pH和菲协同效应对中华绒螯蟹幼蟹的毒性效应

杨志刚; 江青青; 房余程; 陈阿琴; 成永旭; 王爱民

doi:10.12131/20230068

pH和菲协同效应对中华绒螯蟹幼蟹的毒性效应

doi: 10.12131/20230068

杨志刚^{1, 2, 3,},
江青青^{1, 2, 3},
房余程^{1, 2, 3},
陈阿琴^{1, 2, 3, ,},
成永旭^{1, 2, 3},
王爱民⁴

1.
上海海洋大学/农业农村部淡水水产种质资源重点实验室，上海 201306
2.
上海海洋大学/农业农村部鱼类营养与环境生态研究中心，上海 201306
3.
上海海洋大学/水产科学国家级实验教学示范中心，上海 201306
4.
盐城工学院海洋与生物工程学院，江苏盐城 224000

基金项目: 国家自然科学基金面上项目 (32273154)；上海市自然科学基金项目 (22ZR1427300)；国家现代农业产业技术体系 (CARS-48)；上海市研究生教育学会研究课题 (ShsgeG202215)；黄河三角洲人才工程 (DYRC20190210)

详细信息

作者简介:
杨志刚 (1973—)，男，教授，博士，研究方向为甲壳类动物营养调控及环境毒理学。E-mail: zgyang@shou.edu.cn

通讯作者:
陈阿琴 (1977—)，女，副教授，博士，研究方向为水产动物生殖生理学及应激适应生理学。E-mail: aqchen@shou.edu.cn

中图分类号: S 966.16
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出版历程
- 收稿日期: 2023-04-04
- 修回日期: 2023-06-01
- 录用日期: 2023-06-29
- 网络出版日期: 2023-07-07

Synergistic effects of pH and phenanthrene on toxicity of juvenile Chinese mitten crab (Eriocheir sinensis)

YANG Zhigang^{1, 2, 3
,},
JIANG Qingqing^{1, 2, 3},
FANG Yucheng^{1, 2, 3},
CHEN Aqin^{1, 2, 3
, ,},
CHENG Yongxu^{1, 2, 3},
WANG Aimin⁴

1.
Shanghai Ocean University/Key Laboratory of Freshwater Aquatic Genetic Resources, Ministry of Agriculture and Rural Affairs, Shanghai 201306, China
2.
Shanghai Ocean University/Center for Research on Environmental Ecology and Fish Nutrion (CREEFN) of the Ministry of Agriculture and Rural Affairs, Shanghai 201306, China
3.
Shanghai Ocean University/National Demonstration Center for Experimental Fisheries Science Education, Shanghai 201306, China
4.
College of Marine and Bioengineering, Yancheng Institute of Technology, Yancheng 224000, China

