Thérèse Rouleau

Photooxidation of Parenteral Multivitamins Induces Hepatic Steatosis in a Neonatal Guinea Pig Model of Intravenous Nutrition

Photooxidation of multivitamin solutions results in the generation of peroxides. Because peroxides are associated with hepatic steatosis and fibrosis as well as cholestasis, we questioned whether multivitamins are implicated in hepatic complications of parenteral nutrition. Guinea pig pups were assigned to groups receiving intravenously either total parenteral nutrition, photo-protected or not, or a control solution (5% dextrose + 0.45% NaCl) supplemented with either a) multivitamins; b) photo-protected multivitamins; c) multivitamins without riboflavin; or d) peroxides (H2O2, tert-butylhydroperoxide). After 4 d, liver was sampled for histology and isoprostane-F2α levels, a marker of radical attack. Multivitamins as well as total parenteral nutrition were associated with steatosis (scored 0–4), the severity of which was reduced (p < 0.05) by photo protection. Although H2O2 is the major peroxide contaminating multivitamins, it did not induce steatosis scores different than the controls. Compared with controls, hepatic isoprostane-F2α content increased in animals infused with H2O2 (p < 0.05), but not in those infused with Multi-12 pediatric multivitamins or total parenteral nutrition. Results suggest that peroxides and/or free radicals are not mediators of the induction of steatosis observed with infusion of photo-exposed multivitamins, as there was no correspondence between histologic findings and hepatic levels of isoprostanes. It is suspected that a component of the multivitamin solution becomes hepato-toxic after photo-exposure, as indicated by the protective effect observed when withdrawing riboflavin. Photo-oxidation of multivitamins might be the common link between reports involving amino acids, lipids, and light exposure in the ethiology of hepatic complications of parenteral nutrition.

Determinants of oxidant stress in extremely low birth weight premature infants

Early in life, premature neonates are at risk of oxidant stress. They often require total parenteral nutrition (TPN), which is, however, contaminated with oxidation products. Coadministration of parenteral multivitamins (MVP) with a lipid emulsion (LIP) prevents lipid peroxidation. We hypothesized that LIP+MVP induces a lower oxidant load compared to preparations in which MVP is administered with an amino acid solution (AA+MVP). The aim of this study was to compare markers of oxidant stress in premature neonates receiving LIP+MVP, either exposed to or protected from light, or AA+MVP. Antioxidant vitamins, the redox potential of glutathione, isoprostane, and dityrosine were measured in urine or blood sampled on days 7 and 10 from babies requiring low (<0.25) vs high (≥0.25) fractional inspired O(2). Oxygen supplementation induced a more oxidized redox potential and increased dityrosine with AA+MVP only. Adding MVP in the lipid rather than the amino acid moiety of TPN protects against the oxidant stress associated with O(2) supplementation. Photoprotection added no benefit. Blood transfusions were found to produce a pronounced oxidant load masking the beneficial effect of LIP+MVP. The impact of these findings relates to a strong association between a more oxidized redox potential and later bronchopulmonary dysplasia, a clinical marker of oxidant stress.

Peroxide-like oxidant response in lungs of newborn guinea pigs following the parenteral infusion of a multivitamin preparation

The multivitamin solution is a major component responsible for the photo-induced generation of peroxides in parenteral nutrition. The lung is a target of oxidant injury; however, the specific role of infused peroxides is unknown. The aim of this study was to determine if parenteral multivitamins induce in the lung an oxidant challenge similar to that of peroxides. Newborn guinea pigs were infused with dextrose plus relevant concentrations of H(2)O(2) (0,250,500 microM) or multivitamins (0,1%), as well as parenteral nutrition supplemented with multivitamins (0,1%). After 4 days, total glutathione, glutathione-related enzymes, and oxidant-sensitive eicosanoids were measured in the lungs. Peroxides as well as multivitamins led to a significant decrease in glutathione and the activity of glutathione synthase, indicating that infused peroxides were not entirely transformed into free radicals, which would have stimulated glutathione synthesis. The multivitamin solution induced a response in oxidant-sensitive eicosanoids similar to the response to peroxides, suggesting an oxidant stress that was not alleviated by the antiradical properties of its components. The effects on prostaglandins occurred independently from the stimulation in glutathione levels induced by parenteral nutrition. The multivitamin solution carries an oxidant load and causes effects similar to those of peroxides in the lungs of newborn guinea pigs.

Parenteral multivitamin supplementation induces both oxidant and antioxidant responses in the liver of newborn guinea pigs.

