Dietary Vitamin A Affects the Function of Incretin-Producing Enteroendocrine Cells in Male Mice Fed a High-Fat Diet.

BACKGROUND: Retinol-binding protein 2 (RBP2) is an intracellular carrier for vitamin A in the absorptive enterocytes. Mice lacking RBP2 (Rbp2-/-) display an unexpected phenotype of obesity, glucose intolerance, and elevated glucose-dependent insulinotropic polypeptide (GIP) levels. GIP and glucagon-like peptide 1 (GLP-1) are incretin hormones secreted by enteroendocrine cells (EECs). We recently demonstrated the presence of RBP2 and other retinoid-related proteins in EECs. OBJECTIVES: Given RBP2's role in intracellular retinoid trafficking, we aimed to evaluate whether dietary vitamin A affects incretin-secreting cell function and gene expression. METHODS: Male Rbp2-/- mice and sex- and age-matched controls (n = 6-9) were fed a high-fat diet (HFD) for 18 wk containing normal (VAN, 4000 IU/kg of diet) or low (VAL, 25% of normal) vitamin A concentrations. Body weight was recorded biweekly. Plasma GIP and GLP-1 levels were obtained fasting and 30 min after an oral fat gavage at week 16. Glucose tolerance tests were also performed. Mice were killed at week 18, and blood and tissue samples were obtained. RESULTS: Rbp2-/- mice displayed greater weight gain on the VAN compared with the VAL diet from week 7 of the intervention (P ≤ 0.01). Stimulated GIP levels were elevated in Rbp2-/- mice compared with their controls fed the VAN diet (P = 0.02), whereas their GIP response was lower when fed the VAL diet (P = 0.03). Although no differences in GLP-1 levels were observed in the VAN diet group, a lower GLP-1 response was seen in Rbp2-/- mice fed the VAL diet (P = 0.02). Changes in incretin gene expression and that of other genes associated with EEC lineage and function were consistent with these observations. Circulating and hepatic retinoid levels revealed no systemic vitamin A deficiency across dietary groups. CONCLUSIONS: Our data support a role for RBP2 and dietary vitamin A in incretin secretion and gene expression in mice fed a HFD.

Intestinal Enteroendocrine Cell Signaling: Retinol-binding Protein 2 and Retinoid Actions.

Retinol-binding protein 2-deficient (Rbp2-/-) mice are more prone to obesity, glucose intolerance, and hepatic steatosis than matched controls. Glucose-dependent insulinotropic polypeptide (GIP) blood levels are dysregulated in these mice. The present studies provide new insights into these observations. Single cell transcriptomic and immunohistochemical studies establish that RBP2 is highly expressed in enteroendocrine cells (EECs) that produce incretins, either GIP or glucagon-like peptide-1. EECs also express an enzyme needed for all-trans-retinoic acid (ATRA) synthesis, aldehyde dehydrogenase 1 family member A1, and retinoic acid receptor-alpha, which mediates ATRA-dependent transcription. Total and GIP-positive EECs are significantly lower in Rbp2-/- mice. The plasma transport protein for retinol, retinol-binding protein 4 (RBP4) is also expressed in EECs and is cosecreted with GIP upon stimulation. Collectively, our data support direct roles for RBP2 and ATRA in cellular processes that give rise to GIP-producing EECs and roles for RBP2 and RBP4 within EECs that facilitate hormone storage and secretion.

Retinol-binding protein 2 (RBP2): More than just dietary retinoid uptake.

Retinol-binding protein 2 (RBP2, also known as cellular retinol-binding protein 2 (CRBP2)) is a member of the fatty acid-binding protein family and has been extensively studied for its role in facilitating dietary vitamin A (retinol) uptake and metabolism within enterocytes of the small intestine. RBP2 is present in highest concentrations in the proximal small intestine where it constitutes approximately 0.1-0.5% of soluble protein. Recent reports have established that RBP2 binds monoacylglycerols (MAGs) with high affinity, including the canonical endocannabinoid 2-arachidonoylglycerol (2-AG). Crystallographic studies reveal that retinol, 2-AG, or other long-chain MAGs alternatively can bind in the retinol-binding pocket of RBP2. It also has been demonstrated recently that Rbp2-deficient mice are more susceptible to developing obesity and associated metabolic phenotypes when exposed to a high fat diet, or as they age when fed a conventional chow diet. When subjected to an oral fat challenge, the Rbp2-deficient mice release into the circulation significantly more, compared to littermate controls, of the intestinal hormone glucose-dependent insulinotropic polypeptide (GIP). These new findings regarding RBP2 structure and actions within the intestine are the focus of this review.

