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    L-Arginine Alleviates Hydrogen Peroxide-Induced Oxidative Damage in Ovine Intestinal Epithelial Cells by Regulating Apoptosis, Mitochondrial Function, and Autophagy
    (Oxford Univ Press, 2021) Zhang, Hao; Liu, Xiaoyun; Fan, Yaotian; Yu, Yin; Loor, Juan J.; Elsabagh, Mabrouk; Peng, Along
    Background: Previous studies demonstrated that dietary L-arginine (Arg) alters the equilibrium between reactive oxygen species (ROS) generation and biological defenses to resist oxidant-induced toxicity. Whether supplying Arg can protect ovine intestinal epithelial cells (OIECs) from hydrogen peroxide (H2O2)-induced oxidative damage is unclear. Objectives: The current study aimed to examine the effect of Arg on mitophagy, mitochondrial dysfunction, and apoptosis induced by H2O2 in OIECs. Methods: The OIECs were incubated in Arg-free DMEM supplemented with 100 AM Arg (CON) or 350 AM Arg (ARG) alone or with 150 mu M H2O2 (CON + H2O2, ARG + H2O2) for 24 h. Cellular apoptosis, mitochondrial function, autophagy, and the related categories of genes and proteins were determined. All data were analyzed by ANOVA using the general linear model procedures of SAS (SAS Institute) for a 2 x 2 factorial design. Results: Relative to the CON and ARG groups, H2O2 administration resulted in 44.9% and 26.5% lower (P < 0.05) cell viability but 34.7% and 61.8% greater (P < 0.05) ROS concentration in OIECs, respectively. Compared with the CON and CON + H2O2 groups, Arg supplementation led to 40.7% and 28.8% lower (P < 0.05) ROS concentration but 14.9%-49.0% and 29.3%-64.1% greater (P < 0.05) mitochondrial membrane potential, relative mitochondrial DNA content, and complex (I-IV) activity in OIECs, respectively. Compared with the CON and CON + H2O2 groups, Arg supplementation led to 33.9%-53.1% and 22.4%-49.1 % lower (P < 0.05) mRNA abundance of proapoptotic genes, respectively. Relative to the CON and CON + H2O2 groups, Arg supplementation resulted in 33.0%-59.2% and 14.6%-37.7% lower (P< 0.05) abundance of proapoptotic, mitophagy, and cytoplasmic cytochrome c protein, respectively. Conclusions: Supply of Arg protects OIECs against H2O2-induced damage partly by improving mitochondrial function and alleviating cellular apoptosis and autophagy.
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    L-Arginine inhibits hydrogen peroxide-induced oxidative damage and inflammatory response by regulating antioxidant capacity in ovine intestinal epithelial cells
    (Taylor & Francis Ltd, 2021) Zhang, Hao; Zhang, Ying; Liu, Xiaoyun; Elsabagh, Mabrouk; Yu, Yin; Peng, Along; Dai, Sifa
    Little is known how L-arginine (Arg) affects the ovine intestinal epithelial cells (IOECs) redox status induced by hydrogen peroxide (H2O2)(.) This study aimed to examine the impact of Arg on IOECs subjected to H2O2-induced oxidative damage, intestinal barrier injury, and inflammatory response. The IOECs were incubated for 16 h then classified as four groups (n = 6/group) and cultured in corresponding media including (1) control (CON) group, in which IOECs were cultured in Arg-free Dulbecco's modified Eagle's F12 Ham medium (DMEM) containing 100 mu M Arg; (2) Arg group, in which IOECs were cultured in Arg-free DMEM containing 350 mu M Arg; (3) H2O2 group, in which IOECs were cultured in CON group plus 150 mu M H2O2; (4) Arg + H2O2 group, in which IOECs were cultured in Arg group plus150 mu M H2O2. After culturing for 24 h in media, some characteristics of cells in the four groups were measured. Arg administration decreased the H2O2-induced reactive oxygen species (ROS) production compared with the H2O2 group (p < .05). Compared with H2O2, adding Arg to H2O2 increased (p < .05) transepithelial electrical resistance (TEER), but decreased (p < .05) tumour necrosis factor alpha (TNF-alpha) within the IOECs. Compared with H2O2, adding Arg to H(2)O(2-)induced injured IOECs increased (p < .05) glutathione peroxidase 1 (GPx1), zonula occludens-1 (ZO-1), and epithelial nitric oxide (NO) synthase (eNOS) protein levels, and decreased (p < .05) TNF-alpha levels within cells. Arg inhibits H2O2-induced oxidative damage, intestinal barrier injury, and inflammatory response by NO pathway within IOECs.