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Human rotavirus strain Wa downregulates NHE1 and NHE6 expressions in rotavirus-infected Caco-2 cells

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Abstract

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 Ca2+ 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 Ca2+ 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.

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Acknowledgements

This study was supported by the National Natural Science Foundation of China (Nos. 81473401, 81173636), No. [2015]9 Guangdong province sail plan project of high level talents in 2014, and the Major science and technology projects of Guangdong province, China (No. 2013A022100039). The authors thank Dr. Haiyang He for his gift of rotavirus and technical help. The authors are also grateful to the colleagues of the SinoAmerican Cancer Research Institute, Guangdong Medical University for their useful comments on the earlier version of this manuscript.

Author’s Contribution

WCZ and LJS conceived the study; HLC, LJS, and WFC performed the research; HLC, GXL, SMZ, and RXZ analyzed and interpreted data; and HLC, XYS, and ZYP wrote the manuscript.

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    Authors and Affiliations

    1. School of Pharmacy, Guangdong Medical University, No. 1, Xincheng Road of Songshan Lake Science and Technology Industry Park, Dongguan, 523808, Guangdong Province, China

      Honglang Chen, Lijun Song, Wenfeng Chen, Shumin Zhao, Ruoxia Zhou, Xiaoying Shi, Zhenying Peng & Wenchang Zhao

    2. Institution of Traditional Chinese Medicine and New Pharmacy Development, Guangdong Medical University, Dongguan, 523808, Guangdong Province, China

      Honglang Chen, Lijun Song, Wenfeng Chen, Shumin Zhao, Ruoxia Zhou, Xiaoying Shi, Zhenying Peng & Wenchang Zhao

    3. Department of Pathology, School of Basic Medicine, Guangdong Medical University, Dongguan, 523808, Guangdong Province, China

      Guixian Li

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    Chen, H., Song, L., Li, G. et al. Human rotavirus strain Wa downregulates NHE1 and NHE6 expressions in rotavirus-infected Caco-2 cells. Virus Genes 53, 367–376 (2017). https://doi.org/10.1007/s11262-017-1444-0

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