Acute ethanol exposure inhibits renal folate transport, but repeated exposure upregulates folate transport proteins in rats and human cells.

Deficiency of folate in heavy-drinking alcoholic populations can occur partly because of an increased urinary folate excretion. Ethanol directly reduces the reabsorption of folate in the renal proximal tubule (PT) by acting on either of 2 folate transport proteins, the reduced folate carrier (RFC) and the folate receptor (FR). This study was designed to determine the effects of ethanol on the transport of folate by PT cells and to examine the effects of ethanol on RFC and the FR protein expression. Normal human PT (HPT) cells were cultured on membrane inserts to study intracellular transport of 5-methyltetrahydrofolate from the apical or basolateral direction in the presence of ethanol [11-109 mmol/L (50-500 mg/dL)]. The long-term effect of ethanol on the renal folate transport protein content was determined by western blot in treated HPT cells and in vivo in rats pair-fed control diets or ethanol-containing liquid diets. A 1-h treatment of HPT cells with ethanol (> or = 65 mmol/L) reduced the apically directed transport of folate by 20-25% without affecting the basolateral transport. A 5-d exposure of HPT cells to ethanol dose-dependently increased the content of both the FR and RFC proteins, with a greater effect on the RFC. Similarly, a 14-d exposure of rats to ethanol increased the in vivo expression of both the RFC and FR. These studies demonstrate that ethanol decreases the reabsorptive transport of folate by renal PT cells, which would increase urinary folate excretion. In contrast, subchronic exposure of PT cells, both in vivo and in vitro, to folate-depleting concentrations of ethanol leads to an upregulation of the 2 folate transport proteins. The increase in folate transporters partly counteracts the inhibitory effects of ethanol on folate transport activity, which explains the lower magnitude of ethanol's effect on transport with subchronic exposure compared with that with acute exposure.

Effects of HIV drug combinations on endothelin-1 and vascular cell proliferation.

Pulmonary arterial hypertension (PAH) is a progressive disease of the pulmonary vasculature involving endothelial and vascular smooth muscle cell (VSMC) proliferation, vasoconstriction, right ventricular hypertrophy, and eventually, right heart failure and death. PAH occurs 1000-fold more frequently in HIV patients than in the general population. Although conventional HIV therapy with nucleoside reverse transcriptase inhibitors (NRTIs) leads to regression of PAH, highly active antiretroviral therapy (HAART; two NRTI plus a protease inhibitor) increases the incidence of HIV-associated PAH as much as twofold. Although there are relatively few models for PAH, previous reports indicate the disease can be initiated by endothelial injury and release of the mitogen endothelin-1 (ET-1). ET-1, in turn, stimulates VSMC proliferation. To determine whether HAART induces endothelial injury and release of cytokines like ET-1, we treated human umbilical vein endothelial cells with micromolar amounts of AZT (3'-azido-3'-deoxythymidine), the protease inhibitor indinavir, or AZT plus indinavir, and measured cell viability, mitochondrial function, and ET-1 release. Both AZT and indinavir induced marked decreases in cellular oxygen uptake, as well as increases in ET-1 release. Although the drugs had no apparent effect on proliferation in VSMCs alone, in cocultures of VSMCs plus endothelial cells, the drugs increased proliferation of both endothelial cells and VSMCs. Finally, when cocultures of endothelial cells and VSMCs were treated with BQ-123 and BQ-788, selective antagonists for ET(A) and ET(B) receptors, respectively, drug-induced proliferation of both VSMCs and endothelial cells was attenuated. These data thus suggest that HIV drug cocktails may exacerbate preexisting HIV-associated PAH by inducing endothelial mitochondrial dysfunction, in turn stimulating the release of ET-1, and ultimately, vascular cell proliferation.

Effects of interleukin-1beta on food-maintained behavior in the mouse.

The present studies compared the effect of parenteral administration of the proinflammatory cytokine interleukin-1beta (IL-1beta) on food-seeking behavior under various conditions. IL-1beta (100 ng/mouse) decreased home cage consumption of sweetened milk to a greater extent in ad libitum fed mice than in mice that were food-restricted to maintain 85-90% of their free-feeding body weight. When operant responding for milk was maintained under a fixed-ratio 10 response (FR10) schedule of milk delivery, IL-1beta (30-300 ng/mouse) significantly decreased milk-maintained responding in mice fed ad libitum, but not in food-restricted mice. When food-restricted mice were trained under either an FR4 or FR32 response schedule of milk delivery, IL-1beta (100-300 ng/mouse) produced significant decreases in FR32, but not in FR4 responding. When responding was maintained under a progressive-ratio 10 response (PR10) schedule of milk delivery, IL-1beta (30-300 ng/mouse) dose-dependently decreased breaking points. These results indicate that the effects of IL-1beta on food-maintained behavior depend on both the level of motivation (as assessed by food restriction) and on the response cost for the milk (as assessed by ratio requirement). These findings suggest that motivational factors may be capable of attenuating some of the behavioral effects of these agents.