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急性缺氧对大弹涂鱼(Boleophthalmus pectinirostris)肝脏中缺氧应答相关基因表达、蛋白含量及酶活性的影响
任倩妍1, 张木子2, 黎明2, 王日昕2, 石戈1
1.浙江海洋大学海洋科学与技术学院 舟山 316022;2.宁波大学海洋学院 宁波 315211
摘要:
为了评估缺氧对大弹涂鱼的影响,本研究通过模拟缺氧环境(溶解氧1.5±0.11mg/L),探究急性缺氧(1h,3h和6h)对大弹涂鱼缺氧相关基因表达、蛋白含量及酶活性的影响。结果显示,6h缺氧过程中,对照组实验鱼肝脏中缺氧相关基因表达、蛋白含量和酶活性无显著性差异;缺氧处理组实验鱼肝脏中缺氧诱导因子(HIF)、葡萄糖转运体、脯氨酰羟化酶和Bcl2/腺癌E1B19KD交互蛋白3基因的表达量和蛋白含量在6h处理时间内逐渐上调,但肿瘤抑制蛋白p53基因的表达量和蛋白含量在1h时显著最高;缺氧处理组实验鱼肝脏中乳酸脱氢酶、柠檬酸合酶和丙酮酸激酶活性逐渐下调,在6h时达到最低;但肝脏中葡萄糖激酶和磷酸果糖激酶活性在3h时显著增加,随后逐渐降低,在6h时达到最低;在整个缺氧过程中,缺氧处理组实验鱼肝脏中缺氧相关基因表达、蛋白含量和酶活性显著高于对照组。本研究结果表明,HIF信号通路在缺氧应答中被激活;缺氧胁迫会抑制鱼类需氧代谢的生理过程。
关键词:  大弹涂鱼  缺氧  基因表达  蛋白含量  酶活性
DOI:10.11693/hyhz20171100301
分类号:Q789;S941
基金项目:国家自然科学基金项目,31472279号,31502176号;宁波市自然科学基金项目,2016A610083号。
DIFFERENTIAL INDUCTION OF GENE EXPRESSIONS, PROTEIN CONTENTS AND ENZYME ACTIVITIES INVOLVED IN HYPOXIC RESPONSIVE IN LIVER TISSUES OF MUDSKIPPER BOLEOPHTHALMUS PECTINIROSTRIS EXPOSED TO ACUTE HYPOXIA
REN Qian-Yan1, ZHANG Mu-Zi2, LI Ming2, WNAG Ri-Xin2, SHI Ge1
1.School of Marine Science and Technology, Zhejiang Ocean University, Zhoushan 316022, China;2.School of Marine Science, Ningbo University, Ningbo 315211, China
Abstract:
To evaluate the effect of hypoxia on mudskipper, a study was carried out to simulated acute hypoxia (dissolved oxygen:1.5±0.11mg/L) in the laboratory and analyzed gene expressions, protein contents and enzyme activities in mudskipper exposed to various durations of acute hypoxia (1h, 3h and 6h). No significant differences were found in gene expressions, protein contents and enzyme activities in control group. Fish exposed to acute hypoxia, liver mRNA expression and protein content of hypoxia-inducible factor 1, glucose transporters, prolyl-4-hydroxylase and Bcl-2/adenocarcinoma E1B19KD interacting protein 3 gradually increased at hour 6, on the contrary, fish exposed to hypoxia at hour 1 had the highest mRNA expression and protein content of tumour suppressor protein p53. Liver lactate dehydrogenase, citroyl synthetase and pyruvate kinase activities gradually decreased from hour 1 to hour 6, the lowest significant values were found at hour 6. However, liver glucokinase and phosphofructokinase activities abruptly increased at hour 3, and then gradually decreasing, the lowest values were found at hour 6. During the 6-hour period, gene expressions, protein contents and enzyme activities involved in hypoxic responsive of fish in the hypoxia treatment group were higher than those of fish in control group. This study indicates that the HIF signaling pathway is activated in response to hypoxia; liver enzymes are decreased, which indicated a depression in the capacity of aerobic metabolism during fish hypoxia.
Key words:  Boleophthalmus pectinirostris  hypoxic  gene expression  protein content  enzyme activity
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