Department of Aquatic Life Medicine, Pukyong National University, 599-1, Daeyeondong, Namgu, Busan 608-737, Republic of Korea.
Fish Shellfish Immunol. 2013 Aug;35(2):476-9. doi: 10.1016/j.fsi.2013.05.001. Epub 2013 May 13.
In mammals, hypoxia-inducible factor-1 α (HIF-1α) is known to play important roles not only in oxygen homeostasis but also in innate immune responses. In this study, to assess the functional role of HIF-α in respiratory burst activity of Crassostrea gigas hemocytes, oysters were injected with HIF-α- or green fluorescent protein (GFP)-targeted-long double-stranded RNAs (dsRNAs), and at 1, 3, and 7 days post-injection, knock-down of C. gigas HIF-α expression and production of reactive oxygen species (ROS) were analyzed. Expression of HIF-α in mantle, gill, and hemocytes of C. gigas was clearly down-regulated by injection of the HIF-α-targeted-long dsRNA, but was not inhibited by the GFP-targeted-long dsRNA, indicating that HIF-α expression was suppressed through sequence-specific and systemic RNA interference (RNAi). Respiratory burst activity of hemocytes was significantly increased by administration of GFP-targeted-long dsRNA. However, knock-down of HIF-α expression led to significant decrease of chemiluminescence (CL) response of C. gigas hemocytes at 3 and 7 days post-administration of HIF-α-targeted-long dsRNA, indicating the critical role of HIF-α in activation of respiratory burst activity of oyster hemocytes.
在哺乳动物中，缺氧诱导因子-1α（HIF-1α）不仅在氧平衡中发挥重要作用，而且在先天免疫反应中也发挥重要作用。在这项研究中，为了评估 HIF-α 在牡蛎血细胞呼吸爆发活性中的功能作用，用 HIF-α 或绿色荧光蛋白（GFP）靶向长双链 RNA（dsRNA）注射牡蛎，在注射后 1、3 和 7 天，分析了 C. gigas HIF-α表达和活性氧（ROS）的产生。HIF-α 在牡蛎的套膜、鳃和血细胞中的表达明显被 HIF-α 靶向长 dsRNA 注射下调，但不受 GFP 靶向长 dsRNA 的抑制，表明 HIF-α 的表达是通过序列特异性和全身 RNA 干扰（RNAi）被抑制的。用 GFP 靶向长 dsRNA 处理可显著增加血细胞的呼吸爆发活性。然而，HIF-α 表达的敲低导致 HIF-α 靶向长 dsRNA 给药后 3 和 7 天牡蛎血细胞的化学发光（CL）反应显著降低，表明 HIF-α 在激活牡蛎血细胞呼吸爆发活性中起关键作用。
