摘要
微塑料(microplastics,MPs)是常见的污染物,而硫酸铜(CuSO4)是水产养殖中常用的消毒剂,水体中的铜离子(Cu^(2+))可被MPs吸附。为探究MPs和Cu^(2+)单一及联合暴露对中华绒螯蟹(Eriocheir sinensis)肠道组织的影响机制,本研究选择0.4 mg/L MPs和0.1 mg/L Cu^(2+)作为实验浓度,设置4个处理组:0.4 mg/L MPs暴露组(M组)、0.1 mg/L Cu^(2+)暴露组(C组)、0.4 mg/L MPs+0.1 mg/L Cu^(2+)联合暴露组(MC组)和空白对照组(D组),探究MPs和Cu^(2+)单一及联合暴露21 d后对中华绒螯蟹肠道的影响。采用RNA-Seq技术对肠道开展转录组测序,筛选差异表达基因(DEGs)进行KEGG富集等生物信息学分析。结果显示,与对照组相比,M组包括1650个上调DEGs和1874个下调DEGs;C组包括3797个上调DEGs和1073个下调DEGs;MC组包括1492个上调DEGs和1305个下调DEGs。与抗氧化、免疫和能量代谢相关的DEGs,包括过氧化氢酶(cat)、三结构域蛋白(trim)、Toll样受体(tlr)、细胞色素P450(cyp450)、碳酸酐酶(ca)等在M vs D、C vs D、MC vs D中变化显著。KEGG富集分析显示,DEGs主要富集于氧化磷酸化、谷胱甘肽代谢、细胞色素P450对异生素的代谢等通路。结果表明,MPs与Cu^(2+)单一及联合暴露导致中华绒螯蟹的抗氧化、免疫和能量代谢系统失衡,MPs和Cu^(2+)具有协同毒性作用,研究结果为阐明MPs和Cu^(2+)暴露对中华绒螯蟹的影响机制提供了参考。
Over 80% of marine litter is composed of plastic waste,which reaches the oceans via atmospheric transport,surface runoff,and human activities such as shipping and fishing.These plastics degrade into microplastics(MPs)—prevalent environmental pollutants<5 mm,which possess a“carrier effect”,enabling them to adsorb contaminants such as heavy metals.CuSO_(4)—frequently used in aquaculture to manage diseases and cyanobacterial blooms—can introduce excess Cu^(2+)into aquatic environments,adversely affecting water quality and aquatic life.The Chinese mitten crab(Eriocheir sinensis)is vital to freshwater aquaculture in China.Mature E.sinensis migrate to the Yangtze River estuary annually for reproduction;however,estuarine ecosystems are becoming increasingly disturbed,making mitten crabs vulnerable to MPs and heavy metal contamination.The intestinal tract,which directly interacts with ingested pollutants,is particularly susceptible.Previous studies have examined MP or Cu^(2+)effects on E.sinensis independently,however,their combined effects remain underexplored.Here,MPs(0.4 mg/L)and Cu^(2+)(0.1 mg/L)were selected as the experimental concentrations with four treatment groups:Group M(0.4 mg/L MP-exposed);Group C(0.1 mg/L Cu^(2+)-exposed);Group MC(0.4 mg/L MP+0.1 mg/L Cu^(2+)combined-exposed;and Group D(blank control group)to investigate the effects of MPs and Cu^(2+)on the intestinal tract of E.sinensis after 21 days of single and combined exposure.Transcriptome sequencing of the intestinal tissues was conducted using Illumina's high-throughput platform,generating 197,908,972 raw reads.Post-quality filtering yielded clean reads across the four groups,with 94.71%-95.48%Q30 scores.In total,109,644 transcripts were identified,with 68,005 exceeding 1800 bp in length.Differential expression analysis revealed 1,650 and 1,874,3,797 and 1,073,and 1,492 and 1,305 upregulated and downregulated differentially expressed genes(DEGs)in Groups M,C,MC,respectively.Notably,DEGs associated with antioxidant defense,immune response,and energy metabolism differed significantly among the comparison groups.Catalase(cat)and peroxiredoxin(prdx)were downregulated,whereas trim,toll-like receptor(tlr)and complement component 1(c1)were upregulated in the immune system.Cytochrome P450(cyp450)was significantly downregulated across the treatment groups.Energy metabolism DEGs also varied,with carbonic anhydrase(ca)up-regulated and vesicular adenosine triphosphatase(v-atpase)down-regulated.KEGG pathway enrichment analysis indicated that in Groups M vs D,1,594 DEGs in the intestine were mapped to 333 pathways.Among these,34 pathways were significantly enriched,including oxidative phosphorylation and glutathione metabolism.In Groups C vs D,2,445 DEGs in the intestine were mapped to 340 pathways.Among these,nine pathways were significantly enriched,primarily involving DNA replication and ABC transporters.In Groups MC vs D,1,198 DEGs in the intestine were mapped to 326 pathways.Among these,14 pathways were significantly enriched,including the complement and coagulation cascades and metabolism of xenobiotics by cytochrome P450.The DEGs were predominantly enriched in pathways related to oxidative phosphorylation,glutathione metabolism,xenobiotic metabolism by cytochrome P450,and ABC transporters.These findings indicate that both individual and combined exposure to MPs and Cu^(2+)disrupts the antioxidant,immune,and energy metabolic systems of E.sinensis.The glutathione metabolic pathway was particularly inhibited in Groups M,C,and MC.MPs and Cu^(2+)may affect the expression of cyp450 and related genes(ugt),potentially compromising the immune function of E.sinensis.Notably,ugt was significantly upregulated in Groups M vs D and downregulated in Groups MC vs D.The number of DEGs linked to the oxidative phosphorylation pathway varied across comparisons,with 41 DEGs in Groups M vs D and seven in Groups C vs D and MC vs D being enriched in this pathway.MPs may affect the oxidative phosphorylation pathway by inhibiting the expression of cox and atpase genes.In contrast,exposure to Cu^(2+)alone and co-exposure to MPs had comparatively smaller impacts on this pathway.The mechanisms underlying the stress response of E.sinensis to MPs and Cu^(2+)exposure were further elucidated in this study.The results of this study revealed the mechanisms of MPs and Cu^(2+)'s effects on the antioxidant,immune defence,and energy metabolism of Eriocheir sinensis,which provides an important theoretical basis for the study of E.sinensis culture and environmental toxicity.
作者
徐建
冯广朋
严云志
陈锦辉
吴建辉
杨祖长
XU Jian;FENG Guangpeng;YAN Yunzhi;CHEN Jinhui;WU Jianhui;YANG Zuchang(School of Ecology and Environment,Anhui Normal University,Wuhu 241002,China;Shanghai Yangtze Estuary Fishery Resources Proliferation and Ecological Restoration Engineering Technology Research Center,East China Sea Fisheries Research Institute,Chinese Academy of Fishery Sciences,Shanghai 200090,China;Joint Laboratory for Monitoring and Conservation of Aquatic Living Resources in the Yangtze Estuary,Shanghai Aquatic Wildlife Conservation and Research Center,Shanghai 200092,China)
出处
《渔业科学进展》
北大核心
2025年第2期173-188,共16页
Progress in Fishery Sciences
基金
长江口南支东部水域中华绒螯蟹等重要物种变化趋势预测实施项目(11N42500663520242801)资助。
关键词
中华绒螯蟹
微塑料
铜
转录组
能量代谢
免疫
Eriocheir sinensis
Microplastics
Copper
Transcriptome
Energy metabolism
Immunity
