Effects of Human Milk and Spermine on Hydrogen Peroxide‐Induced Oxidative Damage in IEC‐6 Cells

Shoji, Hiromichi; Oguchi, Satoshi; Fujinaga, Shuichiro; Shinohara, Koichi; Kaneko, Kazunari; Shimizu, Toshiaki; Yamashiro, Yuichiro

doi:10.1097/01.mpg.0000176180.89261.bf

Cited by 22 publications

(20 citation statements)

References 40 publications

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“…Further studies are required to elucidate the mechanisms involved. Cell viability of IEC-6 cells with induced oxidative damage was increased by the addition of human colostrum, but not cow milk or infant formulas, and the increase was attributed to the polyamine spermine reducing the oxidative stress [41]. Polyamines are small cationic molecules that are found in human milk and are required for intestinal growth and development [42].…”

Section: Discussionmentioning

confidence: 99%

Goat milk with and without increased concentrations of lysozyme improves repair of intestinal cell damage induced by enteroaggregative Escherichia coli

et al. 2012

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BackgroundEnteroaggregative Escherichia coli (EAEC) causes diarrhea, malnutrition and poor growth in children. Human breast milk decreases disease-causing bacteria by supplying nutrients and antimicrobial factors such as lysozyme. Goat milk with and without human lysozyme (HLZ) may improve the repair of intestinal barrier function damage induced by EAEC. This work investigates the effect of the milks on intestinal barrier function repair, bacterial adherence in Caco-2 and HEp-2 cells, intestinal cell proliferation, migration, viability and apoptosis in IEC-6 cells in the absence or presence of EAEC.MethodsRat intestinal epithelial cells (IEC-6, ATCC, Rockville, MD) were used for proliferation, migration and viability assays and human colon adenocarcinoma (Caco-2, ATCC, Rockville, MD) and human larynx carcinoma (HEp-2, ATCC, Rockville, MD) cells were used for bacterial adhesion assays. Goats expressing HLZ in their milk were generated and express HLZ in milk at concentration of 270 μg/ml . Cells were incubated with pasteurized milk from either transgenic goats expressing HLZ or non-transgenic control goats in the presence and absence of EAEC strain 042 (O44:H18).ResultsCellular proliferation was significantly greater in the presence of both HLZ transgenic and control goat milk compared to cells with no milk. Cellular migration was significantly decreased in the presence of EAEC alone but was restored in the presence of milk. Milk from HLZ transgenic goats had significantly more migration compared to control milk. Both milks significantly reduced EAEC adhesion to Caco-2 cells and transgenic milk resulted in less colonization than control milk using a HEp-2 assay. Both milks had significantly increased cellular viability as well as less apoptosis in both the absence and presence of EAEC.ConclusionsThese data demonstrated that goat milk is able to repair intestinal barrier function damage induced by EAEC and that goat milk with a higher concentration of lysozyme offers additional protection.

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Section: Discussionmentioning

confidence: 99%

Goat milk with and without increased concentrations of lysozyme improves repair of intestinal cell damage induced by enteroaggregative Escherichia coli

et al. 2012

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“…The level of urinary 8-hydroxy-2=-deoxyguanosine (a marker of oxidative DNA damage) excretions of breast-fed infants was significantly lower than that of formula-fed infants in 1 mo of age (4). HM-treated IEC-6 cells showed a significantly higher survival rate after H 2 O 2 -induced oxidative damage than cells treated with infant formula (5). Despite these findings, it is still unclear which components of HM play the most important antioxidative roles in the GIT of infants.…”

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confidence: 99%

Effects of Iron-Unsaturated Human Lactoferrin on Hydrogen Peroxide-Induced Oxidative Damage in Intestinal Epithelial Cells

