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一种潜在的抗凋亡调节:海参热应激和夏眠期间热休克蛋白70与凋亡诱导因子线粒体1的相互作用。

A potential antiapoptotic regulation: The interaction of heat shock protein 70 and apoptosis-inducing factor mitochondrial 1 during heat stress and aestivation in sea cucumber.

作者信息

Wang Shasha, Li Xingke, Chen Muyan, Storey Kenneth B, Wang Tianming

机构信息

Key Laboratory of Mariculture, Ministry of Education, Ocean University of China, Qingdao, PR China.

Institute of Biochemistry, Carleton University, Ottawa, ON, Canada.

出版信息

J Exp Zool A Ecol Integr Physiol. 2018 May 28. doi: 10.1002/jez.2180.

Abstract

The sea cucumber (Apostichopus japonicus) has become a good model organism for studying environmentally induced aestivation in marine invertebrates. A characteristic feature of aestivation in this species is the degeneration of the intestine. In the current study, we hypothesized that energy conservation and cytoprotective strategies need to be coordinated in the intestine to ensure long-term survival during aestivation, and there was potential relationship between heat shock protein 70 (HSP70) and apoptosis-inducing factor mitochondrial 1 (AIFM1) during extreme environmental stress. AIFM1 is a bifunctional flavoprotein that is involved in the caspase-independent activation of apoptosis. The gene and protein expression profiles of AjAIFM1 and AjHSP70 in intestinal tissue during aestivation were analyzed and results showed an inverse correlation between them, AjAIFM1 being suppressed during aestivation whereas AjHSP70 was strongly upregulated. Comparable responses were also seen when intestinal cells were isolated and analyzed in vitro for responses to heat stress at 25°C (a water temperature typical during aestivation), compared with 15°C control cells. Combined with co-immunoprecipitation studies in vivo and in vitro, our results suggested that AjHSP70 protein may have potential interaction with AjAIFM1. To determine the influence of heat stress on apoptotic rate of intestinal cells, we also assessed the DNA fragmentation by terminal deoxynucleotide transferase-mediated dUTP nick-end labeling assay, and results also supported a potential antiapoptotic response in sea cucumber during heat stress. This type of cytoprotective mechanism could be used to preserve the existing cellular components during long-term aestivation in sea cucumber.

摘要

海参(刺参)已成为研究海洋无脊椎动物环境诱导夏眠的良好模式生物。该物种夏眠的一个特征是肠道退化。在本研究中,我们假设在夏眠期间,肠道需要协调能量守恒和细胞保护策略以确保长期存活,并且在极端环境压力下热休克蛋白70(HSP70)与凋亡诱导因子线粒体1(AIFM1)之间存在潜在关系。AIFM1是一种双功能黄素蛋白,参与不依赖半胱天冬酶的凋亡激活。分析了夏眠期间肠道组织中刺参AIFM1(AjAIFM1)和刺参HSP70(AjHSP70)的基因和蛋白质表达谱,结果显示它们之间呈负相关,夏眠期间AjAIFM1受到抑制,而AjHSP70强烈上调。当分离肠道细胞并在体外分析其对25°C(夏眠期间典型水温)热应激的反应时,与15°C对照细胞相比,也观察到了类似的反应。结合体内和体外的免疫共沉淀研究,我们的结果表明AjHSP70蛋白可能与AjAIFM1存在潜在相互作用。为了确定热应激对肠道细胞凋亡率的影响,我们还通过末端脱氧核苷酸转移酶介导的dUTP缺口末端标记法评估了DNA片段化,结果也支持海参在热应激期间存在潜在的抗凋亡反应。这种细胞保护机制可用于在海参长期夏眠期间保存现有的细胞成分。

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