Ahn Meejung, Ko Hyun Ju, Kim Jeongtae, Jang Yeounghwan, Shin Taekyun
College of Veterinary Medicine and Veterinary Medical Research Institute, Jeju National University, Jeju 63243, Republic of Korea.
It's Hanbul Co., Ltd. Research Institute 4, Nonhyeon-dong 249, Gangnam-gu, Seoul 06101, Republic of Korea.
Parasite. 2018;25:12. doi: 10.1051/parasite/2018013. Epub 2018 Mar 13.
Kudoa septempunctata (Myxosporea, Multivalvulida) is a parasite of the trunk muscle of cultured olive flounder (Paralichthys olivaceus). We investigated whether K. septempunctata genotype ST3 spores induce cell damage and the secretion of inflammatory mediators in Caco-2 cells, which exhibit characteristics similar to human intestinal epithelial cells. Purified K. septempunctata spores were heated at 95 °C for 5 min. Lactate dehydrogenase (LDH) release was measured to determine the efficacy of denaturation. Naïve and heated spores, lipopolysaccharide (positive control) and vehicle (negative control) were added to Caco-2 cells. Cells were subjected to the cytotoxic LDH assay and western blot analysis to examine the expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX)-2. Supernatants were collected to measure nitric oxide (NO) and prostaglandin E2 (PGE). Most spores were denaturated by heating, and the spore morphology was found to be wrinkled with shell valves and polar capsules. In addition, cytotoxicity and inflammatory mediators, such as NO, PGE, iNOS, and COX-2, remained unchanged in Caco-2 cells following exposure to naïve and heated spores compared with the positive controls. Collectively, the findings of this study imply that spores of K. septempunctata genotype ST3 do not cause inflammation in Caco-2 cells.
七斑库道虫（黏孢子虫纲，多壳目）是养殖牙鲆（ Paralichthys olivaceus）躯干肌肉的一种寄生虫。我们研究了七斑库道虫基因型ST3孢子是否会诱导与人类肠道上皮细胞具有相似特征的Caco-2细胞发生细胞损伤及炎性介质的分泌。将纯化的七斑库道虫孢子在95°C加热5分钟。通过测量乳酸脱氢酶（LDH）释放来确定变性效果。将未处理的和加热后的孢子、脂多糖（阳性对照）和赋形剂（阴性对照）添加到Caco-2细胞中。对细胞进行细胞毒性LDH测定和蛋白质印迹分析，以检测诱导型一氧化氮合酶（iNOS）和环氧化酶（COX）-2的表达。收集上清液以测量一氧化氮（NO）和前列腺素E2（PGE）。大多数孢子通过加热变性，发现孢子形态带有壳瓣和极囊且有褶皱。此外，与阳性对照相比，Caco-2细胞在暴露于未处理的和加热后的孢子后，细胞毒性和炎性介质如NO、PGE、iNOS和COX-2保持不变。总体而言，本研究结果表明七斑库道虫基因型ST3的孢子不会在Caco-2细胞中引发炎症。