摘要

摘要: 为探究酸化以及环境污染物菲 (Phenanthrene, PHE) 协同效应对中华绒螯蟹 (Eriocheir sinensis) 的影响，开展了不同pH (5.5、6.5、7.8) 和PHE (0、50 µg·L⁻¹) 对中华绒螯蟹 [(15.0±2.3) g] 的联合暴露实验 (为期14 d)。结果发现：1) 中华绒螯蟹在pH 5.5×PHE 50的协同处理下，肝胰腺和鳃组织出现严重病理损伤，肝胰腺小管形状改变，肝管细胞萎缩出现大量空泡，鳃轴肿大，鳃丝角质层表皮破损，局部基膜破裂；而pH 6.5×PHE 50对中华绒螯蟹肝胰腺和鳃组织的损伤并不显著。2) PHE处理引起中华绒螯蟹幼蟹肝胰腺糖原 (Gly) 以及低密度脂蛋白胆固醇 (LDL-C) 浓度显著降低，表明PHE会导致中华绒螯蟹能量代谢水平下降；相较于对照组，pH 5.5×PHE 50对甘油三酯 (TG)、Gly、高密度脂蛋白胆固醇 (HDL-C) 和LDL-C浓度均有抑制效果，而pH 6.5×PHE 50使得中华绒螯蟹TG浓度较对照组显著升高。3) 实时荧光定量PCR结果表明，多环芳烃受体 (ahr)、芳香烃受体核转位因子 (arnt) 和细胞色素p450 1A1 (cyp1a1) 基因的表达量均随pH的降低而升高，并在pH 5.5时达到最高。以上结果表明，酸化和PHE的联合作用改变了中华绒螯蟹的能量代谢，其中pH 5.5时显著加剧了PHE对其肝胰腺组织结构的毒性作用以及对能量代谢的影响。
- 中华绒螯蟹 /
- 酸化 /
- 菲 /
- 能量代谢
Abstract: In order to explore the synergistic effects of acidification and environmental pollutant phenanthrene (PHE) on Chinese mitten crabs (Eriocheir sinensis), we carried out a 14-day joint exposure experiment of E. sinensis [(15.0±2.3) g] to different pH (5.5, 6.5, 7.8) and PHE (0, 50 µg·L⁻¹). The results show that: 1) Under the synergistic treatment of pH 5.5×PHE 50 on E. sinensis, the hepatopancreas and gill tissues were severely damaged; the shape of the hepatopancreatic tubules changed; a large number of vacuoles appeared in the atrophy of the hepatic duct cells. The gill axis was enlarged; the cuticle of the gill filament was damaged; the local basement membrane was ruptured. However, pH 6.5×PHE 50 did not significantly damage the hepatopancreas and gill tissues of E. sinensis. 2) PHE treatment significantly reduced the contents of Gly and LDL-C in hepatopancreas of juvenile E. sinensis, indicating that PHE could decrease the energy metabolism level of E. sinensis. Compared with the control group, pH 5.5×PHE 50 inhibited the contents of TG, Gly, HDL-C and LDL-C, while pH 6.5×PHE 50 significantly increased TG content. 3) Real-time fluorescent quantitative PCR experiment shows that AHR, ARNT and CYP1A1 increased significantly with the decrease of pH, and reached the highest values at pH 5.5. The results reveal that the combining effect of acidification and PHE changes the energy metabolism of E. sinensis, and pH 5.5 significantly aggravates the toxic effect of PHE on the hepatopancreas tissue structure of E. sinensis, as well as its energy metabolism.
- Eriocheir sinensis /
- Acidification /
- Phenanthrene /
- Energy metabolism

HTML全文

图 1 中华绒螯蟹肝胰腺和鳃组织暴露在 PHE (50 µg·L⁻¹) 和 pH (7.8、6.5和5.5) 中14 d 后的病理变化

注：a. pH 7.8×PHE 50组肝胰腺；b. pH 6.5×PHE 50组肝胰腺；c. pH 5.5×PHE 50组肝胰腺；d. pH 7.8×PHE 50组鳃；e. pH 6.5×PHE 50组鳃；f. pH 5.5×PHE 50组鳃。

Figure 1. Pathological changes of hepatopancreas and gill tissues of E. sinensis after exposure to PHE (50 µg·L⁻¹) and pH (7.8, 6.5, and 5.5) for 14 d

Note: a. Hepatopancreas in pH 7.8×PHE 50 group; b. Hepatopancreas in pH 6.5×PHE 50 group; c. Hepatopancreas in pH 5.4×PHE 50 group; d. Gill in pH 7.8×PHE 50 group; e. Gill in pH 6.5×PHE 50 group; f. Gill in pH 5.5×PHE 50 group.

