Chen Honglang, Song Lijun, Li Guixian, Chen Wenfeng, Zhao Shumin, Zhou Ruoxia, Shi Xiaoying, Peng Zhenying, Zhao Wenchang
School of Pharmacy, Guangdong Medical University, No. 1, Xincheng Road of Songshan Lake Science and Technology Industry Park, Dongguan, 523808, Guangdong Province, China.
Institution of Traditional Chinese Medicine and New Pharmacy Development, Guangdong Medical University, Dongguan, 523808, Guangdong Province, China.
Virus Genes. 2017 Jun;53(3):367-376. doi: 10.1007/s11262-017-1444-0. Epub 2017 Mar 13.
Rotavirus (RV) is the most common cause of severe gastroenteritis and fatal dehydration in human infants and neonates of different species. However, the pathogenesis of rotavirus-induced diarrhea is poorly understood. Secretory diarrhea caused by rotavirus may lead to a combination of excessive secretion of fluid and electrolytes into the intestinal lumen. Fluid absorption in the small intestine is driven by Na-coupled transport mechanisms at the luminal membrane, including Na/H exchanger (NHE). Here, we performed qRT-PCR to detect the transcription of NHEs. Western blotting was employed for protein detection. Furthermore, immunocytochemistry was used to validate the NHE's protein expression. Finally, intracellular Ca concentration was detected by confocal laser scanning microscopy. The results demonstrated that the NHE6 mRNA and protein expressed in the human colon adenocarcinoma cell line (Caco-2). Furthermore, RV-Wa induced decreased expression of the NHE1 and NHE6 in Caco-2 cell in a time-dependent manner. In addition, intracellular Ca concentration in RV-Wa-infected Caco-2 cells was higher than that in the mock-infected cells. Furthermore, RV-Wa also can downregulate the expression of calmodulin (CaM) and calmodulin kinase II (CaMKII) in Caco-2 cells. These findings provides important insights into the mechanisms of rotavirus-induced diarrhea. Further studies on the underlying pathophysiological mechanisms that downregulate NHEs in RV-induced diarrhea are required.
轮状病毒（RV）是人类婴儿和不同物种新生儿中严重肠胃炎和致命脱水的最常见病因。然而，轮状病毒诱导腹泻的发病机制尚不清楚。轮状病毒引起的分泌性腹泻可能导致液体和电解质过度分泌到肠腔中。小肠中的液体吸收是由肠腔膜上的钠偶联转运机制驱动的，包括钠/氢交换体（NHE）。在此，我们进行了定量逆转录聚合酶链反应（qRT-PCR）以检测NHEs的转录。采用蛋白质印迹法进行蛋白质检测。此外，免疫细胞化学用于验证NHE的蛋白质表达。最后，通过共聚焦激光扫描显微镜检测细胞内钙浓度。结果表明，NHE6 mRNA和蛋白质在人结肠腺癌细胞系（Caco-2）中表达。此外，RV-Wa以时间依赖性方式诱导Caco-2细胞中NHE1和NHE6的表达降低。此外，RV-Wa感染的Caco-2细胞中的细胞内钙浓度高于模拟感染细胞。此外，RV-Wa还可下调Caco-2细胞中钙调蛋白（CaM）和钙调蛋白激酶II（CaMKII）的表达。这些发现为轮状病毒诱导腹泻的机制提供了重要见解。需要进一步研究RV诱导腹泻中下调NHEs的潜在病理生理机制。
