嗜水气单胞菌刺激对中华绒螯蟹免疫的影响

Effects of Aeromonas hydrophila Stimulation on Immunity of Chinese Mitten Crab Eriocheir sinensis

以中华绒螯蟹为试验对象,采用常规石蜡切片和苏木精—伊红染色及实时荧光定量PCR技术分析嗜水气单胞菌刺激对其免疫组织结构及基因表达的影响。试验结果显示,与对照组相比,随着细菌刺激时间的延长,中华绒螯蟹鳃丝厚度和血细胞数量均表现为先降低后增厚增多趋势,肝小管管腔比面积、结构和细胞数量发生显著变化;脂多糖和β-1,3-葡聚糖结合蛋白表达量6 h时显著降低(P<0.05),而在12 h时极显著增加(P<0.01);Toll通路重要组件Tube表达量在3、6 h时均显著降低(P<0.05),12 h显著增加(P<0.05);Toll通路重要组件Dorsal基因表达量在3 h时显著增加(P<0.05);整合素基因表达量在感染3、6 h均显著增加(P<0.05),12 h时表现为极显著增加(P<0.01);酚氧化酶原激活酶、细胞黏附蛋白、谷胱甘肽硫转移酶基因表达量在感染12 h内均呈逐渐下降趋势;谷胱甘肽过氧化物酶基因表达量在感染6 h达到最低值,而后上调;超氧化物歧化酶基因表达量在感染6 h显著降低(P<0.05)。试验结果表明,嗜水气单胞菌刺激显著影响中华绒螯蟹鳃、肝胰腺组织结构和免疫相关基因的表达。

In this study,the influence of Aeromonas hydrophila stimulation on immune tissue structure and gene expression of Chinese mitten crab Eriocheir sinensis was analyzed by conventional paraffin section,hematoxylin-eosin staining(HE staining)and real-time fluorescence quantitative PCR.The results showed that,with the bacterial stimulation,the gill filaments thickness and the number of blood cells of Chinese mitten crab were decreased and then increased,the lumen area,structure and cell number of hepatic small tubules were changed significantly compared with the control group.The expression levels of lipopolysaccharide and beta-1,3-glucan binding protein(LGBP)were decreased significantly at 6 h(P<0.05),but increased significantly at 12 h(P<0.05).Tube gene expression level was decreased significantly at 3 h and 6 h(P<0.05),and increased significantly at 12 h(P<0.05).Dorsal expression levels were increased significantly at 3 h(P<0.05),The expression level of integrin was significantly increased at 3 h and 6 h after infection(P<0.05),and was significantly increased at 12 h(P<0.01).The expressions of prophenoloxidase activating factor(PPAF),peroxinectin(PXN)and glutathione S-transferase(GST)were found to be decreased gradually within 12 h after infection.The expression levels of glutathione peroxidase(GPx)reached the lowest value at 6 h after infection,and then was upregulated.The expression level of superoxide dismutase(SOD)was significantly decreased at 6 h after infection(P<0.05).In conclusion,Aeromonas hydrophila stimulation significantly changed the structures of the gill and hepatopancreas and the expression levels of immune-related genes in Chinese mitten crab.