Objective To evaluate the effect of a vasoactive substance
urotensin Ⅱ on the expression of type Ⅰ
collagen and migration of
human skin fibroblasts,and to explore the underlying mechanisms of
signal transduction.
Methods Fibroblasts were isolated from
human foreskin tissues and subjected to primary
culture.After a series of subculture,
fibroblasts were classified into several groups to be treated with different concentrations (10-10 to 10-6 mol/L) of
urotensin lⅡ for 24 hours,
urotensin Ⅱ of 10-s mol/L for different durations (0,4,12,24 hours),or pretreated with PD98059 (a
mitogen-activated protein kinase kinase inhibitor),
nicardipine (a
calcium channel blocker) and
ciclosporin (a
calcineurin inhibitor) of 10-5 mol/L respectively for 30 minutes followed by
treatment with
urotensin Ⅱ of 10-8 mol/L for 24 hours.The
cells receiving no
treatment served as the control.Subsequently,
enzyme-linked immunosorbent assay was performed to determine the level of
urotensin Ⅱ in the supernatant of
fibroblasts,and Transwell assay to estimate the migration activity of
fibroblasts.
Statistical analysis was carried out by t test and
analysis of variance.Results
Urotensin Ⅱ promoted the expression of type Ⅰ
collagen in a
time-and concentrationdependent manner.The level of type Ⅰ
collagen was increased by 21.2% (P > 0.05),52.2% (P < 0.05),84.4% (P <0.05),83.6% (P < 0.05) and 77.1% (P < 0.05) in the supernatant of
fibroblasts treated with 10-10,10-9,10-8,10-7 and 10-6 mol/L of
urotensin Ⅱ for 24 hours respectively,by 23.2% (P > 0.05),69.5% (P < 0.05) and 84.1% (P <0.05) in the supernatant of
fibroblasts treated with
urotensin Ⅱ of 10-8 mol/L for 4,12 and 24 hours respectively,compared with the untreated control
fibroblasts.The migration activity was markedly enhanced in
fibroblasts treated with
urotensin Ⅱ of 10-8 mol/L for 24 hours compared with the control
fibroblasts (P < 0.05).PD98059,
nicardipine and
cyclosporin A inhibited the
secretion of type Ⅰ
collagen by 18.2%,15.9% and 19.7% respectively,and suppressed the migration of
fibroblasts by 38.3% (P < 0.05),20.7% (P < 0.05) and 81.4% (P < 0.05) respectively in the groups receiving pretreatment compared with those treated with
urotensin Ⅱ alone.Conclusions
Urotensin Ⅱ can promote the
secretion of type Ⅰ
collagen by and migration of
fibroblasts,which may be realized through the Ca2+,
calmodulin kinase,and
mitogen-activated protein kinase pathways.