本论文对紫外线(58μW/cm2,30 min)诱导的海参体壁自溶进行了基因表达的研究。采用TRIzol法提取海参体壁中的总RNA,以细胞色素B(Cyt B)为内标,采用反转录聚合酶链式反应(RT-PCR)对组织蛋白酶C(CC)、纤维蛋白原A(FGL)、衰老相关蛋白(SAP)和主要卵黄蛋白2(MYP2)、组织蛋白酶L(CL)、钙网蛋白(Calreticulin)、基质金属蛋白酶14(MMP14)、乙酰胆碱酯酶(Ach E)等29种分子进行基因扩增。结果显示,紫外线照射30 min后,海参体壁组织蛋白酶C、纤维蛋白原A、衰老相关蛋白和主要卵黄蛋白2的基因表达水平显著上调(p〈0.05),上调水平分别较对照组提高了71.4±44.8%、27.1±18.4%、43.7±21.6%和165.5±122.7%,而组织蛋白酶L、钙网蛋白、基质金属蛋白酶14、乙酰胆碱酯酶等基因表达没有显著变化。这些结果表明组织蛋白酶C、纤维蛋白原A、衰老相关蛋白和主要卵黄蛋白2可能参与海参体壁的自溶过程。
In this study, gene expressions for UV-induced (58 μW/cm2, 30 min) body wall autolysis of sea cucumber (Stichopusjaponicus) were investigated. Total RNA was extracted using the TRIzol method from sea cucumber body wall. Twenty-nine genes, including cathepsin C (CC), fibrinogen-like protein A (FGL), senescence-associated protein (SAP), major yolk protein 2 (MYP2), cathepsin L (CL), calreticulin, matrix metalloproteinasesl4 (MMP14), and acetylcholinesterase (ACHE), were amplified using reverse transcription-polymerase chain reaction with cytochrome B (CytB) as an internal control. The results indicated that the gene expressions of CC, FGL, SAP, and MYP2 were significantly enhanced by 71.4 ± 44.8%, 27.1 ±18.4%, 43.7 ± 21.6%, and 165.5 ± 122.7%, respectively, after UV irradiation for 30 min (p 〈 0.05). However, the gene expressions ofCL, calreticulin, MMPI4, and ACHE did not change significantly. These results suggest that CC, FGL, SAP, and MYP 2 may be involved in the autolysis of sea cucumber body wall.