College of Biosystems Engineering and Food Science, Zhejiang Key Laboratory of Agro-Food Processing, Key Laboratory of Agro-Products Postharvest Handling of Ministry of Agriculture and Rural Affairs, Zhejiang University, Hangzhou, 310058, China.
Department of Chemistry, Zhejiang University, Hangzhou, 310027, China.
Dev Comp Immunol. 2021 Jan;114:103844. doi: 10.1016/j.dci.2020.103844. Epub 2020 Aug 28.
Brief pretreatment of cold shock at 13 °C for 3 min proved to be an inducer of heat shock protein 70 (HSP70) and improved stress tolerance as a molecular chaperone. With the improvement of air exposure tolerance, HSP70 in shrimp hemocytes was upregulated in mRNA and protein levels after cold shock. Both HSP70 RNA interference (RNAi) gene knockdown and recombinant HSP70 (rHSP70) injection were successfully established in order to investigate the role of HSP70 in response to air exposure stress. Shrimp receiving rHSP70 showed an improved survival rate (80%) with no significant difference (p > 0.05) compared to cold shock treated shrimp (control, 90%) under air exposure, but the survival rate of HSP70-knockdown shrimp was significantly lower (62%, p < 0.05). Reactive oxygen species (ROS) content, relative expression of cytochrome c, caspase-3 activity, and apoptosis rate in hemocytes of HSP70 enriched shrimp (i.e., cold shock and rHSP70 injection) were significantly lower (p < 0.05) than HSP70-knockdown shrimp. Results suggested that HSP70 could be induced by cold shock and contributed to improve the tolerance of shrimp suffering air exposure by blocking the apoptosis pathway through scavenging intracellular ROS, inhibiting cytochrome c expression, inhibiting release from mitochondria, and inactivating caspase-3. This work updates the understanding of cold shock mechanism in water-free transportation of aquatic animals.
在 13°C 下进行 3 分钟的短暂预处理被证明是热休克蛋白 70（HSP70）的诱导物，并作为分子伴侣提高了应激耐受性。随着空气暴露耐受性的提高，虾血细胞中的 HSP70 在 mRNA 和蛋白质水平上均上调。为了研究 HSP70 在应对空气暴露应激中的作用，成功建立了 HSP70 的 RNA 干扰（RNAi）基因敲低和重组 HSP70（rHSP70）注射。接受 rHSP70 的虾在空气暴露下的存活率（80%）与冷休克处理的虾（对照，90%）相比没有显著差异（p>0.05），但 HSP70 敲低虾的存活率显著降低（62%，p<0.05）。富含 HSP70 的虾（即冷休克和 rHSP70 注射）的血细胞中活性氧（ROS）含量、细胞色素 c 的相对表达、caspase-3 活性和凋亡率均显著降低（p<0.05）。结果表明，HSP70 可被冷休克诱导，并通过清除细胞内 ROS、抑制细胞色素 c 表达、抑制线粒体释放和失活 caspase-3 来阻止凋亡途径，从而提高虾对空气暴露的耐受性。这项工作更新了对水生动物无水运输中冷休克机制的理解。
