最新录用：外源性γ-氨基丁酸对仔猪抗氧化应激作用的效果及机制

doi:10.3864/j.issn.0578-1752.210595

摘要/Abstract

摘要： 【目的】研究外源性γ-氨基丁酸（GABA）抵抗仔猪氧化应激的作用效果，及海马神经元GABA受体调节凋亡信号通路在其中可能的介导作用，为GABA作为动物应激调节剂的应用提供科学依据。【方法】基于成功构建的仔猪活体和大鼠海马神经元氧化应激模型，考察外源性GABA对仔猪血清及海马组织中氧化/抗氧化相关指标、仔猪日增重、脑海马GABA受体以及大鼠海马神经元中GABA受体及凋亡信号通路相关基因表达水平的影响。【结果】低、中、高浓度的GABA灌喂组（LD+OS； MD+OS； HD+OS）仔猪血清的MDA含量均极显著低于氧化应激（OS）组(P<0.01)，而GSH水平均极显著高于OS组(P<0.01)，同时HD+OS 组仔猪血清T-AOC水平极显著高于OS组和对照组(P<0.01)；且高浓度(100 mg·kg^-1 BW)GABA降低仔猪血清MDA含量和增加GSH水平的幅度均高于低浓度(20mg·kg^-1 BW)和中浓度(60 mg·kg^-1 BW)GABA。因此，后续研究仅考察100 mg·kg^-1 BW GABA的作用效果及其抗氧化应激的机制。 OS组仔猪0-7、8-14和0-28日龄的日增重极显著低于对照组(P<0.01)，而HD+OS组0-7、8-14和0-28日龄仔猪的日增重极显著高于OS组(P<0.01)；OS组仔猪15-28日龄的日增重显著低于对照组(P <0.05)，HD+OS组15-28日龄的日增重极显著高于OS组(P <0.01)。以上结果表明，100 mg·kg^-1 BW GABA的灌喂极显著地增加了仔猪的日增重。对照组、OS组和HD+OS组在前、中、后期的腹泻率均无显著差异(P >0.05)。氧化应激组海马组织的MDA含量极显著高于对照组和HD+OS组 (P <0.01)，而T-AOC和GSH的水平均极显著低于另外两组(P <0.01)，表明GABA能提高仔猪海马组织抗氧化能力。HD+OS组的海马组织GABA_A和GABA_B受体水平均极显著高于对照组和氧化应激组(p<0.01)，表明GABA提高了海马组织GABA_A和GABA_B的水平。氧化应激组脑海马Bcl-2蛋白水平显著低于对照组(p<0.05)，Bax和Caspase-3蛋白水平极显著高于对照组(P <0.01)。HD+OS组的Bcl-2蛋白水平极显著高于氧化应激组（P <0.01），Bax蛋白水平极显著低于氧化应激组（P <0.01），Caspase-3蛋白水平显著低于氧化应激组（P <0.05）。与此一致的是，氧化应激组、GABA+OS+Picrotoxin组和GABA+OS+CGP54626组大鼠海马神经元的Bax和Caspase-3蛋白水平均显著高于对照组和GABA+OS组（P <0.05），表明GABA缓解了氧化应激状态下海马神经元的损伤，而GABA受体抑制剂的添加阻挡了GABA的抗应激损伤作用。【结论】GABA降低了仔猪海马的氧化应激水平，而GABA抗应激的作用机制可能与其降低凋亡蛋白基因的表达相关，而GABA_A和GABA_B受体介导了该过程。

