title: [Nuclear factor-κB pathway mediates the effects of CD137 signaling on NFATc1 expression in mice vascular smooth muscle cells].
abstract: OBJECTIVE
  To observe whether CD137 signaling could affect the nuclear factor of activated T cells c1 (NFATc1) expression through nuclear factor-κB (NF-κB) pathway in mice aortic vascular smooth muscle cells (VSMCs).
  
  
  METHODS
  Adherence methods for tissues explants were used for primary culture of mouse aortic VSMCs. The mRNA expression of CD137 and NFATc1 was detected by real-time quantitative PCR (RT-qPCR). The VSMCs protein expression of IκB-α, NF-κB p65, phospo-p65 and NFATc1 was determined by Western blot. The level of CD137 was measured by Flow Cytometry (FCM).
  
  
  RESULTS
  (1) The mRNA and protein expression of CD137 in VSMCs was significantly upregulated at 24 h after co-culture with TNF-α (10 ng/ml, all P < 0.05). (2) Compared with the control group, the level of p-NF-κB p65 in cytoplasm and nucleus was significantly increased (8.34 ± 0.28 vs. 1, P < 0.05, and 2.64 ± 0.42 vs. 1, P < 0.05) while the level of IκB-α was reduced (1 vs. 2.70 ± 0.28, P < 0.05) after co-treatment with agonist-CD137 mAb, above effects were partly blocked by adding specific NF-κB inhibitor PDTC (30 µmol/L: 1.15 ± 0.14 vs. 8.34 ± 0.28, P < 0.05, and 2.09 ± 0.12 vs. 2.64 ± 0.42, P < 0.05, and 1.78 ± 0.74 vs. 1, P < 0.05). (3) The mRNA (2.07 ± 0.09 vs. 1, P < 0.05) and protein (1.75 ± 0.07 vs. 1, P < 0.05) expression of NFATc1 was significantly upregulated by agonist CD137mAb compared with the control group, and these effects could be reversed by PDTC (1.15 ± 0.07 vs. 2.07 ± 0.09, P < 0.05, and 0.90 ± 0.11 vs. 1.75 ± 0.07, P < 0.05).
  
  
  CONCLUSIONS
  CD137 signaling could affect the NFATc1expression in VSMCs through NF-kappaB pathway.
authors.affiliation: Department of Cardiology, Affiliated Hospital of Jiangsu University, Zhenjiang 212001, China.
authors.forename: Yunjie
authors.initials: Y
authors.lastname: Yin
authors.affiliation: Department of Cardiology, Affiliated Hospital of Jiangsu University, Zhenjiang 212001, China; Email: yanjinchuan@hotmail.com.
authors.forename: Jinchuan
authors.initials: J
authors.lastname: Yan
authors.affiliation: Department of Cardiology, Affiliated Hospital of Jiangsu University, Zhenjiang 212001, China.
authors.forename: Zhongqun
authors.initials: Z
authors.lastname: Wang
authors.affiliation: Department of Cardiology, Affiliated Hospital of Jiangsu University, Zhenjiang 212001, China.
authors.forename: Peijing
authors.initials: P
authors.lastname: Liu
authors.affiliation: Department of Cardiology, Affiliated Hospital of Jiangsu University, Zhenjiang 212001, China.
authors.forename: Yi
authors.initials: Y
authors.lastname: Liang
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D020672:I-kappa B Proteins; D051379:Mice; D009131:Muscle, Smooth, Vascular / Q000378:metabolism*; D032389:Myocytes, Smooth Muscle; D000072000:NF-KappaB Inhibitor alpha; D016328:NF-kappa B / Q000378:metabolism*; D050778:NFATC Transcription Factors / Q000378:metabolism*; D012333:RNA, Messenger; D015398:Signal Transduction; D053261:Tumor Necrosis Factor Receptor Superfamily, Member 9 / Q000378:metabolism*; D014409:Tumor Necrosis Factor-alpha; D015854:Up-Regulation
chemical_list: D020672:I-kappa B Proteins; D016328:NF-kappa B; D050778:NFATC Transcription Factors; C000606205:Nfkbia protein, mouse; D012333:RNA, Messenger; D053261:Tumor Necrosis Factor Receptor Superfamily, Member 9; D014409:Tumor Necrosis Factor-alpha; D000072000:NF-KappaB Inhibitor alpha