standards in accordance with the Helsinky Declaration of 1975 as revised in 1983. Informed written consent was obtained from each subject previous to the start of the studies. All the studied subjects had plasma triacylglycerol concentrations of less than 5.33 mmol/l, and none had been treated with medication known to affect insulin sensitivity, glucose or lipid concentrations. Subjects had no prior history of renal, hepatic, immunologic or other endocrine disease. Active smokers, patients with a history of alcohol or drug abuse, or a systolic blood pressure over 160 mmHg were excluded from the study, as well as pregnant or lactating females. Subjects were instructed to follow their ordinary diet and daily activity level, and to not introduce any nutritional supplement for the duration of the experiment. Particularly, they were asked to avoid unusually lipid-rich meals two days before testing days. 2.2. Experimental design The protocol was a double blind, placebo-controlled study. This study consisted of a baseline phase followed by a 28 d treatment period during which subjects were randomly administered capsules containing either placebo (Avicel®, FMC, Mexico City, Mexico) or Avicel® plus 20.5 µmol (5 mg) biotin. Subjects were instructed to consume the test products 3 times/d before meals. On days 0 and 28 of treatment, the study subjects fasted overnight and were asked to arrive at the Metabolic Diseases Department at 7:00 h to draw blood samples.
Blood samples were collected in evacuated tubes (Beckton Dickson, Franklin Lakes, New Jersey, USA) for analysis of glucose, insulin, triacylglycerol and total cholesterol. Plasma glucose, triacylglycerol, VLDL and total cholesterol were measured with an automated analyzer (Kodak DT-60C; Kodak, Rochester, NY, USA). Plasma insulin concentrations were assessed with a commercially available radioimmunoassay kit (ICN, Costa Mesa, California, USA). All samples were analyzed in duplicate. 2.4. Statistical analysis Calculations were performed with the SPSS statistical package (version 11, SPSS Inc., Chicago, USA). The results are expressed as mean ± S.E.M. The Chi-square test was used to assess differences between sexes. The mean of absolute differences between 0 and 28 d were used to evaluate changes in plasma metabolites after treatment. Comparisons were evaluated by two-way ANOVA with interaction. A P value < 0.05 was considered statistically significant.
4. Discussion Results of the present study show that daily biotin supplementation with 61.4 µmol/d, during 28 d decreases triacylglycerol and VLDL concentrations in either diabetic or nondiabetic subjects, with hypertriglyceridemia. These results are in
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Table 1 Baseline characteristics of the participants in the study
Sex M/F Age BMI (kg/m2) Glucose* (mmol/l) Triacylglycerides (mmol/l) VLDL (mmol/l) Cholesterol (mmol/l) Insulin (nmol/l)
Placebo N = 8 3/5 49.0 ± 3.7 29.7 ± 1.36 11.90 ± 1.29 2.93 ± 0.31 0.59 ± 0.06 5.44 ± 0.31 139.2 ± 39.7
Diabetic Biotin N = 10 4/6 48.6 ± 3.49 29.7 ± 1.49 10.03 ± 0.95 3.34 ± 0.28 0.67 ± 0.06 5.59 ± 0.37 152.2 ± 19.9
Placebo N = 7 4/3 50.14 ± 5.05 28.4 ± 1.05 5.80 ± 0.18 2.73 ± 0.36 0.54 ± 0.07 5.87 ± 0.28 107.9 ± 20.6
Nondiabetic Biotin N = 8 5/3 46.9 ± 4.2 28.1 ± 0.97 5.72 ± 0.22 3.71 ± 0.44 0.74 ± 0.09 5.72 ± 0.47 135.6 ± 31.9
Placebo N = 7 0.33 ± 0.25 0.05 ± 0.07 –0.13 ± 0.11 0.18 ± 0.45 9.9 ± 19.0
Nondiabetic Biotin N = 8 –0.05 ± 0.14 –0.18 ± 0.07* –0.08 ± 0.10 –0.23 ± 0.33 –19.2 ± 23.0.
Mean ± S.E.M. * Significant difference between diabetic and nondiabetic * P < 0.005. Table 2 Mean changes values in plasma metabolites in the placebo and biotin-supplemented groups
BMI (kg/m2) VLDL (mmol/l) Glucose (mmol/l) Cholesterol (mmol/l) Insulin (nmol/l)
Placebo N = 8 0.21 ± 0.12 –0.01 ± 0.04 –0.26 ± 0.54 –0.16 ± 0.26 –11.5 ± 31.0
Diabetic Biotin N = 10 –0.37 ± 0.21 –0.11 ± 0.04 * –0.56 ± 0.62 –0.21 ± 0.18 –28.1 ± 20.8
Mean of absolute differences between 0 and 28 d of treatment ± S.E.M. * Significant difference between biotin and placebo treatments. * P < 0.005.
