杨宗英^2,3,
杨移斌⁴,
张一柳²,
胡鲲²,
曾柳根³,
刘力硕²,
闫子君²,
杨先乐^2,#,
常藕琴¹
1. 农业部渔用药物创制重点实验室, 广东省水产动物免疫技术重点实验室, 中国水产科学研究院珠江水产研究所, 广州 510380;
2. 上海海洋大学国家水生动物病原库, 上海 201306;
3. 南昌市农业科学院, 南昌 330038;
4. 中国水产科学研究院长江水产研究所, 武汉 430223

作者简介: 杨宗英(1988-),女,博士研究生,研究方向为水产生物免疫学与病害控制,E-mail:lxyangzongying@163.com.

基金项目: 农业部渔用药物创制重点实验室开放课题资助


中图分类号: X171.5

Effects of Avermectin Exposure on Oxidative Stress and Histological Structure of Hepatopancreas in Eriocheir sinensis

Yang Zongying^2,3,
Yang Yibin⁴,
Zhang Yiliu²,
Hu Kun²,
Zeng Liugen³,
Liu Lishuo²,
Yan Zijun²,
Yang Xianle^2,#,
Chang Ouqin¹
1. Key Laboratory of Fishery Drug Development, Ministry of Agriculture;Key Laboratory of Aquatic Animal Immune Technology, Guangdong Province;Pearl River Fishery Research Institute, Chinese Academy of Fishery Sciences, Guangzhou 510380, China;
2. National Pathogen Collection Center for Aquatic Animals, Shanghai Ocean University, Shanghai 201306, China;
3. Nanchang Academy of Agricultural Sciences, Nanchang 330038, China;
4. Yangtze River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Wuhan 430223, China

CLC number: X171.5

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摘要:采用半静态染毒法，研究不同浓度(1.6、8.0、16.0 μg·L^-1)阿维菌素在胁迫中华绒螯蟹12、24、48、96 h时肝胰腺超氧化物歧化酶(SOD)、过氧化氢酶(CAT)活性和丙二醛(MDA)积累量等氧化胁迫相关指标的变化；并在胁迫60 d时，记录各实验组和对照组中华绒螯蟹肝胰腺颜色的变化并进行组织病理观察。3个阿维菌素浓度组为实验组，空白组和溶剂组为对照组。结果表明在整个实验过程中空白组和溶剂组各个时间点各项指标之间无显著差异(P > 0.05)；低浓度(1.6 μg·L^-1)组SOD和CAT活力变化不显著(P > 0.05)；中浓度(8.0 μg·L^-1)组SOD和CAT活力变化趋势一致，持续被诱导，SOD活力在24 h以后显著高于空白组(P<0.01)，CAT活力在48 h以后显著高于空白组(P<0.01)；高浓度(16.0 μg·L^-1)组SOD和CAT活力在12 h~24 h之间变化趋势一致，表现为显著升高(P<0.05)，但是在48 h之后，SOD活力显著下降(P<0.05)；96 h时，高浓度组SOD活力显著低于空白组(P<0.01)，而CAT活力显著高于空白组(P<0.01)；在整个实验过程中各实验组MDA积累量逐渐增加，高浓度组氧化胁迫指标的变化幅度大于低浓度组。胁迫60 d时，取样观察可见对照组中华绒螯蟹肝胰腺正常，各实验组中华绒螯蟹肝胰腺发生了不同程度的病变，肝胰腺颜色由最开始的橘黄色变成了淡黄色、黄白色甚至白色，高、中、低各浓度组肝胰腺白化所占的比例分别是66.6%、57.1%和25.0%。组织病理分析显示，淡黄色肝胰腺的病理变化主要是B细胞数量减少，肝胰腺上皮细胞部分肿大并出现空泡；黄白肝病理观察可见肝胰腺上皮细胞中空泡数量增多，体积增大，且空泡内出现内容物，细胞核固缩；白肝的病理表现主要是基膜增厚，上皮细胞大量脱落，散落在管腔中，空泡数量进一步增多，肝胰腺的生理功能受损。由此得出，阿维菌素会对中华绒螯蟹肝胰腺造成氧化胁迫和组织损伤，且阿维菌素和对肝胰腺的氧化胁迫效应和组织结构损伤程度有一定的剂量-效应关系。
关键词: 阿维菌素/
中华绒螯蟹/
氧化胁迫指标/
肝胰腺组织结构

Abstract:An acute semi-static toxic test was carried out to study the modulation of antioxidant enzymes such as superoxide dismutase (SOD), catalase (CAT) and malondialdehyde content (MDA), which was the main product of lipid peroxidation, at 12, 24, 48 and 96 h in hepatopancreas when Eriocheir sinensis was exposed to different avermectin concentrations including 1.6, 8.0 and 16.0 μg·L^-1. Histological structure and color variations of hepatopancreas were observed on day 60 in test group, blank group and vehicle group, respectively. Three different avermectin concentration groups were test groups, and blank group and vehicle group were control groups. Results were as follows: (1) The three indicators had no significant differences at different time in both vehicle group and blank group, and vehicle group also had no obvious differences from blank group. SOD and CAT activities at the concentration of 1.6 μg·L^-1 also had no significant differences from control groups (P > 0.05). The activities of SOD and CAT showed the similar variation in 8.0 μg·L^-1 group, and they were induced continuously throughout the experiment. SOD activity increased extremely significantly compared with blank group (P<0.01) after 24 h, and CAT activity was extremely significantly higher than blank group after 48 h (P<0.01). SOD and CAT activities were all induced between 12 h and 24 h at the concentration of 16.0 μg·L^-1, but SOD activity decreased significantly after 48 h (P<0.05). At 96 h, the activity of SOD was extremely lower than blank group. However, CAT activity was still extremely significantly higher than blank group (P<0.01). MDA levels of test groups gradually increased along with the increase of stress time of avermectin, and the oxidative stress indicators of low avermectin concentration group varied less than that of high concentration group. (2) Histological structure and color of hepatopancreas were affected at different degrees at different avermectin concentrations for 60 days. The hepatopancreas of E. sinensis of control groups was normal, but hepatopancreas of test groups changed from orange red to light yellow, yellowish-white or stark white. The percentage of white hepatopancreas of 16.0 μg·L^-1, 8.0 μg·L^-1 and 1.6 μg·L^-1 groups was 66.6%, 57.1% and 25.0%, respectively. Histopathological analysis revealed the different pathological lesions of hepatopancreas of different colors. The number of B cells of hepatopancreas was decreased, and some hepatic cells were swelling and had cavity in hepatopancreas of light yellow. When the color of hepatopancreas changed to yellowish-white, the degree of deterioration of hepatopancreas aggravated. The number and volume of vacuoles increased, some contents were present in the vacuoles, and nucleus were pyknotic. In the white hepatopancreas, the basement membrane became thicker, some hepatic cells were crushing and many cell debris appeared in the lumen, many vacuoles existed between the basement membrane and epithelial cells, and the physiological function of hepatopancreas was impaired. The results indicated that avermectin could result in oxidative stress and tissue damage in hepatopancreas of E. sinensis, and the oxidative stress effects and the degree of pathological lesions had a dose-effect relationship with avermectin.
Key words:avermectin/
Eriocheir sinensis/
oxidative stress indicators/
hepatopancreas structure.