Lysophosphatidic acid-activated Cl- current activity in human systemic sclerosis skin fibroblasts.
OBJECTIVE
SSc (scleroderma) is an often fatal disease characterized by widespread tissue fibrosis. Fibroblasts play a key role in SSc-associated fibrosis. This study was designed to determine: (i) whether fibroblasts isolated from skin of patients with SSc have increased lysophosphatidic acid-activated Cl- current (IClLPA) activity vs healthy controls; (ii) whether myofibroblast differentiation is involved in SSc skin fibrosis; and (iii) whether SSc fibroblasts have different proliferation rates vs controls.


METHODS
Skin biopsies were taken from involved and uninvolved skin of SSc patients and controls. Whole-cell perforated patch-clamping was used to measure IClLPA activity in fibroblasts isolated and cultured from these biopsies. Western blotting was used to measure α-smooth muscle actin (α-SMA). Proliferation was measured using a colorimetric assay.


RESULTS
Fibroblasts cultured from SSc skin show significantly increased IClLPA activity following LPA exposure compared with control skin fibroblasts. α-SMA protein was significantly increased in cultured SSc skin fibroblasts vs controls. No significant differences in proliferation rates were found.


CONCLUSIONS
Elevated IClLPA activity is a hallmark of SSc skin fibroblasts. Blocking IClLPA activation may be a new therapeutic approach for treating SSc-associated fibrosis.
Yin
Zhaohong
Z
Department of Physiology, University of Tennessee Health Science Center, 894 Union Ave., Memphis, TN 38163, USA.
Carbone
Laura D
LD
Gotoh
Mari
M
Postlethwaite
Arnold
A
Bolen
Alyssa L
AL
Tigyi
Gabor J
GJ
Murakami-Murofushi
Kimiko
K
Watsky
Mitchell A
MA
D000199:Actins / Q000378:metabolism*; D000328:Adult; D000704:Analysis of Variance; D016022:Case-Control Studies; D002478:Cells, Cultured; D002712:Chlorides / Q000378:metabolism*; D005260:Female; D005347:Fibroblasts / Q000378:metabolism*; D006801:Humans; D008246:Lysophospholipids / Q000378:metabolism*; D008297:Male; D008875:Middle Aged; D009130:Muscle, Smooth / Q000378:metabolism; D058628:Myofibroblasts / Q000378:metabolism; D012595:Scleroderma, Systemic / Q000378:metabolism*; D012867:Skin / Q000378:metabolism
D000199:Actins; D002712:Chlorides; D008246:Lysophospholipids; C032881:lysophosphatidic acid
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