title: Karyopherin Alpha 2 Promotes the Inflammatory Response in Rat Pancreatic Acinar Cells Via Facilitating NF-κB Activation.
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.
  
  
  OBJECTIVE
  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.
authors.affiliation: Department of Emergency, Affiliated Hospital of Nantong University, Nantong, 226001, Jiangsu, People's Republic of China. yongxiacai@163.com.|Jiangsu Province Key Laboratory for Inflammation and Molecular Drug Target, Nantong University, Nantong, 226001, Jiangsu, People's Republic of China. yongxiacai@163.com.
authors.forename: Yongxia
authors.initials: Y
authors.lastname: Cai
authors.affiliation: Department of Emergency, Affiliated Hospital of Nantong University, Nantong, 226001, Jiangsu, People's Republic of China. yanboshen123@163.com.|Jiangsu Province Key Laboratory for Inflammation and Molecular Drug Target, Nantong University, Nantong, 226001, Jiangsu, People's Republic of China. yanboshen123@163.com.
authors.forename: Yanbo
authors.initials: Y
authors.lastname: Shen
authors.affiliation: Department of Emergency, Affiliated Hospital of Nantong University, Nantong, 226001, Jiangsu, People's Republic of China. liligao11@126.com.|Jiangsu Province Key Laboratory for Inflammation and Molecular Drug Target, Nantong University, Nantong, 226001, Jiangsu, People's Republic of China. liligao11@126.com.
authors.forename: Lili
authors.initials: L
authors.lastname: Gao
authors.affiliation: Department of Emergency, Affiliated Hospital of Nantong University, Nantong, 226001, Jiangsu, People's Republic of China. minminchen12@126.com.|Jiangsu Province Key Laboratory for Inflammation and Molecular Drug Target, Nantong University, Nantong, 226001, Jiangsu, People's Republic of China. minminchen12@126.com.
authors.forename: Minmin
authors.initials: M
authors.lastname: Chen
authors.affiliation: National Glycoengineering Research Center, State Key Laboratory of Microbial Technology, Shandong University, Jinan, 250100, People's Republic of China. minxiao@sdu.edu.cn.
authors.forename: Min
authors.initials: M
authors.lastname: Xiao
authors.affiliation: Department of Emergency, Affiliated Hospital of Nantong University, Nantong, 226001, Jiangsu, People's Republic of China. zhongweihuang88@163.com.|Jiangsu Province Key Laboratory for Inflammation and Molecular Drug Target, Nantong University, Nantong, 226001, Jiangsu, People's Republic of China. zhongweihuang88@163.com.
authors.forename: Zhongwei
authors.identifier: http://orcid.org/0000-0003-2904-0519
authors.initials: Z
authors.lastname: Huang
authors.affiliation: Jiangsu Province Key Laboratory for Inflammation and Molecular Drug Target, Nantong University, Nantong, 226001, Jiangsu, People's Republic of China. zdm@ntu.edu.cn.|Department of Pathogen Biology, Medical College, Nantong University, Nantong, 226001, Jiangsu, People's Republic of China. zdm@ntu.edu.cn.
authors.forename: Dongmei
authors.initials: D
authors.lastname: Zhang
mesh_terms: D061354:Acinar Cells / Q000378:metabolism*; D000208:Acute Disease; D000818:Animals; D015153:Blotting, Western; D002460:Cell Line; D002108:Ceruletide / Q000633:toxicity; D004195:Disease Models, Animal; D004797:Enzyme-Linked Immunosorbent Assay; D005455:Fluorescent Antibody Technique; D005786:Gene Expression Regulation; D055785:Gene Knockdown Techniques; D051550:I-kappa B Kinase / Q000378:metabolism; D007249:Inflammation; D015850:Interleukin-6 / Q000378:metabolism; D045022:Lactate Dehydrogenases / Q000378:metabolism; D008297:Male; D016328:NF-kappa B / Q000235:genetics / Q000378:metabolism; D009569:Nitric Oxide / Q000378:metabolism; D010179:Pancreas / Q000166:cytology / Q000378:metabolism*; D010195:Pancreatitis / Q000139:chemically induced / Q000235:genetics* / Q000378:metabolism; D010766:Phosphorylation; D012333:RNA, Messenger / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D020133:Reverse Transcriptase Polymerase Chain Reaction; D051996:Transcription Factor RelA / Q000235:genetics* / Q000378:metabolism; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism; D015854:Up-Regulation; D028901:alpha Karyopherins / Q000235:genetics* / Q000378:metabolism
chemical_list: D015850:Interleukin-6; D016328:NF-kappa B; D012333:RNA, Messenger; C498145:Rela protein, rat; D051996:Transcription Factor RelA; D014409:Tumor Necrosis Factor-alpha; D028901:alpha Karyopherins; C087978:karyopherin alpha 2; D009569:Nitric Oxide; D002108:Ceruletide; D045022:Lactate Dehydrogenases; D051550:I-kappa B Kinase
keywords: AR42J cells; Acute pancreatitis; Cerulein; Inflammation; Karyopherin alpha 2 (KPNA2); NF-κB