Ishiwata Kenji, Watanabe Naohiro
Department of Tropical Medicine, The Jikei University School of Medicine, 3-25-8, Nishi-shinbashi, Minato-ku, Tokyo 105-8461, Japan.
Exp Parasitol. 2007 Sep;117(1):80-6. doi: 10.1016/j.exppara.2007.03.019. Epub 2007 Apr 4.
Gastrointestinal nematodes require energy for active establishment in the gut against intestinal flow and peristaltic motion. In this study we employed CellTiter-Glo Luminescent Cell Viability Assay to measure the ATP value of individual adult Nippostrongylus brasiliensis during the course of immune-mediated expulsion from the small intestine in rats. The ATP values of adult worms taken from the lumen of the distal small intestine were lower than worms collected from the lumen of the proximal small intestine. Moreover, values from worms in the lumen of the proximal small intestine were lower than those from worms in the mucosa, the preferred site of adult N. brasiliensis. The reduction of ATP values in worms from each region was observed not only at expulsion phase, but also at established phases of the infection suggesting that energy metabolism of the parasites is independent of host immune response. When adult worms with low ATP values on day 12 post-infection were implanted surgically into the small intestine of naïve rats, the worms re-established in recipients and completely restored the ATP values. Short in vitro culture of adult worms under low oxygen tension resulted in low ATP value in the worms. These results suggested that adult worms were dislodged from their preferred site by intact energy metabolism activity.
胃肠道线虫需要能量以在肠道中逆着肠内容物流动和蠕动运动而主动定殖。在本研究中,我们采用CellTiter-Glo发光细胞活力测定法来测量成年巴西日圆线虫在大鼠小肠免疫介导排虫过程中的ATP值。取自远端小肠肠腔的成虫的ATP值低于取自近端小肠肠腔的成虫。此外,近端小肠肠腔中虫体的ATP值低于黏膜中虫体的ATP值,而黏膜是巴西日圆线虫成虫的偏好定殖部位。不仅在排虫阶段,而且在感染的定殖阶段,都观察到来自每个区域的虫体的ATP值降低,这表明寄生虫的能量代谢独立于宿主免疫反应。当将感染后第12天ATP值较低的成虫手术植入未感染大鼠的小肠时,这些虫体在受体中重新定殖并完全恢复了ATP值。在低氧张力下对成虫进行短时间体外培养导致虫体ATP值降低。这些结果表明,完整的能量代谢活动使成虫从其偏好的定殖部位被驱离。