Marine Applied Microbes and Aquatic Organism Disease Control Lab, Department of Aquatic Biomedical Sciences, School of Marine Biomedical Sciences, College of Ocean Sciences & Marine and Environmental Research Institute, Jeju National University, Jeju 690-756, South Korea.
Fish Shellfish Immunol. 2010 Apr;28(4):511-6. doi: 10.1016/j.fsi.2009.12.005. Epub 2009 Dec 16.
The innate immunity and expression profiles of cathepsins D were determined in the goldfish (Carassius auratus) tissues after challenge with a fish pathogen Aeromonas hydrophila. The innate immunity of reactive oxygen species (ROS) and reactive nitrogen species (RNS) were determined by peripheral blood leucocytes. Blood and tissue samples of the muscle, gills, liver, kidney, heart, spleen, and intestine were sampled at 1, 3, 6 and 12 h post-infection for cathepsin D expression by semi-quantitative RT-PCR. The ROS and RNS production did not significantly increase at 1 h post-challenged goldfish. However, the ROS and RNS production was significantly increased after 3 h post-challenged fish compared to the control. The cathepsin D expression was found very low in muscle and kidney of the control fish, other tissues was not found the expression. A similar pattern was found in goldfish at 1 h post-challenge with A. hydrophila. However, at 3 h post-challenge goldfish, the cathepsin D expression was high only in the heart. At 6 h post-challenge goldfish, the cathepsin D expression was seen high all the tissues, except in the spleen. However, the expression was decreased at 12 h post-infection samples. This result was suggested that the goldfish infected with A. hydrophila decreased the innate immunity level in peripheral blood and expressed the cathepsin D in tissues.
本研究检测了嗜水气单胞菌感染金鱼后组织中组织蛋白酶 D 的先天免疫和表达谱。通过外周血白细胞测定活性氧（ROS）和活性氮（RNS）的先天免疫。在感染后 1、3、6 和 12 h 时，采集金鱼的肌肉、鳃、肝、肾、心、脾和肠的血液和组织样本，通过半定量 RT-PCR 检测组织蛋白酶 D 的表达。在 challenged 后的 1 h，ROS 和 RNS 的产生没有显著增加。然而，与对照组相比，在 challenged 后的 3 h，ROS 和 RNS 的产生显著增加。在对照组的金鱼肌肉和肾脏中发现组织蛋白酶 D 的表达非常低，其他组织未发现表达。在 challenged 后的 1 h 也发现了类似的模式。然而，在 challenged 后的 3 h，组织蛋白酶 D 的表达仅在心脏中较高。在 challenged 后的 6 h，组织蛋白酶 D 的表达在除脾脏以外的所有组织中都较高。然而，在感染后 12 h 的样本中，表达水平下降。这一结果表明，感染嗜水气单胞菌的金鱼外周血的先天免疫水平降低，并在组织中表达组织蛋白酶 D。
