Zhou Xiujuan, Chen Qihui, Chen Luna, Liao Xinqi, Wang Zuoyuan, Zhu Fei
Key Laboratory of Applied Technology on Green-Eco-Healthy Animal Husbandry of Zhejiang Province, College of Animal Science and Technology & College of Veterinary Medicine, Zhejiang Agriculture and Forestry University, Hangzhou, 311300, China.
Key Laboratory of Applied Technology on Green-Eco-Healthy Animal Husbandry of Zhejiang Province, College of Animal Science and Technology & College of Veterinary Medicine, Zhejiang Agriculture and Forestry University, Hangzhou, 311300, China.
Fish Shellfish Immunol. 2023 Oct;141:109075. doi: 10.1016/j.fsi.2023.109075. Epub 2023 Sep 18.
Reactive oxygen species (ROS) are typically regarded as being generated by the cellular respiratory chain or by cells under pathological damage, which play a crucial role as signaling molecules in promoting hemocytes circulation and normal cellular physiological functions. In this study, the antioxidant N-acetylcysteine (NAC) was used to reduce ROS in vivo and in vitro, which to analyze the effect of ROS on innate immunity and viral infection of mud crab. The total hemocyte count (THC), phenoloxidase (PO), superoxide dismutase (SOD) activity, immune-relative genes were analyzed, respectively. Moreover, the effect of ROS on WSSV infection was analyzed by THC and hemocytes apoptosis. The data showed that NAC could effectively remove and inhibit intracellular ROS. The THC of NAC group was reduced at 12 h and 24 h compared with that of control. And the inhibition of ROS by NAC could increase the SOD activity with control group, while increased the PO activity caused by early WSSV infection. And NAC could up-regulate the expression of MCM7, JAK, TLR and proPO significantly, while down-regulate the expression of Astakine, proPO, caspase and p53. Similarly, NAC could inhibit WSSV-induced apoptosis of S. paramamosain hemocytes. The data illustrated that ROS participates in the interaction between hemocytes and virus infection by regulating innate immunity. Especially, after NAC inhibited ROS, the expression of hemocytes proliferation gene Astakine was also inhibited, which may indicate that ROS is related to the process of hemocytes proliferation. The data will show a preliminary exploration on the regulatory role of ROS in crustacean immune system.
活性氧（ROS）通常被认为是由细胞呼吸链或处于病理损伤状态的细胞产生的，它们作为信号分子在促进血细胞循环和正常细胞生理功能方面发挥着关键作用。在本研究中，抗氧化剂N - 乙酰半胱氨酸（NAC）被用于在体内和体外降低ROS，以分析ROS对青蟹先天免疫和病毒感染的影响。分别分析了总血细胞计数（THC）、酚氧化酶（PO）、超氧化物歧化酶（SOD）活性以及免疫相关基因。此外，通过THC和血细胞凋亡分析了ROS对白斑综合征病毒（WSSV）感染的影响。数据表明，NAC可以有效清除和抑制细胞内ROS。与对照组相比，NAC组在12小时和24小时时THC降低。NAC对ROS的抑制作用可使SOD活性与对照组相比增加，同时增加早期WSSV感染引起的PO活性。并且NAC可以显著上调MCM7、JAK、TLR和前酚氧化酶原（proPO）的表达，同时下调Astakine、proPO、半胱天冬酶（caspase）和p53的表达。同样，NAC可以抑制WSSV诱导的拟穴青蟹血细胞凋亡。数据表明，ROS通过调节先天免疫参与血细胞与病毒感染之间的相互作用。特别是，NAC抑制ROS后，血细胞增殖基因Astakine的表达也受到抑制，这可能表明ROS与血细胞增殖过程有关。这些数据将展示对ROS在甲壳类免疫系统中调节作用的初步探索。
