文献类型： 外文期刊

作者： Wang, Jingqian ¹ ; Gao, Xinming ¹ ; Wang, Li ¹ ; Du, Chen ¹ ; Hou, Congcong ¹ ; Liu, Feng ² ; Xie, Qingping ² ; Lou, Bao ² ; Jin, Shan ¹ ; Zhu, Junquan ¹ ;

作者机构： 1.Ningbo Univ, Sch Marine Sci, Key Lab Appl Marine Biotechnol, Minist Educ, Ningbo 315211, Zhejiang, Peoples R China

2.Zhejiang Acad Agr Sci, Hangzhou 310021, Zhejiang, Peoples R China

关键词： Apoptosis; Cell line; Hypoxic stress; Larimichthys polyactis; ROS

期刊名称：AQUACULTURE （ 影响因子：4.5； 五年影响因子：4.6 ）

ISSN： 0044-8486

年卷期： 2023 年 563 卷

页码：

收录情况： SCI

摘要： Cell lines play an important role in studying biological processes in vitro. In this study, we aimed to establish and characterize the first cell line isolated from the small yellow croaker (Larimichthys polyactis), an economically important marine fish in China. The small yellow croaker fry (SYCF) cells proliferated well in Leibovitz's 15 medium containing 2% antibiotics, 0.1 M NaCl, and 10% fetal bovine serum at 27 degrees C. Sequence analyses of mitochondrial genes confirmed the origin of SYCF cells. Furthermore, we unraveled the mechanism of hypoxia-induced apoptosis, a major threat to aquatic animals using the SYCF cell line. The HIF-PHD pathway of the SYCF cells can be activated under hypoxic stress, suggesting that this cell line is impressible for hypoxia and can serve as a superb model in vitro to study the cell physiology in L. polyactis in a hypoxic setting. We found that hypoxia results in morphological abnormalities, excess reactive oxygen species (ROS) production, and dysfunctional redox homeostasis, indicating the occurrence of oxidative stress in the SYCF cells. In addition, decreased cell viability and severe apoptosis were detected under hypoxic conditions. Through monitoring the indicators of the intrinsic and extrinsic apoptosis pathways, including the mitochondrial membrane potential, the expression of bax, bcl2, bnip3, and fas, Bcl2, and Caspase-3 protein, and activity of Caspases-3/8/9, we found that both pathways are involved. Considering that ROS-induced oxidative stress may be the main reason of apoptosis, we investigated whether the addition of an antioxidant, N-Acetyl-L-cysteine (NAC), will ameliorate the apoptosis. Interestingly, we found that both intrinsic and extrinsic apoptosis indicators were markedly recovered by adding NAC in culture media, significant remissions were also observed in the cell viability and apoptosis rate. These results suggested that hypoxia-induced apoptosis of SYCF cells may be mediated by the overproduction of ROS, and the antioxidant may be a useful material to relieve cell damage under hypoxia in fish. In general, this study established a marine fish cell line of SYCF cells, which can be used as an in vitro research model of L. polyactis and for revealing the mechanisms of ROS-induced apoptosis under hypoxic conditions in this species. This provides important insights into the cellular physiology of marine fish under hypoxia.