Huang Yongxiong, Li Qi, Yuan Yunhao, Zhang Zhiqiang, Jiang Baijian, Yang Shiping, Jian Jichang
College of Fishery, Guangdong Ocean University, Guangdong Provincial Key Laboratory of Pathogenic Biology and Epidemiology for Aquatic Economic Animal, Key Laboratory of Control for Disease of Aquatic Animals of Guangdong Higher Education Institutes, Southern Marine Science and Engineering Guangdong Laboratory, Zhanjiang, China; Guangdong Provincial Engineering Research Center for Aquatic Animal Health Assessment, Shenzhen, China.
College of Fishery, Guangdong Ocean University, Guangdong Provincial Key Laboratory of Pathogenic Biology and Epidemiology for Aquatic Economic Animal, Key Laboratory of Control for Disease of Aquatic Animals of Guangdong Higher Education Institutes, Southern Marine Science and Engineering Guangdong Laboratory, Zhanjiang, China.
Fish Shellfish Immunol. 2022 Mar;122:257-267. doi: 10.1016/j.fsi.2022.02.010. Epub 2022 Feb 8.
Nuclear factor E2-related factor 2 (Nrf2) is a multifunctional transcription factor that plays an important role in antioxidant activities. However, its effect on antioxidant capacity in Litopenaeus vannamei, an economically important crustacean, remains unclear. In this study, the role of Nrf2 in response to oxidative stress in L. vannamei was determined by its effect on relevant gene expression and enzymatic activity. Nrf2 was cloned and analyzed. Results revealed that Nrf2 contains a 1575 bp open reading frame encoding 524 amino acids and a conserved bZIP Maf domain. The sequence similarity of Nrf2 between L. vannamei and Homarus americanus is 81%. Although the Nrf2 expression was detected in all tissues, the Nrf2 expression levels were the highest in the hepatopancreas, followed by the eyestalk and muscle. RNA interference significantly decreased the expression of antioxidant-related genes (SOD, GPX, CAT, Trx, and HO-1; p < 0.05), significantly upregulated the expression of autophagy genes (Atg3, Atg4, Atg5, Atg10, and Atg12; p < 0.05) and apoptosis genes (Caspase-3 and P53; p < 0.05). Moreover, SOD, CAT, and GPX enzyme activities decreased whereas the MDA activity increased. The histological results of the shrimp injected with dsRNA-Nrf2 showed that the hepatic tubules were irregularly arranged, the lumen was abnormal, and a few hepatic tubules were significantly enlarged compared with those of the dsRNA-EGFP group. The hepatocytes were also vacuolated. In conclusion, this study provided evidence that Nrf2 is involved in the regulation of antioxidant capacity, oxidative stress, apoptosis, and autophagy in shrimp.
核因子E2相关因子2（Nrf2）是一种多功能转录因子，在抗氧化活性中发挥重要作用。然而，其对经济上重要的甲壳类动物凡纳滨对虾抗氧化能力的影响仍不清楚。在本研究中，通过Nrf2对相关基因表达和酶活性的影响来确定其在凡纳滨对虾应对氧化应激中的作用。对Nrf2进行了克隆和分析。结果显示，Nrf2包含一个1575 bp的开放阅读框，编码524个氨基酸以及一个保守的bZIP Maf结构域。凡纳滨对虾和美洲螯龙虾之间Nrf2的序列相似性为81%。虽然在所有组织中均检测到Nrf2表达，但Nrf2在肝胰腺中的表达水平最高，其次是眼柄和肌肉。RNA干扰显著降低了抗氧化相关基因（超氧化物歧化酶、谷胱甘肽过氧化物酶、过氧化氢酶、硫氧还蛋白和血红素加氧酶-1；p<0.05）的表达，显著上调了自噬基因（自噬相关基因3、自噬相关基因4、自噬相关基因5、自噬相关基因10和自噬相关基因12；p<0.05）和凋亡基因（半胱天冬酶-3和P53；p<0.05）的表达。此外，超氧化物歧化酶、过氧化氢酶和谷胱甘肽过氧化物酶的酶活性降低，而丙二醛活性增加。注射dsRNA-Nrf2的对虾的组织学结果显示，与dsRNA-绿色荧光蛋白组相比，肝小管排列不规则，管腔异常，少数肝小管明显增大。肝细胞也出现空泡化。总之，本研究提供了证据表明Nrf2参与了对虾抗氧化能力、氧化应激、凋亡和自噬的调节。
