Inhibition of the K+ channel K(Ca)3.1 reduces TGF-β1-induced premature senescence, myofibroblast phenotype transition and proliferation of mesangial cells.
OBJECTIVE
K(Ca)3.1 channel participates in many important cellular functions. This study planned to investigate the potential involvement of K(Ca)3.1 channel in premature senescence, myofibroblast phenotype transition and proliferation of mesangial cells.


METHODS
Rat mesangial cells were cultured together with TGF-β1 (2 ng/ml) and TGF-β1 (2 ng/ml) + TRAM-34 (16 nM) separately for specified times from 0 min to 60 min. The cells without treatment served as controls. The location of K(Ca)3.1 channels in mesangial cells was determined with Confocal laser microscope, the cell cycle of mesangial cells was assessed with flow cytometry, the protein and mRNA expression of K(Ca)3.1, α-smooth muscle actin (α-SMA) and fibroblast-specific protein-1 (FSP-1) were detected with Western blot and RT-PCR. One-way analysis of variance (ANOVA) and Student-Newman-Keuls-q test (SNK-q) were used to do statistical analysis. Statistical significance was considered at P<0.05.


RESULTS
Kca3.1 channels were located in the cell membranes and/or in the cytoplasm of mesangial cells. The percentage of cells in G0-G1 phase and the expression of K(ca)3.1, α-SMA and FSP-1 were elevated under the induction of TGF-β1 when compared to the control and decreased under the induction of TGF-β1+TRAM-34 when compared to the TGF-β1 induced (P<0.05 or P<0.01).


CONCLUSIONS
Targeted disruption of K(Ca)3.1 inhibits TGF-β1-induced premature aging, myofibroblast-like phenotype transdifferentiation and proliferation of mesangial cells.
Department of Nephrology, Second Affiliated Hospital, School of Medicine, Xi'an Jiaotong University, Xi'an, Shaanxi Province, P.R. China.
Rong-Guo
RG
Fu
School of Medicine, Xi'an Jiaotong University, Xi'an, Shaanxi Province, P.R. China.
Tao
T
Zhang
Department of Nephrology, Second Affiliated Hospital, School of Medicine, Xi'an Jiaotong University, Xi'an, Shaanxi Province, P.R. China.
Li
L
Wang
Department of Nephrology, Second Affiliated Hospital, School of Medicine, Xi'an Jiaotong University, Xi'an, Shaanxi Province, P.R. China.
Yan
Y
Du
Department of Nephrology, Second Affiliated Hospital, School of Medicine, Xi'an Jiaotong University, Xi'an, Shaanxi Province, P.R. China.
Li-Ning
LN
Jia
School of Medicine, Xi'an Jiaotong University, Xi'an, Shaanxi Province, P.R. China.
Jing-Jing
JJ
Hou
Department of Nephrology, Second Affiliated Hospital, School of Medicine, Xi'an Jiaotong University, Xi'an, Shaanxi Province, P.R. China.
Gang-Lian
GL
Yao
Cadre's ward, Second Affiliated Hospital, School of Medicine, Xi'an Jiaotong University, Xi'an, P.R. China.
Xiao-Dan
XD
Liu
Medical Laboratory, Second Affiliated Hospital, School of Medicine, Xi'an Jiaotong University, Xi'an, P.R. China.
Lei
L
Zhang
Department of Oncology, First Affiliated Hospital, School of Medicine, Xi'an Jiaotong University, Xi'an, Shaanxi Province, P.R. China.
Ling
L
Chen
Department of Nephrology, Second Affiliated Hospital, School of Medicine, Xi'an Jiaotong University, Xi'an, Shaanxi Province, P.R. China.
Bao-Song
BS
Gui
Department of Anesthesia, Second Affiliated Hospital, School of Medicine, Xi'an Jiaotong University, Xi'an, Shaanxi Province, P.R. China.
Rong-Liang
RL
Xue
D000818:Animals; D018929:Cell Culture Techniques; D002453:Cell Cycle; D002462:Cell Membrane / Q000378:metabolism; D049109:Cell Proliferation; D002478:Cells, Cultured; D016922:Cellular Senescence; D051660:Intermediate-Conductance Calcium-Activated Potassium Channels / Q000037:antagonists & inhibitors*; D050527:Mesangial Cells / Q000166:cytology* / Q000378:metabolism; D058628:Myofibroblasts / Q000166:cytology* / Q000378:metabolism; D010641:Phenotype; D011720:Pyrazoles / Q000494:pharmacology; D051381:Rats; D053773:Transforming Growth Factor beta1 / Q000378:metabolism / Q000494:pharmacology
D051660:Intermediate-Conductance Calcium-Activated Potassium Channels; C491575:Kcnn4 protein, rat; D011720:Pyrazoles; C411671:TRAM 34; D053773:Transforming Growth Factor beta1