Fig. 1. Mean of absolute differences between 0 and 28 d (± S.E.M.) produced by the treatment with either biotin (61.4 µmol/d) or placebo on triacylglycerol concentrations in subjects, either diabetic or nondiabetic with hypertriglyceridemia. Left panel: Placebo administration; left column: diabetic; right column: nondiabetic. Right panel: Biotin treatment; left column: diabetic; right column: nondiabetic. Two-way ANOVA: (diabetic state and treatment). * Significant difference between biotin and placebo treatments (P = 0.005).
accordance with previous observations [25] showing that biotin treatment decreases triglyceridemia in healthy subjects and confirm and extend previous observations from our laboratory [28] that suggested that biotin treatment decreased hypertriglyceridemia in diabetic patients. We found that treatment with biotin did not modify plasma cholesterol. Studies by Dukusova and Krivoruchenko [26] have shown that the administration of biotin (2.05 µmol daily for 4 weeks) decreases blood cholesterol concentrations in patients with atherosclerosis and hyperlipidemia. The difference in response to biotin between above and present study could be related to the different cholesterol concentrations shown by the patients studied, namely 7.55 mmol/l in hypercholesterolemic patients in the Dukusova and Krivoruchenko study [26] versus 5.59 ± 0.37 mmol/l in our series. Indeed, several studies have found that pharmacological doses of biotin decrease blood cho-
lesterol concentrations in patients with exacerbated hyperlipidemia [25], but not in normolipemic subjects [27]. Hyperglycemia has been shown to be reduced by biotin treatment [18–21]. In Japanese type 2 diabetic patients, Maebashi et al. [19] found that biotin doses of 3.7 µmol/d for 1 month decreased fasting hyperglycemia. In the present study, as well as in a previous report [28], we found that biotin administration did not significantly decrease fasting glucose concentrations in diabetic patients. Possibly, genetic and nutritional differences between Japanese and Mexican diabetic account for the different responses to biotin. Studies in our laboratory are currently underway to outline the characteristics of biotin responsiveness in the diabetic subjects. We have shown that in vitro biotin increases insulin expression as well as glucose-induced insulin secretion [15]. The results obtained in the present study found that in vivo pharmacological doses of biotin do not significantly increase fasting insulin concentrations. Whether biotin treatment in vivo affects insulin expression and secretion, as observed in vitro, is an issue that is currently in progress in our laboratory. Little is known about the effects of pharmacological doses of biotin on the expression of lipid metabolism enzymes. In rats, Lewis et al. [29] found that rats consuming a diet containing 100 mg/kg of biotin during 21 days significantly reduced biotinylated ACC 2 abundance. ACC 2 inhibition has been shown to lead to a decrease in malonyl-CoA levels and the disinhibition of fatty acid oxidation [30]. Therefore, it may be possible that pharmacological doses of biotin increase fatty acid oxidation by decreasing ACC 2 activity. Current experiments in our laboratory are aimed at determining other molecular mechanisms by which biotin affects lipid metabolism Present findings suggest that biotin could be used in the treatment of hypertriglyceridemia. The notion that a vitamin can be used as a therapeutic agent in hypertriglyceridemia is not new. Indeed, another vitamin from the B complex, namely niacin, has
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been used since 1955 as a lipid-lowering drug [31]. Equally, extensive research on the molecular mechanisms of lipophilic vitamins A and D has opened new perspectives for the development of new retinoid- and calciferol-derived therapeutic agents [3,4], including promising candidates for diabetes treatment [32, 33]. It is only until recently that the molecular action of biotin and its effects on different metabolic functions has attracted attention. The important role of biotin in systemic functions is being reconsidered with the help of new technologies and with novel results regarding the molecular mechanisms of this vitamin. As with vitamins A and D, this research might lead to new perspectives in the development of therapeutic agents. 5. Conclusion Pharmacological doses of biotin decrease hypertriglyceridemia. The triglyceride-lowering effect of biotin suggests that biotin could be used in the treatment of hypertriglyceridemia. Acknowledgments We would like to thank Dr. Ignacio Camacho Arroyo and Dr. Silvestre Frenk for critical reading of this manuscript. We are indebted to José Luis García-Hernández for capsule preparation and to Alberto Rojas Ochoa for technical assistance. We also thank Isabel Pérez Montfort for correcting the English version of the manuscript. Supported by research grants from the Consejo Nacional de Ciencia y Tecnologia 34277M and 44266M and the Direccion General de Asuntos del Personal Academico IN201901. References [1]
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