Abstract
Alpha-ketoglutarate, a key intermediate in the Krebs cycle, has been reported to benefit intestinal health. We tested whether alpha-ketoglutarate can protect intestinal cells against hydrogen peroxide induced damage and aimed to reveal the underlying mechanism. Intestinal porcine epithelial cell line J2 were cultured in Dulbecco’s Modified Eagle Medium-High glucose with or without alpha-ketoglutarate and hydrogen peroxide. Cell viability, proliferation, mitochondrial respiration, mitochondrial membrane potential, antioxidant function, apoptosis and mitochondrial-dependent apoptotic pathways were determined. Our experiments demonstrated that, first, exposure to 100μM hydrogen peroxide decreased cell viability, DNA synthesis, mitochondrial respiration and antioxidant function, and increased apoptosis. Second, 2mM alpha-ketoglutarate addition attenuated hydrogen peroxide-induced cell cycle arrest, and improved cell viability, DNA synthesis, mitochondrial respiration and antioxidant function. Third, alpha-ketoglutarate enhanced tricarboxylic acid cycle activity, mitochondrial respiration, and decrease the intracellular content of reactive oxygen species. Finally, alpha-ketoglutarate stabilized the mitochondrial membrane potential, increased the ratio of Bcl-2/Bax, decreased the release of cytochrome c and activation of caspase-3, thereby prevented cell apoptosis. Altogether, we proposed that alpha-ketoglutarate protects intestinal cells against hydrogen peroxide-induced damage partly via mitochondria dependent pathway.
Highlights
The intestinal epithelium is the most important internal barrier against the invasion of pathogenic bacteria [1], and functions as an important part of the first line of defense [2]
First, exposure to 100μM hydrogen peroxide decreased cell viability, DNA synthesis, mitochondrial respiration and antioxidant function, and increased apoptosis
The results indicated that H2O2 decreased IPEC-J2 cell viability in a dose-dependent manner; and when itsconcentration was increased to 100 μM, H2O2 showed significant inhibitory effects on cell viability (P < 0.05) (Figure 1A)
Summary
The intestinal epithelium is the most important internal barrier against the invasion of pathogenic bacteria [1], and functions as an important part of the first line of defense [2]. In swine production, weaning stress has been demonstrated to be associated with gastrointestinal disorders such as changes in intestinal barrier function and absorption, leading to diarrhea and increased disease susceptibility [3, 4]. Oxidative stress refers to the imbalance of oxidation and antioxidation, which leading to accumulation of oxidation intermediates including superoxide ion (O2-), hydroxyl radical (OH) and H2O2 [6]. It causes intestinal disorders possibly via [2, 7, 8] : 1) disrupting the structures of the intestinal barrier, 2) influencing mitosis or apoptosis of intestinal epithelial cells, or 3) distorting the differentiation potential of epithelial cells. Maintaining a balanced redox status is essential for ensuring intestinal health [9]
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