Effects of Vitamin E Supplementation on Antioxidant, Inflammatory Biomarker, and Cell Viability of Peripheral Blood Mononuclear Cells from Japanese Black Calves with or without Lipopolysaccharide Stimulation

Abstract

The purpose of this study was to determine the effects of vitamin E supplementation on antioxidant, inflammatory status, and cell viability of peripheral blood mononuclear cells. Eighteen clinically healthy Japanese Black calves were used in this study. The peripheral blood mononuclear cells (PBMCs) were isolated from their venous blood. PBMCs were cultured with vitamin E (vitamin E group) or without vitamin E (control group), and stimulated with or without lipopolysaccharide (LPS). As a result, the total antioxidant capacities, which are the reducing power of components from Fe 31 to Fe 21 , in the cell culture supernatant with or without LPS stimulation were significantly higher in the vitamin E group than that in the control group (p,0.05 or p,0.01, respectively). Tumor necrosis factor alpha in the cell culture supernatant with LPS stimulation was significantly lower in the vitamin E group than in the control group (p,0.01). The viability of cells cultured with LPS stimulation was significantly higher in the vitamin E group than in the control group (p,0.05). These results suggested that vitamin E might be related to the antioxidant, anti-inflammatory, and cell viability of PBMCs obtained from calves.

1,25‐Dihydroxyvitamin D3 potentiates the innate immune response of peripheral blood mononuclear cells from Japanese Black cattle

“…The cells were treated with or without 1 μg/ml LPS ( Escherichia coli O111; Sigma Aldrich, United States) and 10 nM 1,25(OH) 2 D 3 , also known as Calcitriol (Fujifilm Wako, Japan) at 37°C for 24 and 72 h in a humidified 5% CO 2 atmosphere. The concentration of LPS and 1,25(OH) 2 D 3 was determined in accordance with previous reports (Otomaru et al, 2022; Waters et al, 2001; Wherry et al, 2022). After the treatments, live cells were counted with trypan blue (0.5% trypan blue stain, Nacalai Tesque, Japan), and the cell viability was calculated by dividing the number of cells after 24 and 72 h treatment by that of 0 h. It was confirmed that almost all cells were alive at the 0 h treatment.…”

“…Blood was collected from the jugular vein once each from 21 calves between 24 and 32 weeks of age using vacutainer heparin tubes. PBMCs were purified as previously described (Maeda et al, 2011; Otomaru et al, 2022, 2023). Briefly, 10 ml of heparinized blood was diluted twice in phosphate‐buffered saline (PBS), placed above the separation medium solution (Lymphocyte Separation Medium 1077, Immuno‐Biological Laboratories, Japan; specific gravity 1.077), and then centrifuged at 400× g for 60 min at 15°C.…”

“…PBMCs were purified as previously described (Maeda et al, 2011;Otomaru et al, 2022Otomaru et al, , 2023. Briefly, 10 ml of heparinized blood was diluted twice in phosphate-buffered saline (PBS), placed above the separation medium solution (Lymphocyte Separation Medium 1077, Immuno-Biological Laboratories, Japan; specific gravity 1.077), and then centrifuged at 400Âg for 60 min at 15 C. Mononuclear cells at the interface were collected and then washed twice with PBS by centrifugation at 400Âg for 5 min.…”

Effects of Vitamin C Supplementation to Bovine Peripheral Blood Mononuclear Cells on Antioxidant and Inflammatory Biomarker Status and Cell Viability with or without Lipopolysaccharide Stimulation

“…Studies reported that in vitro vitamin C supplementation to human blood leukocytes reduced the production of TNF-α and IL-6 (Hartel et al 2004), and vitamin C supplementation to human blood plasma or porcine kidney cells increased antioxidant capacity (Bleilevens et al 2019, Hong et al 2002. There have been reports about in vitro vitamin E or anti-inflammatory agent supplementation to peripheral blood mononuclear cells (PBMCs) of cattle (Maeda et al 2011, Otomaru et al 2022. However, only few in vitro studies have investigated vitamin C supplementation to bovine tissues or cells.…”