白藜芦醇对急性热应激条件下鸭肝抗氧化能力和细胞凋亡的影响

doi:10.11843/j.issn.0366-6964.2023.01.022

摘要/Abstract

摘要： 旨在探索饲粮中添加白藜芦醇对急性热应激条件下山麻鸭肝抗氧化能力和细胞凋亡的影响，确定添加白藜芦醇对山麻鸭肝组织SIRT1信号通路、抗氧化酶系统及凋亡相关基因的作用。试验选取120只60日龄体重相近且健康的雌性山麻鸭，随机分为2组，每组60只，既对照组(C，饲喂基础饲粮)和白藜芦醇组(RES，饲喂添加400mg·kg^-1白藜芦醇的基础饲粮)。两组动物均在常温(24℃±2℃)下饲养，15d后，两组动物体重无明显差异。随后分别将每组动物随机分为3组，每组20只，在39℃环控仓中分别放置0、30、60min。热应激处理后，采集鸭血清样本及肝组织，应用qRT-PCR、Western blot、TUNEL技术对SIRT1信号通路及凋亡相关基因的mRNA、蛋白水平和细胞凋亡数进行检测。采用生化法检测血清及肝中的氧化指标。结果显示，与对照组相比，白藜芦醇组HSP70和HSP90 mRNA表达有所升高。同时，白藜芦醇激活了SIRT1途径，增加了下游PGC-1α、TFAM mRNA表达，显著增加了热应激60min条件下Nrf1、Nrf2核蛋白的表达。白藜芦醇提高了血清中T-AOC水平，在热应激60min时，白藜芦醇显著提高了血清中SOD、CAT、AIHR的水平并显著降低MDA的含量。在同一热应激条件下，RES组肝中T-AOC、CAT、GSH-Px均高于C组，RES组MDA的含量显著低于C组。在急性热应激条件下，RES组的肝细胞凋亡数显著低于C组，且白藜芦醇降低了Bax、Bak1、Bax/Bcl-2 mRNA的表达量，同时增加了Bcl-2蛋白的表达量，降低了Bax、Caspase3、Bax/Bcl-2蛋白的表达量。综上所述，饲料中添加400mg·kg^-1白藜芦醇可以激活鸭肝SIRT1信号通路，使促凋亡基因的表达量下降，抗凋亡基因的表达量增加。白藜芦醇可提高鸭肝的抗氧化能力和抗细胞凋亡的能力，能够改善急性热应激对鸭肝的氧化损伤。

关键词: 白藜芦醇, 热应激, 抗氧化, 凋亡

Abstract: This study was conducted to explore the effects of dietary resveratrol supplementation on liver anti-oxidant capacity and anti-apoptosis of Shanma ducks under acute heat stress, and to determine the effects of resveratrol supplementation on SIRT1 signaling pathway, antioxidant enzyme system and apoptosis related genes in the liver of Shanma ducks. A total of 120 healthy female Shanma ducks with similar weight at 60 days old were randomly divided into two groups, 60 in each group. The control (C) group was fed a basic diet, and the resveratrol (RES) group was fed a basic diet supplemented with 400 mg·kg^-1 resveratrol. Animals in two groups were kept at a temperature of (24±2) ℃. After 15 days, there was no significant difference in the weight of the animals between the two groups. Then the animals in each group were randomly divided into 3 groups,20 in each group, and placed in an artificial climate room at 39 ℃ for 0, 30, and 60 min, respectively. After heat stress treatment, duck serum samples and liver tissues were collected, and the mRNA, protein level and apoptosis number of SIRT1 signal pathway and apoptosis related genes were detected using qRT-PCR, Western blot and TUNEL techniques. Biochemical method was adopted to detect the oxidation index in serum and liver. The results showed that the mRNA expression of HSP70and HSP90 in RES group was higher than those in control group. At the same time, resveratrol activated the SIRT1 pathway and increased the mRNA expressions of downstream PGC-1α and TFAM, and significantly increased the nuclear proteins expression of Nrf1 and Nrf2 under heat stress for 60 min. Resveratrol increased the levels of T-AOC in serum. Resveratrol significantly increased the levels of SOD, CAT, AIHR and decreased the content of MDA under heat stress for 60 min，and under the same heat stress condition， the T-AOC, CAT and GSH-Px in liver of RES group were higher than those of C group, while MDA content in RES group was lower than that in C group. Under the condition of acute heat stress, the number of hepatocyte apoptosis in RES group was significantly lower than that in C group, and resveratrol decreased the mRNA expression of Bax, Bak1, Bax/Bcl-2, increased the protein expression of Bcl-2, and decreased the protein expression of Bax, Caspase3, Bax/Bcl-2. In conclusion, the addition of 400 mg·kg^-1 resveratrol to the diet can activate the SIRT1 signaling pathway in duck liver, reduce the expression of pro-apoptotic genes and increase the expression of anti-apoptotic genes. Resveratrol can improve the anti-oxidant capacity and anti-apoptosis ability of duck liver, and can improve the oxidative damage of duck liver caused by acute heat stress.

Key words: resveratrol, heat stress, antioxidant, apoptosis

中图分类号:

S834.4

周婉婷, 杨晨, 彭翠甜, 付新亮, 钟焯华, 许丹宁, 黄运茂, 田允波, 刘文俊. 白藜芦醇对急性热应激条件下鸭肝抗氧化能力和细胞凋亡的影响[J]. 畜牧兽医学报, 2023, 54(1): 239-251.

ZHOU Wanting, YANG Chen, PENG Cuitian, FU Xinliang, ZHONG Zhuohua, XU Danning, HUANG Yunmao, TIAN Yunbo, LIU Wenjun. Effects of Resveratrol on Anti-oxidation and Anti-apoptosis of Hepatocytes of Ducks on Exposure to Acute Heat Stress[J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(1): 239-251.

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