Retinol-binding protein 2 (RBP2): biology and pathobiology.

Retinol-binding protein 2 (RBP2; originally cellular retinol-binding protein, type II (CRBPII)) is a 16 kDa cytosolic protein that in the adult is localized predominantly to absorptive cells of the proximal small intestine. It is well established that RBP2 plays a central role in facilitating uptake of dietary retinoid, retinoid metabolism in enterocytes, and retinoid actions locally within the intestine. Studies of mice lacking Rbp2 establish that Rbp2 is not required in times of dietary retinoid-sufficiency. However, in times of dietary retinoid-insufficiency, the complete lack of Rbp2 gives rise to perinatal lethality owing to RBP2 absence in both placental (maternal) and neonatal tissues. Moreover, when maintained on a high-fat diet, Rbp2-knockout mice develop obesity, glucose intolerance and a fatty liver. Unexpectedly, recent investigations have demonstrated that RBP2 binds long-chain 2-monoacylglycerols (2-MAGs), including the canonical endocannabinoid 2-arachidonoylglycerol, with very high affinity, equivalent to that of retinol binding. Crystallographic studies establish that 2-MAGs bind to a site within RBP2 that fully overlaps with the retinol binding site. When challenged orally with fat, mucosal levels of 2-MAGs in Rbp2 null mice are significantly greater than those of matched controls establishing that RBP2 is a physiologically relevant MAG-binding protein. The rise in MAG levels is accompanied by elevations in circulating levels of the hormone glucose-dependent insulinotropic polypeptide (GIP). It is not understood how retinoid and/or MAG binding to RBP2 affects the functions of this protein, nor is it presently understood how these contribute to the metabolic and hormonal phenotypes observed for Rbp2-deficient mice.

Lipoprotein composition in patients with type 1 diabetes mellitus: Impact of lipases and adipokines.

OBJECTIVE: High cardiovascular mortality in patients with type 1 diabetes (T1DM) is widely recognized. Paradoxically, these patients have been shown to have elevated HDL-C and reduced apoB-containing lipoproteins. The purpose of this investigation was to further characterize the lipoprotein composition in T1DM and to assess the role that lipases and adipokines may play in these differences. METHODS: T1DM patients (89) attending the Diabetes Clinic at the University of Miami and 42 healthy controls were recruited. Clinical characteristics, lipoprotein composition (by ultracentrifugation and HPLC), leptin, and adiponectin were measured in the full cohort, while a subgroup had LPL and hepatic lipase measured. RESULTS: Subjects were predominately Caucasian and Hispanic. HgbA1c's were above goal while their mean duration of diabetes was >20 years. LPL was 2-fold elevated in diabetic women versus controls (+107%{p=0.001}) with no difference in men. Hepatic lipase was reduced 50% {p<0.001} in women but increased 50% {p=0.079} in men. Leptin was similar to controls in women but reduced in men (-60%{p<0.001}). Adiponectin was elevated in both genders (men: +55%{p=0.018}; women: +46%{p=0.007}). LDL-C was reduced in both diabetic men (-33%{p<0.001}) and women (-24%{p<0.001}) while HDL-C trended higher only in men (+13%{p=0.064}). Both total apoB (men: -31%{p<0.001}; women: -17%{p=0.016}) and triglycerides (men: -49%{p<0.001}; women: -31%{p=0.011}) were reduced in both genders while total apoA-I was increased in both (men: +31%{p<0.001}; women: +19%{p=0.008}). Both men and women had increases in LpA-I (+66%{p<0.001}; +40%{p=0.001}) which accounted for essentially the entire increase in HDL mass. VLDL lipids (men: -53→70%; women: -31→57%) were lower as was apoB (particle number) in men (-51{p<0.001}) with a similar trend in women (-35%{p=0.066}). Cholesterol esters in the particle core were depleted in both genders relative to both apoB (men: -41%; women: -37%) and triglycerides (men: -38%; women: -34%) (all{p<0.009}). There were similar differences in IDL. HDL-L lipids (except triglycerides) (men: +45→74%; women: +49→77%{p<0.006}), apoA-1 (men: +162%; women: +117%{p<0.001}), and apoA-II (men: +64%{p=0.008}; women: +55%{p=0.014}) were higher in T1DM patients. These differences produced dramatic increases in LpA-I (men: +221%; women +139%{p<0.001}) and total HDL-L mass (men: +85%; women: +78%{p<0.001}). ApoM (men: +190%; women: +149%{p<0.001}) was also dramatically increased. Conversely, HDL-D lipids were lower in both genders (-20%→50%) while apoA-I was not different in either. ApoA-II was lower only in the diabetic women (-25%{p=0.015}). LPL activity correlated primarily with IDL(-), LDL(-), HDL-L(+), and HDL-D(-) only in the women. HL correlated weakly with VLDL(+), LDL(+), HDL-L(-), and HDL-D(+) in women but had much stronger correlations with VLDL(-), IDL(-), and HDL-L(+). Adiponectin correlated with VLDL(-), IDL(-), LDL(-), HDL-L(+), and HDL-D(-) in women but only HDL-L(+) and HDL-D(-) in men. Leptin correlated with very few parameters in women but did correlate weakly with several HDL-L(-) and HDL-M(-) parameters. CONCLUSION: Lipoprotein composition and adipokine concentrations in both genders as well as lipase activities in the women would be expected to reduce the atherosclerotic risk in these patients with T1DM. These data suggest that there are functional lipoprotein abnormalities responsible for their CV risk that are not reflected in their plasma concentrations.

