中国农业科学 ›› 2022, Vol. 55 ›› Issue (17): 3437-3449.doi: 10.3864/j.issn.0578-1752.2022.17.014
杨昌沛1(),王乃秀1,汪锴1,黄子晴1,林海烂1,张莉1,张晨1,冯露秋1,甘玲1,2,*()
收稿日期:
2021-07-25
接受日期:
2022-06-16
出版日期:
2022-09-01
发布日期:
2022-09-07
通讯作者:
甘玲
作者简介:
杨昌沛,E-mail: 基金资助:
YANG ChangPei1(),WANG NaiXiu1,WANG Kai1,HUANG ZiQing1,LIN HaiLan1,ZHANG Li1,ZHANG Chen1,FENG LuQiu1,GAN Ling1,2,*()
Received:
2021-07-25
Accepted:
2022-06-16
Online:
2022-09-01
Published:
2022-09-07
Contact:
Ling GAN
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(20 mg·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GABAA和GABAB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GABAA和GABAB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GABAA和GABAB受体介导了该过程。
杨昌沛,王乃秀,汪锴,黄子晴,林海烂,张莉,张晨,冯露秋,甘玲. 外源性γ-氨基丁酸抵抗仔猪氧化应激的效果及机制[J]. 中国农业科学, 2022, 55(17): 3437-3449.
YANG ChangPei,WANG NaiXiu,WANG Kai,HUANG ZiQing,LIN HaiLan,ZHANG Li,ZHANG Chen,FENG LuQiu,GAN Ling. Effects and Mechanisms of Exogenous GABA Against Oxidative Stress in Piglets[J]. Scientia Agricultura Sinica, 2022, 55(17): 3437-3449.
表1
不同浓度GABA对仔猪血清氧化/抗氧化指标和ACTH水平的影响"
项目 Items | 组别 Groups | P值 P value | ||||
---|---|---|---|---|---|---|
Con | OS | HD+OS | MD+OS | LD+OS | ||
丙二醛 Malondialdehyde (nmol·mL-1) | 6.56±0.95 | 33.88±1.97## | 4.98±0.5** | 13.43±0.86** | 16.42±0.86** | <0.001 |
谷胱甘肽 Glutathione (μmol·L-1) | 19.20±0.4 | 12.56±1.26## | 37.15±1.14** | 32.71±0.21** | 28.89±0.24** | <0.001 |
总抗氧化能力 Total antioxidant capacity (U·mL-1) | 2.82±0.06 | 2.19±0.13## | 4.26±0.07** | — | — | <0.001 |
促肾上腺皮质激素 Adreno-cortico-tropic-hormone (ng·L-1) | 70.85±7.22 | 100.09±4.67## | 72.91±2.91* | — | — | 0.01 |
表3
高浓度GABA对仔猪海马组织氧化/抗氧化相关指标的影响(平均值±标准误,n=5)"
项目 Items | 组别 Groups | P值 P value | ||
---|---|---|---|---|
Con | OS | HD+OS | ||
丙二醛 Malondialdehyde (nmol·mL-1) | 137.4±1.43 | 430.27±2.33## | 148.06±4.07** | <0.001 |
谷胱甘肽 Glutathione (μmol·L-1) | 34.3±2.11 | 14.72±1.4## | 29.91±2.08** | <0.001 |
总抗氧化能力 Total antioxidant capacity (U·mL-1) | 1.94±0.07 | 0.19±0.05## | 2.87±0.23** | <0.001 |
表5
GABA受体抑制剂对大鼠海马神经元GABA受体含量的影响(平均值±标准误,n=3)"
项目 Items | 组别 Groups | P值 P value | ||||
---|---|---|---|---|---|---|
Con | OS | GABA+OS | GABA+OS+ Picrotoxin | GABA+OS+ CGP54626 | ||
GABAA型受体 GABAA receptor (μmol·L-1) | 12.25±0.16 | 13.18±0.46 | 13.85±0.15## | 9.97±0.50## | 13.93±0.22## | <0.001 |
GABAB型受体 GABAB receptor (μmol·L-1) | 7.18±0.43 | 8.50±0.19 | 9.67±0.54## | 9.56±0.62## | 6.92±0.37 | 0.004 |
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