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 下短暂预处理冷休克 3 分钟被证明是热休克蛋白 70 (HSP70) 的诱导剂，并作为分子伴侣改善了应激耐受性。随着空气暴露耐受性的提高，冷休克后虾血细胞中HSP70的mRNA和蛋白水平上调。成功建立了 HSP70 RNA 干扰 (RNAi) 基因敲低和重组 HSP70 (rHSP70) 注射，以研究 HSP70 在应对空气暴露压力中的作用。与冷休克处理的虾（对照，90%）相比，接受 rHSP70 的虾在空气暴露下显示出提高的存活率（80%），但没有显着差异（p  > 0.05），但 HSP70 敲低的虾的存活率显着降低（ 62%, p < 0.05)。富集 HSP70 的虾（即冷休克和 rHSP70 注射）的活性氧（ROS）含量、细胞色素c的相对表达、caspase-3 活性和细胞凋亡率显着低于 HSP70 基因敲除虾（ p  < 0.05） . 结果表明，HSP70可通过冷休克诱导，并通过清除细胞内ROS、抑制细胞色素c表达、抑制线粒体释放和灭活caspase-3来阻断细胞凋亡途径，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冷激机制的认识。