摘要
运用RT-PCR技术探索拟穴青蟹(Scylla paramamosain)注射免疫增强剂——大黄多糖(rhubarb polysaccharide,RP)后11个免疫相关基因的表达变化情况。结果发现,大黄多糖刺激拟穴青蟹4 d后,血细胞中4个基因(丝氨酸蛋白酶抑制因子、过氧化氢酶、过氧化物还原酶和酚氧化酶原)表达水平分别上调1.1倍、4.1倍、1.3倍和2.1倍,与对照组相比,存在极显著(P<0.01)或显著性差异(P<0.05)。肝胰腺中5个基因表达变化显著(P<0.05),其中丝氨酸蛋白酶抑制因子、过氧化物还原酶和抗脂多糖因子3个基因表达分别上调2.1倍、4.8倍和1.7倍;过氧化氢酶和溶菌酶仅在实验组中表达,未见其在对照组表达。由此推测,丝氨酸蛋白酶抑制因子、过氧化氢酶、过氧化物还原酶、酚氧化酶原、抗脂多糖因子和溶菌酶6个基因可能与拟穴青蟹免疫增强有关。
The mud crab （Scylla paramamosain） is widely cultured on the southeast coast of China. Unfortunately, the crab industry has suffered significant economic losses in the past 10 years due to viral, bacterial, and parasitic diseases. Researchers have suggested that using immunostimulants would help establish a crab disease control strategy. However, little information on immune-enhancing proteins in crabs is available. In this study, a quantitative real-time reverse transcription-polymerase chain reaction （RT-PCR） analysis was carried out to investigate whether 11 reported genes, including metallothionein （Met）, serine protease inhibitor （Spi）, lysozyme （Lys）, cmstin （Cru）, catalase （Cat）, peroxide reductase （Per）, prophenoloxidase （Pro）, anti-lipopolysaccharide factor （All）, Scylla serrata anti-microbial Scy2 pre- cursor, serine proteinase （Sek）, and peroxinectin （Per） are associated with enhancing immunity in S. paramamosain treated with rhubarb polysaccharides for 4 days. The results indicate that the mean hemocyte mRNA expression values of Spi, Cat, Per, and Pro increased about 1.1-, 4.1-, 1.2-, and 2.1-fold, respectively. At the same time, the Spi, Cat, Per, Lys, and A/f genes in the hepatopancreas were significantly upregulated. Of these, Spi, Per, and A/f were upregulated 2.1-, 4.8-, and 1.7-fold, respectively. The Lys and Cat genes were only expressed in the experimental group. This result was different from our previous findings that five related immune-enhancing proteins, including hemocyanin, chy- motrypsin, cryptocyanin, C-type lectin receptor, and ferritin protein, were identified in S. paramamosain using proteo- mic and RT-PCR strategies. The discrepancy probably reflects differences between the two research approaches or an uni- dentified post-transcriptional mechanism in these related immune-enhancing proteins. Considering recent evidence that the Spi, Cat, Per, Pro, Lys, and Alf genes play significant roles in invertebrate immune defense, it seems likely that these six significantly upregulated genes in hemocytes and hepatopancreas may be related to enhancing the crab＇s immune response. These results will help reveal the immune enhancement mechanism in S. paramamosain and will be important for immunological prevention of crab diseases.
出处
《中国水产科学》
CAS
CSCD
北大核心
2015年第3期387-392,共6页
Journal of Fishery Sciences of China
基金
国家自然科学基金项目(31372558)
教育部新世纪优秀人才支持计划项目(NCET-11-0922)
广东省自然科学基金项目(S2013010015190)
广东省高等学校科技创新重点项目(2012CXZD0025)
广东省高等学校高层次人才项目(2011)
关键词
拟穴青蟹
大黄多糖
RT-PCR
免疫增强相关基因
Scylla paramamosain
rhubarb polysaccharide
RT-PCR
immune enhancement-related gene
