PMID- 24291552
OWN - NLM
STAT- MEDLINE
DCOM- 20141106
LR  - 20180815
IS  - 1878-5875 (Electronic)
IS  - 1357-2725 (Linking)
VI  - 47
DP  - 2014 Feb
TI  - Inhibition of KCa3.1 suppresses TGF-beta1 induced MCP-1 expression in human proximal 
      tubular cells through Smad3, p38 and ERK1/2 signaling pathways.
PG  - 1-10
LID - S1357-2725(13)00355-5 [pii]
LID - 10.1016/j.biocel.2013.11.017 [doi]
AB  - It is well known that TGF-beta1 plays a central role in renal fibrosis due in large 
      part to stimulation of inflammatory responses. KCa3.1, a potassium channel 
      protein, has been suggested as a potential therapeutic target for diseases such 
      as sickle cell anemia, autoimmunity, atherosclerosis and more recently, kidney 
      fibrosis. Blockade of KCa3.1 has been shown to ameliorate renal fibrosis in 
      diabetic mice in association with reduced TGF-beta1 signaling. However, the 
      centrality of KCa3.1 activation to TGF-beta1 induced inflammation remains unknown. 
      In this study, human proximal tubular cells (HK2 cells) were incubated with 
      TGF-beta1 (2 ng/ml) for 48 h in the presence or absence of KCa3.1 siRNA or the 
      KCa3.1 inhibitor TRAM34. HK2 cells overexpressing KCa3.1 were studied in 
      parallel. The mRNA and protein expression of monocyte chemoattractant protein-1 
      (MCP-1) were measured by qRT-PCR and ELISA. Downstream TGF-beta1 signaling molecules 
      Smad3, p38 and ERK1/2 were measured by Western blot analysis. Using whole-cell 
      patch clamp techniques we found that TGFbeta-1 induced a large KCa3.1 K-current that 
      was inhibited by TRAM34. TGF-beta1 also increased MCP-1 mRNA and protein expression 
      in HK2 cells compared to control, an effect that was reversed by in the presence 
      of KCa3.1 siRNA. Similarly, TRAM34 significantly reduced the TGF-beta1-mediated 
      increase in MCP-1 at both the mRNA and protein levels. Inhibition of KCa3.1 with 
      KCa3.1 siRNA or TRAM34 also reduced TGF-beta1-induced phosphorylation of Smad3, p38 
      and ERK1/2 MAPK pathways. Conversely overexpression of KCa3.1 induced TGF-beta1 
      signaling cascades and expression of MCP-1. The present study is consistent with 
      a key role for KCa3.1 renal proximal tubular cells in mediating the TGF-beta1 
      induction of MCP-1 expression in HK2 cells via Smad3, p38 and ERK1/2 MAPK 
      signaling pathways.
CI  - Copyright (c) 2013 Elsevier Ltd. All rights reserved.
FAU - Huang, Chunling
AU  - Huang C
AD  - Kolling Institute of Medical Research, Sydney Medical School, University of 
      Sydney, Royal North Shore Hospital, St Leonards, NSW 2065, Australia; Xiamen 
      Center of Clinical Laboratory, Xiamen Zhongshan Hospital, Medical College of 
      Xiamen University, Xiamen 361004, China.
FAU - Day, Margot L
AU  - Day ML
AD  - School of Medical Sciences, Discipline of Physiology and Bosch Institute, 
      University of Sydney, Sydney, NSW 2006, Australia.
FAU - Poronnik, Philip
AU  - Poronnik P
AD  - School of Medical Sciences, Discipline of Physiology and Bosch Institute, 
      University of Sydney, Sydney, NSW 2006, Australia.
FAU - Pollock, Carol A
AU  - Pollock CA
AD  - Kolling Institute of Medical Research, Sydney Medical School, University of 
      Sydney, Royal North Shore Hospital, St Leonards, NSW 2065, Australia. Electronic 
      address: carol.pollock@sydney.edu.au.
FAU - Chen, Xin-Ming
AU  - Chen XM
AD  - Kolling Institute of Medical Research, Sydney Medical School, University of 
      Sydney, Royal North Shore Hospital, St Leonards, NSW 2065, Australia.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20131128
PL  - Netherlands
TA  - Int J Biochem Cell Biol
JT  - The international journal of biochemistry & cell biology
JID - 9508482
RN  - 0 (CCL2 protein, human)
RN  - 0 (Chemokine CCL2)
RN  - 0 (Flavonoids)
RN  - 0 (Imidazoles)
RN  - 0 (Intermediate-Conductance Calcium-Activated Potassium Channels)
RN  - 0 (KCNN4 protein, human)
RN  - 0 (Pyrazoles)
RN  - 0 (Pyridines)
RN  - 0 (RNA, Small Interfering)
RN  - 0 (SMAD3 protein, human)
RN  - 0 (Smad3 Protein)
RN  - 0 (TGFB1 protein, human)
RN  - 0 (TRAM 34)
RN  - 0 (Transforming Growth Factor beta1)
RN  - EC 2.7.11.24 (p38 Mitogen-Activated Protein Kinases)
RN  - OU13V1EYWQ (SB 203580)
RN  - SJE1IO5E3I (2-(2-amino-3-methoxyphenyl)-4H-1-benzopyran-4-one)
SB  - IM
MH  - Cells, Cultured
MH  - Chemokine CCL2/*biosynthesis/genetics/metabolism
MH  - Flavonoids/pharmacology
MH  - Gene Knockdown Techniques
MH  - Humans
MH  - Imidazoles/pharmacology
MH  - Intermediate-Conductance Calcium-Activated Potassium Channels/*antagonists & 
      inhibitors/genetics/metabolism
MH  - Kidney Tubules, Proximal/enzymology/metabolism
MH  - MAP Kinase Signaling System/drug effects
MH  - Phosphorylation
MH  - Pyrazoles/pharmacology
MH  - Pyridines/pharmacology
MH  - RNA, Small Interfering/administration & dosage/genetics
MH  - Signal Transduction
MH  - Smad3 Protein/genetics/*metabolism
MH  - Transfection
MH  - Transforming Growth Factor beta1/*antagonists & inhibitors/genetics/metabolism
MH  - p38 Mitogen-Activated Protein Kinases/*metabolism
OTO - NOTNLM
OT  - Cell signaling
OT  - KCa3.1
OT  - MCP-1
OT  - TGF-beta1
EDAT- 2013/12/03 06:00
MHDA- 2014/11/07 06:00
CRDT- 2013/12/03 06:00
PHST- 2013/07/26 00:00 [received]
PHST- 2013/11/13 00:00 [revised]
PHST- 2013/11/18 00:00 [accepted]
PHST- 2013/12/03 06:00 [entrez]
PHST- 2013/12/03 06:00 [pubmed]
PHST- 2014/11/07 06:00 [medline]
AID - S1357-2725(13)00355-5 [pii]
AID - 10.1016/j.biocel.2013.11.017 [doi]
PST - ppublish
SO  - Int J Biochem Cell Biol. 2014 Feb;47:1-10. doi: 10.1016/j.biocel.2013.11.017. 
      Epub 2013 Nov 28.