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Karyopherin Alpha 2 Promotes the Inflammatory Response in Rat Pancreatic Acinar Cells Via Facilitating NF-κB Activation

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Abstract

Background

Activation of the transcription factor NF-κB and expression of pro-inflammatory mediators have been considered as major events of acute pancreatitis (AP). Karyopherin alpha 2 (KPNA2), a member of the importin α family, reportedly modulates p65 subcellular localization.

Aim

This study aimed to investigate the expression and possible functions of KPNA2 in the AP cell and animal model, focusing on its association with NF-κB activation.

Methods

An AP cell model was established with the cerulein-stimulated AR42J and isolated rat pancreatic acinar cells. The AP rat model was induced by the intraperitoneal injection of cerulein. The secretion of TNF-α, IL-6, and LDH was detected by ELISA kits and the production of NO using nitric oxide kit. Expression of KPNA2 was measured by RT-PCR and Western blot. Expression levels of IKKα, phosphorylation of p65, and total p65 were detected by Western blot. Co-localization of KPNA2 with p65 was observed by immunofluorescence assay. To determine the biological functions of KPNA2 in cerulein-induced inflammatory response, RNA interference was employed to knockdown KPNA2 expression in AR42J and isolated pancreatic acini cells.

Results

Cerulein stimulated KPNA2 expression and IL-6, TNF-α, NO, and LDH production in rat pancreatic acinar cells. Cerulein triggered the phosphorylation and nuclear translocation of NF-κB p65 subunit, indicating the NF-κB activation. The co-localization and nuclear accumulation of KPNA2 and p65 were detected in cerulein-treated cells. Knocking down KPNA2 hindered cerulein-induced nuclear transportation of p65 and alleviated the subsequent inflammatory response in rat pancreatic acinar cells. Additionally, KPNA2 expression was significantly up-regulated in cerulein-induced AP rat model.

Conclusions

KPNA2-facilitated p65 nuclear translocation promotes NF-κB activation and inflammation in acute pancreatitis.

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Acknowledgments

This work was supported by National Basic Research Program of China (973 Program, No. 2012CB822104); National Natural Science Foundation of China (31170766, 81470806, 31500647); the Natural Science Foundation of the Jiangsu Higher Education Institutions of China (15KJA310003); the Natural Science Foundation of Jiangsu Province (BK20150408); A Project Funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD); Development Fund for Collaborative Innovation Center of Glycoscience of Shandong University.

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    Authors and Affiliations

    1. Department of Emergency, Affiliated Hospital of Nantong University, Nantong, 226001, Jiangsu, People’s Republic of China

      Yongxia Cai, Yanbo Shen, Lili Gao, Minmin Chen & Zhongwei Huang

    2. Jiangsu Province Key Laboratory for Inflammation and Molecular Drug Target, Nantong University, Nantong, 226001, Jiangsu, People’s Republic of China

      Yongxia Cai, Yanbo Shen, Lili Gao, Minmin Chen, Zhongwei Huang & Dongmei Zhang

    3. National Glycoengineering Research Center, State Key Laboratory of Microbial Technology, Shandong University, Jinan, 250100, People’s Republic of China

      Min Xiao

    4. Department of Pathogen Biology, Medical College, Nantong University, Nantong, 226001, Jiangsu, People’s Republic of China

      Dongmei Zhang

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    1. Yongxia Cai
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    Correspondence to Zhongwei Huang or Dongmei Zhang.

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    Yongxia Cai and Yanbo Shen contribute equally to this work.

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    Cai, Y., Shen, Y., Gao, L. et al. Karyopherin Alpha 2 Promotes the Inflammatory Response in Rat Pancreatic Acinar Cells Via Facilitating NF-κB Activation. Dig Dis Sci 61, 747–757 (2016). https://doi.org/10.1007/s10620-015-3948-6

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