Resveratrol Ameliorates Intestinal Damage Challenged With Deoxynivalenol Through Mitophagy in vitro and in vivo

Abstract

Deoxynivalenol (DON) reduces growth performance and damage intestinal function, and resveratrol (RES) has positive effects on growth performance and intestinal function. The purpose of this study was to investigate the protective mechanism of RES in vitro and vivo challenged with DON. The results showed that dietary supplementation with DON significantly increase the mRNA expression levels of mitophagy- related genes, and protein level for PINK1, Parkin, Beclin-1, Lamp, Atg5, Map1lc, Bnip3, Fundc1, Bcl2l1 and SQSTMS1 (P < 0.05), while supplementation with both RES and DON decreased those indexes in the ileum. Besides DON significantly decreased protein level for Pyruvate Dehydrogenase, Cytochrome c, MFN1, OPA1, and PHB1 (P < 0.05), while supplementation with both RES and DON increased protein level for PHB1, SDHA, and VDAC in the ileum. Moreover, in vitro, we found that DON significantly decreased mitochondrial respiration (P < 0.05), while RES + DON increased the rate of spare respiratory capacity. Also, DON significantly decreased total NAD and ATP (P < 0.05), while RES + DON increased the total NAD and ATP. These results indicate that RES may ameliorates the intestinal damage challenged with deoxynivalenol through mitophagy in weaning piglets.

Keywords: deoxynivalenol; intestinal function; mitophagy; piglets; resveratrol.

Figure 1

Dietary supplementation with RES alleviated the negative effects on mRNA expression levels of mitophagy-related genes challenged with DON. Data were expressed as means ± SEM of at least three independent experiments. (A,B) Values with different letters are significantly different (P < 0.05).

Figure 2

Dietary supplementation with RES increases the expression of mitophagy-related genes. Cells were treated with 0 (NC) or 0.5 μmol/L DON and 0 or 15 μM RES, respectively. Data were expressed as means ± SEM of at least three independent experiments. (A,B) Values with different letters are significantly different (P < 0.05).

Figure 3

Dietary supplementation with RES affects the expression of mitochondrial related genes. Cells were treated with 0 (NC) or 0.5 μmol/L DON and 0 or 15 μM RES, respectively. Data were expressed as means ± SEM of at least three independent experiments. (A,B) Values with different letters are significantly different (P < 0.05).

Figure 4

RES improved the negative effect on mitochondrial respiration by DON in vitro. (A) Schematic and (B) oxygen consumption rate (OCR) assessed by extracellular flux analysis. OCR was measured under basal conditions followed by the sequential addition of oligomycin (0.5 μM), FCCP (1 μM), rotenone (1 μM), or antimycin A (1 μM). Each data point represents an OCR measurement. (C) Individual parameters for basal respiration, proton leak, maximal respiration, spare respiratory capacity, nonmitochondrial respiration, and ATP production were determined. Cells were treated with 0 (NC) or 0.5μmol/L or 1μmol/L DON and 0 or 15μM RES, respectively. Data were expressed as means ± SEM of at least three independent experiments. (A–C) Values with different letters are significantly different (P < 0.05).

Figure 5

(A,B) Effect of DON and RES on Total NAD and ATP. Cells were treated with 0 (NC) or 0.5 μmol/L or 1 μmol/L DON and 0 or 15 μM or 20 μM RES, respectively. Data were expressed as means ± SEM of at least three independent experiments. *Values with different letters are significantly different (P < 0.05), *P < 0.05, **P < 0.01, ***P < 0.001.

Figure 6

RES did not relieve injury on mitochondrial respiration caused by DON when knockout Atg5. (A) Schematic and (B) oxygen consumption rate (OCR) assessed by extracellular flux analysis. OCR was measured under basal conditions followed by the sequential addition of oligomycin (0.5 μM), FCCP (1 μM), rotenone (1 μM), or antimycin A (1 μM). Each data point represents an OCR measurement. (C) Individual parameters for basal respiration, proton leak, maximal respiration, spare respiratory capacity, nonmitochondrial respiration, and ATP production were determined. Cells were treated with 0 (NC) or 0.5 μmol/L or 1 μmol/L DON and 0 or 15 μM RES, respectively. Data were expressed as means ± SEM of at least three independent experiments. (A–C) Values with different letters are significantly different (P < 0.05).