| 急性高温胁迫对翘嘴鳜幼鱼抗氧化酶和消化酶活性及热休克蛋白基因表达的影响 |
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| 引用本文： | 张晨光,丁炜东,曹哲明,邴旭文,徐畅,李良.急性高温胁迫对翘嘴鳜幼鱼抗氧化酶和消化酶活性及热休克蛋白基因表达的影响[J].南方农业学报,2021,52(3):815-826. |
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| 作者姓名： | 张晨光 丁炜东 曹哲明 邴旭文 徐畅 李良 |
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| 作者单位： | 南京农业大学无锡渔业学院,江苏无锡 214081;中国水产科学研究院淡水渔业研究中心,江苏无锡 214801;南京农业大学无锡渔业学院,江苏无锡 214081;中国水产科学研究院淡水渔业研究中心,江苏无锡 214801 |
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| 基金项目： | 江苏省农业自主创新基金项目[CX（17）3005]；中国水产科学研究院淡水渔业研究中心基本科研业务费专项（2019JBFZ01） |
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| 摘 要： | 【目的】从抗氧化酶和消化酶活性及热休克蛋白基因表达层面明确高温胁迫对翘嘴鳜（Siniperca chuatsi）幼鱼生长及应激生理响应的影响，为实际生产中翘嘴鳜幼鱼培育提供可靠的参考依据。【方法】对2月龄翘嘴鳜幼鱼进行96 h的急性高温胁迫，通过预试验测试高起始致死温度（96 h-UILT50），采用突变升温方法设常温对照组（26.0℃）和急性高温胁迫组（36.0℃），分别于胁迫0、6、12、24、48和96 h后取样，使用生化试剂盒测定抗氧化酶和消化酶活性，并以实时荧光定量PCR检测热休克蛋白基因（HSP70α和HSP90α）的表达情况。【结果】翘嘴鳜幼鱼死亡率随水温的升高不断上升，其96 h-UILT50为36.22℃。在96 h的急性高温胁迫过程中，翘嘴鳜幼鱼肝脏超氧化物歧化酶（SOD）和过氧化氢酶（CAT）活性呈降低-升高-降低的变化趋势，谷丙转氨酶（GPT）活性及丙二醛（MDA）含量则表现出升高-降低-升高的变化趋势；在消化酶方面，翘嘴鳜幼鱼胃蛋白酶和肠道淀粉酶（AMS）活性呈降低-升高-降低的变化趋势，而肠道脂肪酶（LPS）活性呈升高-降低-升高的变化趋势。在急性高温胁迫过程中，翘嘴鳜幼鱼HSP70α基因相对表达量呈升高-下降的波动式变化趋势，于胁迫12 h时达最高值，在胁迫48 h时出现第2个峰值；HSP90α基因表达呈先升高后降低的变化趋势，于胁迫24 h时上调至最高值；但至胁迫96 h时HSP70α和HSP90α基因的相对表达量仍显著高于对照组翘嘴鳜幼鱼（P<0.05）。【结论】急性高温胁迫对翘嘴鳜幼鱼抗氧化酶和消化酶活性及热休克蛋白基因表达产生显著影响。其中，热休克蛋白基因HSP70α和HSP90α参与高温胁迫应答过程的生理调节，以应对高温胁迫对肝脏细胞的损伤，故可作为高温胁迫应答的标志物。
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| 关 键 词： | 翘嘴鳜 高温胁迫 抗氧化酶 消化酶 热休克蛋白基因 |
| 收稿时间： | 2020-07-22 |
Effects of acute high temperature stress on antioxidant enzymes activity,digestive enzymes activity and gene expression of heat shock proteins in mandarin fish(Siniperca chuatsi) |
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| ZHANG Chen-guang,DING Wei-dong,CAO Zhe-ming,BING Xu-wen,XU Chang,LI Liang.Effects of acute high temperature stress on antioxidant enzymes activity,digestive enzymes activity and gene expression of heat shock proteins in mandarin fish(Siniperca chuatsi)[J].Journal of Southern Agriculture,2021,52(3):815-826. |
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| Authors: | ZHANG Chen-guang DING Wei-dong CAO Zhe-ming BING Xu-wen XU Chang LI Liang |
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| Institution: | 1. Wuxi Fisheries College, Nanjing Agricultural University, Wuxi, Jiangsu 214081, China;2. Fisheries Research Center, Chinese Academy of Fishery Sciences, Wuxi, Jiangsu 214081, China |
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| Abstract: | 【Objective】The effects of high temperature stress on the growth and physiological response of juvenile mandarin fish(Siniperca chuatsi) were investigated from the aspects of antioxidant enzyme, digestive enzyme activities and heat shock protein family gene expression, which provided a reliable reference for the cultivation of juvenile S. chuatsi in practical production.【Method】Two-month-old juvenile S. chuatsi was subjected to acute high temperature stress for 96 h. The high initial lethal temperature(96 h-UILT50) was measured by pre-test. The control group(26.0℃) and the acute high temperature stress group(36.0℃) were set by abrupt temperature rise method. Samples were taken at 0, 6, 12, 24, 48 and 96 h respectively after stress. Biochemistry kits were used to determine the activities of antioxidant and digestive enzymes, and real-time fluorescence quantitative PCR was used to detect the expression of heat shock protein family genes(HSP70αand HSP90α).【Result】The mortality rate of juvenile S. chuatsi increased with the increase of water temperature, and its 96 h-UILT50 was 36.22℃. During the 96 h acute high temperature stress, the activities of superoxide dismutase(SOD) and catalase(CAT) in liver of S. chuatsi showed a decreasing, increasing and decreasing trend, while the activities of alanine aminotransferase(GPT) and the content of malondialdehyde(MDA) showed an increasing, decreasing and increasing trend. In terms of digestive enzymes, the activities of pepsin and intestinal amylase(AMS) of juvenile S. chuatsi showed a decreasing, increasing and decreasing trend, while the activities of intestinal lipase(LPS) showed an increasing, decreasing and increasing trend. In the process of acute heat stress, the relative expression of HSP70α gene in juvenile S. chuatsi showed a fluctuating trend of increasing to decreasing, and reached the highest value at 12 h of stress, and the second peak value appeared at 48 h of stress. The expression of HSP90α gene increased at first and then decreased, and reached the highest value at 24 h of stress. However, at 96 h of stress, HSP70α and HSP90α genes were still significantly higher than those of juvenile S. chuatsi in the control group(P<0.05).【Conclusion】The activity of antioxidant enzyme, digestive enzyme and the expression of heat shock protein gene of juvenile S. chuatsi are significantly affected by acute high temperature stress. Among them, heat shock protein genes HSP70α and HSP90α are involved in the physiological regulation of the response to high temperature stress to deal with the damage of liver cells under high temperature stress, so they can be used as markers of the response to high temperature stress. |
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