Background The multivitamin solution is a major component of photo-induced generation of peroxides in parenteral nutrition. The aim of this study was to determine whether the parenteral multivitamin preparation induces in the liver a peroxide-induced oxidant challenge or an antioxidant protection associated with the antiradical components of the solution. Methods Newborn guinea pigs were infused with dextrose supplemented with peroxides (250 &mgr;mol/L H2O2 or 350 &mgr;mol/L tert-butylhydroperoxide) or with a multivitamin preparation (MVP, 1% vol/vol). After 4 days, total glutathione and a free radical–sensitive eicosanoid marker (prostaglandin I2 [PGI2]/total prostaglandins) were measured in livers. Results There was a significant decrease in the PGI2/total prostaglandin ratio (mean ± SEM ) [dextrose: 0.068 ± 0.007 vs. (dextrose + H2O2: 0.048 ± 0.001, dextrose + TBH: 0.043 ± 0.001)] and glutathione concentrations decreased [dextrose: 55 ± 7 vs. (dextrose + H2O2: 37 ± 7, dextrose + TBH: 18 ± 7 nmol/mg protein)] after infusion of peroxides. Despite the peroxide load in the multivitamin solution, it did not alter the measured variables as prostanoid ratio remained at control concentrations (dextrose: 0.066 ± 0.008 vs. dextrose + MVP: 0.065 ± 0.006), as did glutathione levels (dextrose: 52 ± 6 vs. dextrose + MVP: 45 ± 7 nmol/mg prot). Conclusion In the liver of guinea pig pups, infused peroxides cause oxidation of membrane-derived prostanoids. The decrease in glutathione in response to administration of peroxides suggests consumption rather than a response to a free radical attack. Despite the oxidant load associated with peroxides generated in MVP, the multivitamin preparation protected membranes as the prostanoid ratio, and glutathione levels remained at control levels.

Influence of lung oxidant and antioxidant status on alveolarization : Role of light-exposed total parenteral nutrition

Parenteral multivitamins (MVP) are linked to the generation of peroxides, which cause oxidant injury in lungs associated with alveolar remodelling linked to lung disease of prematurity. This study was to investigate the relationship between alveolar development and lung oxidant-antioxidant status as modulated by the mode of administration of multivitamins with total parenteral nutrition (TPN). Four groups of guinea pig pups received parenteral nutrition differing by 1) mode of MVP admixture: with amino acid solution (AA-MVP) or lipid emulsion (LIP-MVP); 2) light exposure: TPN exposed (LE) or shielded from light (LP). After 2 or 4 days of TPN, vitamins C and E, 8-isoprostaneF2alpha and alveolarization index were determined in lungs and GSSG/GSH in lungs and blood. Exposure to light and the mode of MVP admixture did not influence vitamin E and isoprostane levels. Blood glutathione redox potential was more oxidized in LE and LIP-MVP groups after 4-day infusions, whereas lung redox potential was more reduced in LE groups. LP and LIP-MVP had a beneficial effect, with higher number of alveoli. Globally, results indicate that in this model, alveolarization and modifications in lung redox potential are two independent events induced by light exposed TPN.

Inflammatory Response in Preterm Infants Is Induced Early in Life by Oxygen and Modulated by Total Parenteral Nutrition

The i.v. lipid emulsion (LIP) is a source of oxidants, which may stimulate inflammation. Coadministration of parenteral multivitamins (MVP) with LIP prevents lipid peroxidation in light-exposed total parenteral nutrition (TPN). We hypothesized that this modality of TPN administration affects systemic inflammation, which may be modulated by exposure to oxygen. Premature infants were allocated to three TPN regimens: control regimen — MVP coadministered with amino acid/dextrose exposed to ambient light, LIP provided separately (n = 9) — LIP+MVP light exposed (LE): MVP coadministered with light-exposed LIP (n = 9) — LIP+MVP light protected (LP): MVP coadministered with light-protected LIP (n = 8). In LE and LP, amino acid/dextrose was provided separately. On reaching full TPN, infants were sampled for IL-6 and IL-8 in plasma and the redox potential of glutathione in whole blood (E, mV). Data were compared (ANOVA) in infants exposed to low (<0.25) versus high (≥0.25) FiO2. Patients (mean ± SD: birth weight 797 ± 172 g; GA 26 ± 1 wk) had similar clinical characteristics in TPN groups. Cytokine levels correlated positively (p < 0.01) with FiO2 and E. High FiO2 stimulated an increase (p < 0.01) in cytokines in control regimen, whereas these markers remained unaffected by oxygen in the LE and LP groups. The choice of a TPN admixture may have important consequences on the systemic inflammatory response triggered by an oxidant stress.

Photoprotection prevents TPN-induced lung procollagen mRNA in newborn guinea pigs.

