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致病性粪肠球菌对脊尾白虾(Exopalaemon carinicauda)免疫相关基因表达的影响
段亚飞,李 健,李吉涛,张 喆,刘 萍
1.农业部南海渔业资源开发利用重点实验室 中国水产科学研究院南海水产研究所 广州 510300;2.农业部海洋渔业可持续发展重点实验室 中国水产科学研究院黄海水产研究所 青岛 266071;3.青岛海洋科学与技术国家实验室 海洋渔业科学与食物产出过程功能实验室 青岛 266071
摘要:
为了研究致病性粪肠球菌(Enterococcus faecalis)对脊尾白虾(Exopalaemon carinicauda)免疫相关基因表达的影响,分别对健康脊尾白虾注射生理盐水和粪肠球菌,测定了不同时间点血细胞和肝胰腺中酚氧化酶原(proPO)、C型凝集素(CTL)、超氧化物歧化酶(SOD)、组织蛋白酶D (Cat D)和组织蛋白酶L (Cat L)基因的相对表达量变化。结果显示,与对照组相比,感染粪肠球菌后,血细胞和肝胰腺中proPO基因表达量均于6 h达到最大值(P<0.05),随后逐渐降低至对照组水平;血细胞和肝胰腺中CTL基因表达量分别于12 h和6 h上升至最大值,随后逐渐降低;血细胞中CTL基因表达量于72 h虽有降低,但仍显著高于对照组(P<0.05);血细胞和肝胰腺中SOD基因表达量分别于6 h和3 h达到最大值,随后逐渐降低,并于72 h达到正常水平;血细胞和肝胰腺中Cat D基因表达量变化趋势基本一致,呈现先降低后升高、然后显著降低再升高的趋势;血细胞和肝胰腺中Cat L基因表达量分别于6 h和3 h显著降低,随后逐渐升高至最大值,并于72 h达到对照组水平。研究表明,长时间的粪肠球菌感染对脊尾白虾免疫相关基因表达影响显著,本研究所用免疫相关基因均可作为粪肠球菌感染疾病的监测指标。
关键词:  脊尾白虾  粪肠球菌  酚氧化酶原  C型凝集素  超氧化物歧化酶  组织蛋白酶
DOI:10.11758/yykxjz.20160507001
分类号:
基金项目:
The Expression Profiles of Immunity-Related Genes After Pathogenic Enterococcus faecalis Challenge in Exopalaemon carinicauda
DUAN Yafei1, LI Jian2,3, LI Jitao4,5, ZHANG Zhe1, LIU Ping4,5
1.Key Laboratory of South China Sea Fishery Resources Exploitation & Utilization, Ministry of Agriculture, South China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou 510300;2.Key Laboratory of Sustainable Development of Marine Fisheries, Ministry of Agriculture, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071;3.Laboratory for Marine Fisheries Science and Food Production Processes, Qingdao National Laboratory for Marine Science and Technology, Qingdao 266071;4.Key Laboratory of Sustainable Development of Marine Fisheries, Ministry of Agriculture, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266072;5.Laboratory for Marine Fisheries Science and Food Production Processes, QiǙ地碃Ǚ෠溺Ǚ塰碃Ǚ᥀溺Ǚ妰碃Ǚ嚀溽Ǚ�碃Ǚ蓰溽Ǚ挈碃Ǚ诰溽Ǚ�碃Ǚꀐ溽Ǚ憠碃Ǚꊰ溽Ǚ�碃Ǚ꣐溽Ǚ�碃Ǚะ滃Ǚ�碃Ǚ
Abstract:
In this study, we investigated the effect of pathogenic Enterococcus faecalis challenge on the non-specific immunity-related gene expression of Exopalaemon carinicauda. The expression levels of prophenoloxidase (proPO), C-type lectin (CTL), superoxide dismutase (SOD), cathepsin D (Cat D) and cathepsin L (Cat L) in hemocytes and hepatopancreas were measured with quantitative real-time PCR at different sampling time points after healthy E. carinicauda were injected with 0.86% sterile saline (control) or pathogenic E. faecalis. Compared to the control group, after infected with E. faecalis, the relative expression levels of proPO gene in hemocytes and hepatopancreas both increased to the highest at 6 h (P<0.05), and then decreased gradually back to normal. The relative expression levels of CTL gene in hemocytes and hepatopancreas increased to the peak value at 12 h and 6 h respectively (P<0.05), and then decreased gradually. Although the relative expression levels of CTL gene in hemocytes decreased at 72 h, it was still significantly higher than the control (P<0.05). The relative expression levels of SOD gene in hemocytes and hepatopancreas both increased to the highest at 6 h and 3 h respectively (P<0.05), then decreased gradually back to normal at 72 h. The dynamic expression of Cat D gene in hemocytes and hepatopancreas was consistent, which was a significant decreased in the beginning followed by an increase. After the decrease of expression at 12 h, it was increased significantly again. The Cat L gene expression in hemocytes and hepatopancreas both decreased significantly at 6 h and 3 h respectively (P<0.05), then it increased to the highest and recovered to the control level at 72 h. The results indicated that pathogenic E. faecalis challenge affected the immune status of E. carinicauda probably by changing the immunity-related gene expression, and that proPO, CTL, SOD, Cat D and Cat L were all sensitive to E. faecalis infection.
Key words:  Exopalaemon carinicauda  Enterococcus faecalis  Prophenoloxidase  C-type lectin  Superoxide dismutase  Cathepsin