下载: 全尺寸图片幻灯片

图 2 不同 pH 条件下 PHE 浓度对中华绒螯蟹能量代谢指标的影响

注：数值表示为“平均值±标准误”(n=3)；方柱上方不同大写字母表示在PHE 0 µg·L⁻¹下差异显著 (P<0.05)；不同小写字母表示在PHE 50 µg·L⁻¹下差异显著 (P<0.05)；*表示相同pH下组间差异显著 (P<0.05)。图3同此。

Figure 2. Effect of PHE concentration on energy metabolism indexes of E. sinensis under different acidification conditions

Note: Values are represented as Mean±SE (n=3); different uppercase letters above the columns indicate significant differences at PHE 0 µg·L⁻¹ (P<0.05); different lowercase letters indicate significant differences at PHE 50 µg·L⁻¹ (P<0.05); * indicates significant differences between the two groups at the same pH values (P<0.05). The same case in Fig. 3.

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图 3 不同 pH 条件下 PHE 浓度对中华绒螯蟹肝胰腺基因表达量的影响

Figure 3. Effect of different PHE concentrations on expression of hepatopancreas genes in E. sinensis under acidification conditions

下载: 全尺寸图片幻灯片

表 1 引物名称及序列

Table 1. Primer names and sequnences

基因 Gene	对应蛋白 Corresponding protein	引物序列 (5'—3') Primer sequence (5'−3')
ahr	多环芳烃受体 Polycyclic aromatic hydrocarbon receptor	F: GGCGGTAACACCAGTGAAGAGTC R: TGGAGATTGTAGGAGGCGAGAAGT
arnt	芳香烃受体核转位因子 (ARNT) 重组蛋白 Aryl hydrocarbon receptor nuclear translocation factor (ARNT) recombinant protein	F: CCAACCTTCATGCGGCAGATGAAC R: ACACAGGAGCCAGCCAACCAAG
cyp1a1	细胞色素 Cytochrome P1A1	F: ATTTCGTGCTGGTTTGGC R: GGAGTTGCTGCGTATTGGT

下载: 导出CSV

表 2 pH 和 PHE 对中华绒螯蟹肝胰腺甘油三酯、高密度脂蛋白胆固醇、低密度脂蛋白胆固醇和糖原含量影响的双因素方差分析

Table 2. Two-way analysis of variance for influence of pH and PHE on contents of TG, HDL-C, LDL-C and Gly in E. sinensis hepatopancreas

能量代谢指标　　　　 Energy metabolism index	处理组别 Group	自由度 DF	均方差 MS	F	P
甘油三酯 TG	PHE	1	0.009	6.591	<0.001
	pH	2	0.025	17.585	<0.001
	PHE×pH	2	0.018	12.798	<0.001
高密度脂蛋白胆固醇 HDL-C	PHE	1	0.000 2	0.301	0.593
	pH	2	0.002	201.824	<0.001
	PHE×pH	2	0.000 1	18.863	<0.001
低密度脂蛋白胆固醇 LDL-C	PHE	1	0.000 1	9.378	0.010
	pH	2	0.008	145.710	<0.001
	PHE×pH	2	0.001	10.200	0.003
糖原 Gly	PHE	1	0.240	39.171	0.020
	pH	2	0.677	110.327	<0.001
	PHE×pH	2	0.043	6.952	0.010

下载: 导出CSV

表 3 pH 和 PHE 对中华绒螯蟹 ahr、 arnt、 cyp1a1 基因表达量影响的双因素方差分析

Table 3. Two-way analysis of variance for effects of pH and PHE on ahr, arnt and cyp1a1 gene expression of E. sinensis

基因 Gene	处理组别 Group	自由度 DF	均方差 MS	F	P
ahr	PHE	1	7.889	204.860	<0.001
	pH	2	7.760	208.258	<0.001
	PHE×pH	2	5.111	134.930	<0.001
arnt	PHE	1	8.133	314.861	<0.001
	pH	2	8.276	320.375	<0.001
	PHE×pH	2	5.697	220.565	<0.001
cyp1a1	PHE	1	51.122	1 643.494	<0.001
	pH	2	28.965	931.189	<0.001
	PHE×pH	2	4.721	151.787	<0.001

下载: 导出CSV

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