Institute of Parasitology, Faculty of Veterinary Medicine, University of Leipzig, An den Tierkliniken 35, 04103, Leipzig, Germany.
Albrecht Daniel Thaer Institute, An den Tierkliniken 29, 04103, Leipzig, Germany.
Parasitol Res. 2019 Dec;118(12):3429-3441. doi: 10.1007/s00436-019-06471-y. Epub 2019 Oct 31.
The parasite Cryptosporidium parvum Tyzzer 1912 destroys parts of the intestinal brush border membrane which is important for the uptake of nutrients like glucose. In this study, glucose transport mechanisms of the host cells (IPEC-J2 cells) infected by C. parvum were investigated. The mRNA expression levels of glucose transporters (GLUT) 1 and 2 and Na-coupled glucose transporter (SGLT) 1 were compared in infected and uninfected cells over an infection time of 24-96 h by RT-qPCR. Furthermore, the protein expression of SGLT 1 and GLUT 2 was quantified in western blot studies. While the protein expression of SGLT 1 was not altered in infected cells, mRNA expression of SGLT 1 and GLUT 1 was significantly increased 24 h p. i. and decreased 96 h p. i. The mRNA expression of GLUT 2 was significantly decreased 24 h, 72 h, and 96 h p. i. and also correlated significantly with the infection dose at 72 h p. i. In contrast to that, the protein expression of GLUT 2 was significantly increased 48 h p. i., associated with a significantly higher intracellular glucose level in infected cells compared with control cells at that time point of infection. This points to an adaptation of the host cells' glucose uptake taking place in the acute phase of the infection. A better understanding of these molecular mechanisms following a C. parvum infection may probably lead to an improvement of therapy strategies in the future.
寄生虫微小隐孢子虫（Cryptosporidium parvum）1912 年破坏了对葡萄糖等营养物质吸收很重要的肠刷状缘膜的部分。在本研究中，研究了宿主细胞（IPEC-J2 细胞）感染微小隐孢子虫后葡萄糖转运机制。通过 RT-qPCR，在感染后 24-96 小时内比较了感染和未感染细胞中葡萄糖转运蛋白（GLUT）1 和 2 和 Na 偶联葡萄糖转运蛋白（SGLT）1 的 mRNA 表达水平。此外，还通过 Western blot 研究定量了 SGLT1 和 GLUT2 的蛋白表达。虽然感染细胞中 SGLT1 的蛋白表达没有改变，但 SGLT1 和 GLUT1 的 mRNA 表达在感染后 24 小时显着增加，96 小时后显着降低。GLUT2 的 mRNA 表达在感染后 24 小时、72 小时和 96 小时显着降低，与 72 小时时的感染剂量也显着相关。相比之下，GLUT2 的蛋白表达在感染后 48 小时显着增加，与感染细胞在该时间点的细胞内葡萄糖水平显着高于对照细胞相关。这表明宿主细胞的葡萄糖摄取在感染的急性期发生了适应性变化。更好地了解微小隐孢子虫感染后的这些分子机制可能会在未来改善治疗策略。