关键词: GABA, 仔猪, 氧化应激, 受体, 凋亡

Abstract: 【Objective】The purpose of this paper is to study the effect of exogenous γ-aminobutyric acid against oxidative stress in piglets, and the possible mediating role of hippocampal neuron γ-aminobutyric acid receptors regulating apoptosis signaling pathways in it, which will provide scientific basis for the application of γ-aminobutyric acid as animal stress regulator. 【Method】 Based on the successful establishment of oxidative stress model in piglet and rat hippocampal neurons, the effects of exogenous GABA on piglets under oxidative stress, including oxidative / antioxidant related indexes, growth performance related indexes and GABA receptor in serum and hippocampus, along with the GABA receptor and apoptosis signal pathway related indexes in hippocampal neurons were investigated. 【Result】 The serum MDA levels of piglets in the low, medium, and high concentration GABA feeding groups (LD+OS; MD+OS; HD+OS) were extremely significantly lower than those in the oxidative stress (OS) group (P <0.01), the GSH level was extremely significantly higher than that in the OS group (P <0.01), and the T-AOC level in the HD+OS group was extremely significantly higher than that in the OS group and control group (P <0.01. And the high concentration (100mg·kg^-1 BW) of GABA more decreased the MDA level and more increased the GSH level in serum of piglets than those of the low concentration (20mg·kg^-1 BW) and middle concentration (60mg·kg^-1 BW) of GABA. Therefore, the follow-up study only investigated the anti-oxidative stress effect and mechanism of 100 mg·kg^-1 BW. The daily gains of piglets at 0-7 days, 8-14 days and 0-28 days in the OS group were extremely significantly lower than those in the control group (P <0.01)，the daily gains of piglets at 0-7 days, 8-14 days and 0-28 days in the HD+OS group were extremely significantly higher than those in the OS group (P <0.01), the daily gain of piglets aged 15-28 days in the OS group was significantly lower than that in the control group (P <0.05)，the daily gain of piglets aged 15-28 days in the HD+OS group was extremely significantly higher than that in the OS group (P <0.01). The above results showed that the 100 mg·kg^-1 BW GABA feeding extremely significantly increased the daily gain of piglets. There was no significant difference in the diarrhea rate of the control group, OS group and HD+OS group in the first, middle and late stages (P >0.05). The MDA level of hippocampus in the OS group was significantly higher than that of the control group and HD+OS group (P <0.01), the T-AOC and GSH levels were extremely significantly lower than the other two groups (P<0.01), which showed that GABA feeding would improve the antioxidant capacity of hippocampus. The levels of GABA_Aand GABA_B receptors in the hippocampus of the HD+OS group were extremely significantly higher than those in the control group and the OS group (P <0.01), which showed that GABA feeding increased the levels of GABA_A and GABA_B in hippocampus. The Bcl-2 level in the hippocampus of the OS group was significantly lower than that of the control group (P <0.05), and the Bax and Caspase-3 levels were extremely significantly higher than those of the control group (P <0.01). The Bcl-2 level of the HD+OS group was extremely significantly higher than that of the OS group (P <0.01), the Bax level was extremely significantly lower than that of the OS group (P <0.01), and the Caspase-3 level was significantly lower than that of the OS group (P < 0.05). Consistent with this, the Bax and Caspase-3 levels in hippocampal neurons of rats in the OS group, GABA+OS+Picrotoxin group and GABA+OS+CGP54626 group were significantly higher than those in the control group and GABA group (P <0.05), which indicated that GABA alleviated the damage of hippocampal neurons under oxidative stress, and the addition of GABA receptor inhibitors blocked the anti-stress damage effect of GABA. 【Conclusion】GABA reduced the level of oxidative stress in the hippocampus of piglets. The anti-stress mechanism of GABA could be related to the decrease of the expression of apoptotic protein gene, while GABA_A and GABA_B receptors mediated this process.

Key words: GABA, piglet, oxidative stress, receptor, apoptosis

杨昌沛, 王乃秀, 汪锴, 黄子晴, 林海烂, 张莉, 张晨, 冯露秋, 甘玲. 最新录用：外源性γ-氨基丁酸对仔猪抗氧化应激作用的效果及机制[J]. 中国农业科学, 0, (): 1-.

YANG ChangPei, WANG NaiXiu, WANG Kai, HUANG ZiQing, LIN HaiLan, ZHANG Li, ZHANG Chen , FENG LuQiu , GAN Lin , .

Effects and Mechanisms of Exogenous GABA against Oxidative Stress in Piglets [J]. Scientia Agricultura Sinica, 0, (): 1-.

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最新录用：外源性γ-氨基丁酸对仔猪抗氧化应激作用的效果及机制

Effects and Mechanisms of Exogenous GABA against Oxidative Stress in Piglets

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