Elevated Lipoprotein Lipase Activity Does Not Account for the Association Between Adiponectin and HDL in Type 1 Diabetes.

CONTEXT: Increased high-density lipoprotein cholesterol (HDL-C) is common in type 1 diabetes (T1D) and is associated both with hyperadiponectinemia and with elevated lipoprotein lipase activity (LPL). Because adiponectin has been shown to increase LPL expression, elevated LPL may link the hyperadiponectinemia in T1D with increased HDL. OBJECTIVE: The purpose of this study was to determine whether LPL activity accounts for the association between adiponectin and HDL in T1D. DESIGN, PARTICIPANTS, AND SETTING: A cohort of 127 patients with T1D attending the Diabetes Clinic at the University of Miami and 103 healthy control subjects were recruited. MAIN OUTCOME MEASURE: HDL-C and adiponectin were measured in the full cohort and in a subgroup, HDL subfractions were obtained by ultracentrifugation, and LPL and hepatic lipase were measured in postheparin plasma. RESULTS: Total HDL-C and the lowest density HDL subfraction, apolipoprotein A-I, LPL activity, and adiponectin levels were higher in subjects with T1D than in control subjects (P < .05). Both adiponectin and LPL activity were directly associated with total HDL-C and its lowest density subfraction, but adiponectin and LPL were not correlated (P = 0.13). Adiponectin alone explained 11.6% and adiponectin plus LPL explained 23.8% of the HDL-C variance. In a multivariate model, adiponectin remained an independent predictor of HDL-C along with LPL and serum creatinine, explaining together 27% of HDL-C variance. CONCLUSIONS: Adiponectin was strongly associated with HDL-C in T1D, suggesting that hyperadiponectinemia is linked to the elevated HDL-C in this population. However, this relationship is independent of the association between LPL and HDL-C. Thus, elevated adiponectin and LPL activity are independently related to increased HDL-C in T1D.

Statins in the treatment of dyslipidemia in the presence of elevated liver aminotransferase levels: a therapeutic dilemma.

The beneficial role of statins in primary and secondary prevention of coronary heart disease has resulted in their frequent use in clinical practice. However, safety concerns, especially regarding hepatotoxicity, have driven multiple trials, which have demonstrated the low incidence of statin-related hepatic adverse effects. The most commonly reported hepatic adverse effect is the phenomenon known as transaminitis, in which liver enzyme levels are elevated in the absence of proven hepatotoxicity. This class effect is usually asymptomatic, reversible, and dose-related. However, the increasing incidence of chronic liver diseases, including nonalcoholic fatty liver disease and hepatitis C, has created a new challenge when initiating statin treatment in patients with high cardiovascular risk. These diseases result in abnormally high liver biochemistry values, discouraging statin use by clinicians, fostering treatment discontinuation, and leaving a large number of at-risk patients untreated. A PubMed/MEDLINE search of the literature regarding statin safety (January 1, 1994-December 31, 2008) was performed, using the following search terms: statin safety, statin-related hepatotoxicity, and chronic liver disease and statin use, as well as the specific names of different statins and different liver diseases. Relevant clinical trials, review articles, panel discussions, and guideline recommendations were selected. This review supports the use of statin treatment in patients with high cardiovascular risk whose elevated aminotransferase levels have no clinical relevance or are attributable to known stable chronic liver conditions. For each patient, the decision should be based on an individual assessment of risks and benefits.