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阿维菌素对中华绒螯蟹肝胰腺细胞的细胞毒性:一项体外试验

Cytotoxicity induced by abamectin in hepatopancreas cells of Chinese mitten crab, Eriocheir sinensis: An in vitro assay.

作者信息

Hong Yuhang, Huang Yi, Dong Yanzhen, Xu Dayong, Huang Qiang, Huang Zhiqiu

机构信息

Key Laboratory of Application of Ecology and Environmental Protection in Plateau Wetland of Sichuan, Xichang University, Xichang 415000, Sichuan, China; Key Laboratory of Animal Disease Detection and Prevention in Panxi District, Xichang University, Xichang 415000, China.

Key Laboratory of Application of Ecology and Environmental Protection in Plateau Wetland of Sichuan, Xichang University, Xichang 415000, Sichuan, China.

出版信息

Ecotoxicol Environ Saf. 2023 Jun 28;262:115198. doi: 10.1016/j.ecoenv.2023.115198.

Abstract

Toxic effects of abamectin on non-target aquatic organisms have been well documented due to its extensive use in both agricultural and aquacultural areas. However, knowledge of the abamectin induced cytotoxicity in crustacean hepatopancreas is still incomplete. In this study, we investigated the cytotoxic effects of abamectin on hepatopancreas cells of Chinese mitten crab, Eriocheir sinensis by an in vitro assay. The results showed that abamectin inhibited cell viability with elevated reactive oxygen species (ROS) and malondialdehyde (MDA) levels in a dose-dependent manner. Increased olive tail moment (OTM) values and 8-hydroxy-2'-deoxyguanosine (8-OHdG) contents indicate the DNA damage under abamectin exposure. The up-regulation of the typical apoptosis-related protein BCL2-associated X protein (Bax) and the down-regulation of B cell leukemia/lymphoma 2 (Bcl-2) demonstrate apoptosis in hepatopancreas cells. Meanwhile, the activities of both caspase-3 and caspase-9 were increased, indicating caspase-mediated apoptosis. In addition, qRT-PCR results showed the up-regulation of antioxidant genes superoxide dismutase (SOD) and catalase (CAT). The mRNA expression of Cap 'n' Collar isoform-C (CncC) and c-Jun NH2-terminal kinases (JNK) was also significantly increased, implying the involvement of the Nrf2/MAPK pathway in the antioxidative response. The alteration of innate immune-associated genes Toll-like receptor (TLR) and myeloid differentiation primary response gene 88 (Myd88) also indicates the influence of abamectin on immune status. In summary, the present study reveals the cytotoxicity of abamectin on hepatopancreas cells of E. sinensis and this in vitro cell culture model could be used for further assessment of pesticide toxicity.

摘要

由于阿维菌素在农业和水产养殖领域的广泛使用,其对非靶标水生生物的毒性作用已有充分记录。然而,关于阿维菌素诱导甲壳类动物肝胰腺细胞毒性的知识仍不完整。在本研究中,我们通过体外试验研究了阿维菌素对中华绒螯蟹肝胰腺细胞的细胞毒性作用。结果表明,阿维菌素以剂量依赖的方式抑制细胞活力,同时活性氧(ROS)和丙二醛(MDA)水平升高。橄榄尾矩(OTM)值和8-羟基-2'-脱氧鸟苷(8-OHdG)含量的增加表明阿维菌素暴露下的DNA损伤。典型凋亡相关蛋白Bax的上调和B细胞淋巴瘤/白血病-2(Bcl-2)的下调表明肝胰腺细胞发生凋亡。同时,caspase-3和caspase-9的活性均增加,表明caspase介导的凋亡。此外,qRT-PCR结果显示抗氧化基因超氧化物歧化酶(SOD)和过氧化氢酶(CAT)上调。Cap 'n' Collar异构体C(CncC)和c-Jun氨基末端激酶(JNK)的mRNA表达也显著增加,这意味着Nrf2/MAPK途径参与了抗氧化反应。先天免疫相关基因Toll样受体(TLR)和髓样分化初级反应基因88(Myd88)的改变也表明阿维菌素对免疫状态的影响。总之,本研究揭示了阿维菌素对中华绒螯蟹肝胰腺细胞的细胞毒性,这种体外细胞培养模型可用于进一步评估农药毒性。

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