BACKGROUND Photo-exposed intravenous multivitamin solutions (MVP) carry a peroxide load. Peroxidation induces gene expression of procollagen. We hypothesized that photo exposure of the MVP solution might promote pulmonary fibrosis. The aim of the study was to assess the potential for MVP to increase procollagen mRNA. METHODS Three day old guinea pigs were assigned to the following intravenous regimens, either: Control (C): 5% dextrose + 0.45% NaCl; C + 200 or 500 microM H(2)O(2); C + 500 microM H(2)O(2) + 10 microM GSSG; [C + 1% MVP +/- [amino acids + lipids]] +/- photoprotected. After 4 d, levels of pulmonary alpha1(I) procollagen mRNA and glutathione were determined. Results were compared by ANOVA. RESULTS Photoprotection of MVP or TPN prevents light induction of procollagen mRNA. The effect of MVP + light was associated with a peroxide load coupled with a low glutathione level. This was also observed with the 500 microM H(2)O(2) group. The addition of GSSG prevented the increase of procollagen mRNA caused by H(2)O(2). CONCLUSION An oxidant stress caused by the infusion of peroxides in an organism with a weak antiperoxide capacity induces the transcription of the gene encoding for procollagen alpha1(I). The results confirm the antiperoxide activity of lung glutathione. Parenteral nutrition could be a clinical condition favoring the initiation of lung fibrosis, especially in premature newborn infants who have low glutathione levels.

Postnatal Gender-dependent Maturation of Cellular Cysteine Uptake

Background. In view of the functional capacity of glutathione synthesis in premature infants, and because the availability of cysteine is one the rate limiting steps in glutathione synthesis, we hypothesized that the low glutathione levels in premature infants may be due to immaturity of the active cellular uptake of cysteine. Objective. To document in cells from newborn infants the effect of maturity and gender on cysteine uptake and consequently on glutathione levels. Methods. Incorporation of L‐[35S] cysteine was measured in leukocytes from cord blood and from tracheal aspirates (TAC) of newborn infants of varying (gestational as well as postnatal) ages and gender. Cysteine uptake was correlated with glutathione in TAC. Results. The maturity of newborn girls positively influences cysteine uptake, which is responsible for 78% of the variation in their glutathione content. However, in newborn boys, gestational and postnatal ages did not influence the cysteine uptake. Discussion. Cysteine uptake appears to be the limiting step explaining the reported gender-related differences in glutathione as well as the low levels of this central antioxidant found in premature infants. The immature cysteine uptake found in cells from premature infants raises questions about the bioavailability of this conditionally essential amino acid in regimens of parenteral nutrition for human neonates.

The mode of administration of total parenteral nutrition and nature of lipid content influence the generation of peroxides and aldehydes

BACKGROUND & AIMS The absence of light protection of neonatal total parenteral nutrition (PN) contributes to the generation of 4-hydroxynonenal and peroxides. 4-Hydroxynonenal is suspected to be involved in PN-related liver complications. AIMS To find a practical modality to reduce 4-hydroxynonenal in PN and assess in vivo the impact of PN containing low 4-hydroxynonenal concentration. METHODS Six modalities of delivering PN were compared for the in vitro generation of peroxides and 4-hydroxynonenal: 1) MV-AA-L: light-protected (-L) solution containing multivitamin (MV) mixed with amino acids + dextrose (AA); 2) MV-AA+L: MV-AA without photo-protection (+L); 3) MV-LIP+L: MV mixed with lipid emulsion (LIP). LIP was a) Intralipid20%(®) or b) Omegaven(®). Hepatic markers of oxidative stress (glutathione, F(2α)-isoprostanes, GS-HNE) and inflammation (mRNA of TNF-α and IL-1) were measured in newborn guinea pigs infused during 4-days with MV-AA+L compounded with Intralipid20%(®) or Omegaven(®). RESULTS Hydroperoxides and 4-hydroxynonenal were the lowest in MV-AA-L and the highest in MV-LIP+L. MV-AA+L with Omegaven(®) was associated with the lowest levels of markers of oxidative stress and inflammation. CONCLUSION Compared to Intralipid20%(®), Omegaven(®) reduces oxidative stress associated with PN and prevents liver inflammation. These findings offer an alternative strategy to light protection of PN, which in the clinical setting is a cumbersome modality.

Modulation of apo A-IV transcript levels and synthesis by n-3, n-6, and n-9 fatty acids in CACO-2 cells.

It has been postulated that apolipoprotein (apo) A‐IV plays various significant roles in lipid transport and lipoprotein metabolism. Although it is controlled by fat feeding, so far little else is known about its regulation by specific fatty acids. In this study, we focused on the modulation of apo A‐IV mRNA levels, mass, and biogenesis by mono‐ and polyunsaturated fatty acids (FA) in the human intestinal Caco‐2 cell line. In confluent cells incubated with 1 mM oleic (n‐9), linoleic (n‐6), α‐linolenic (n‐3), or docosahexaenoic (n‐3) acids for a long‐term period, both apo A‐IV protein levels and de novo synthesis were increased. The induction resulted from the up‐regulation of apo A‐IV mRNA transcripts. In contrast, an inhibitory effect was evident with short‐term incubation. FA chain length and degree of unsaturation had little effect altering apo A‐IV transcript and biogenesis. These data offer evidence that isolated fatty acids regulate gene expression and the production of apo A‐IV in the enterocyte. J. Cell. Biochem. 75:73–81, 1999.