et al. 2007

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Human milk (HM) contains various bioactive antioxidants. Lactoferrin (Lf) has been assumed to be one of the major antioxidants in HM. We examined the antioxidative properties of iron-unsaturated human Lf (apo-hLf, the major form of Lf in HM) in two intestinal epithelial cell lines: (1) An intestinal epithelial cell line (IEC-6) were preincubated for 24 h with either 50 g/mL of apo-hLf, iron-saturated human Lf (holo-hLf), iron-unsaturated bovine transferrin (apo-bTf), or 800 ng/mL of the iron-chelating compound deferoxamine (DFX), followed by hydrogen peroxide (H 2 O 2 ) challenge to induce oxidative stress. Survival rates were significantly higher in the cells preincubated with apo-hLf and DFX than those preincubated with holo-hLf. (2) Caco-2 cells were preincubated with or without apo-hLf for 24 h, followed by an H 2 O 2 challenge. Intracellular oxidative stress was assessed by a fluorescent probe, 2=,7=-dichlorodihydrofluorescein diacetate (DCF-DA). Fluorescent intensity of cell images and cell homogenates was significantly lower in the cells preincubated with apo-hLF than those preincubated without apo-hLF. Our study indicates that apo-hLf alleviates H 2 O 2 -induced oxidative damage in intestinal cells due to the iron-chelating capacity. Therefore, Lf in HM may act as an antioxidant in the gastrointestinal tract (GIT). O xidative stress occurs when there is an imbalance between the amount of antioxidant properties and reactive oxygen species. In the neonatal period, especially in preterm infants injury to the GIT by oxidative stress is thought to be involved in the pathogenesis of serious diseases such as necrotizing enterocolitis (1,2).HM is the ideal food during infancy and is known to contain various types of bioactive substances, some of which are reported to be antioxidants (3). We demonstrated the antioxidative effects of HM in two previous studies. The level of urinary 8-hydroxy-2=-deoxyguanosine (a marker of oxidative DNA damage) excretions of breast-fed infants was significantly lower than that of formula-fed infants in 1 mo of age (4). HM-treated IEC-6 cells showed a significantly higher survival rate after H 2 O 2 -induced oxidative damage than cells treated with infant formula (5). Despite these findings, it is still unclear which components of HM play the most important antioxidative roles in the GIT of infants.Lf is an iron-binding glycoprotein that belongs to the transferrin (Tf) family. Although it has been identified in many secretions from various species, it mainly exists in mammalian milk and colostrum. The amino acid compositions of bovine Lf and human Lf show 69% sequence homology (6). Lf appears to play a role in many biologic processes, including the proliferation and differentiation of enterocytes (7), regulation of iron absorption (8), antimicrobial activities (9), immunomodulation and anti-inflammatory responses (10), and antivirus or anticancer activities (11). Lf is considered to be an antioxidant because of its ability to bind two atoms of iron, which is important in t...

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“…6,7 In vitro, human breast milk antioxidants were observed to decrease hydrogen peroxide-induced oxidative damage in intestinal epithelial cells. 3,4 Antioxidant properties of human breast milk support the natural antioxidant barrier of neonates and infants. Lower levels of oxidative stress are observed in breastfed infants when compared with those receiving infant formulas, 8,9 including the antioxidant-enriched formulas.…”

Section: Introductionmentioning

confidence: 99%

“…This antioxidant barrier includes enzymes as well as non-enzymatic components. Human breast milk contains either non-enzymatic antioxidants (among others coenzyme Q, lactoferrin and vitamins A, E and C) [2][3][4][5] or antioxidant enzymes. 6,7 In vitro, human breast milk antioxidants were observed to decrease hydrogen peroxide-induced oxidative damage in intestinal epithelial cells.…”

Section: Introductionmentioning

confidence: 99%

Maternal smoking decreases antioxidative status of human breast milk

Zagierski

Szlagatys-Sidorkiewicz

Jankowska

et al. 2011

J Perinatol

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Objective: To evaluate the influence of maternal smoking on antioxidative capacity and intensity of oxidative damage in breast milk.Study Design: The study group (n ¼ 30) was comprised of postpartum women who declared smoking more than five cigarettes per day during pregnancy and lactation (confirmed by the urinalysis of cotinine concentration), and their newborns. Control group included 29 nonsmoking postpartum women and their newborns. Colostrum samples were collected on the 3rd day after delivery and breast milk samples between the 30th and the 32nd day after delivery. Morning maternal and neonatal urine samples were obtained on the day of the mature milk sampling. Isoprostane concentrations in colostrum/mature milk and urine were determined immunoenzymatically. Total Antioxidant Status (TAS) of colostrum/breast milk was determined by Rice-Evans and Miller method.Result: Colostrum TAS in smokers was significantly lower than in nonsmokers (P ¼ 0.006). In both groups, the TAS of mature milk was higher compared with colostrum, but significant differences were observed amongst smokers only (P ¼ 0.001). In smokers the isoprostane concentration of mature milk was significantly higher than the colostrum concentration (P ¼ 0.001). Significant inverse correlation between maternal urinary isoprostane concentration and the TAS of mature breast milk was observed in smokers (R ¼ À0.525, P ¼ 0.023), but not in non-smokers (R ¼ 0.161, P ¼ 0.422).Conclusion: This study revealed that maternal smoking triggers harmful effects on an infant by impairing pro-oxidant-antioxidant balance of breast milk.

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Effects of Human Milk and Spermine on Hydrogen Peroxide‐Induced Oxidative Damage in IEC‐6 Cells

Cited by 22 publications

References 40 publications

Goat milk with and without increased concentrations of lysozyme improves repair of intestinal cell damage induced by enteroaggregative Escherichia coli

Goat milk with and without increased concentrations of lysozyme improves repair of intestinal cell damage induced by enteroaggregative Escherichia coli

Effects of Iron-Unsaturated Human Lactoferrin on Hydrogen Peroxide-Induced Oxidative Damage in Intestinal Epithelial Cells

Maternal smoking decreases antioxidative status of human breast milk

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