[The effects of endothelin-1 and selective endothelin receptor-type A antagonist on human renal interstitial fibroblasts in vitro].
OBJECTIVE
(1) To investigate the effects of endothelin-1 (ET-1) on cell proliferation of cultured human renal interstitial fibroblasts (hRIFs), and the mRNA expression of type I Collagen (Col I), transforming growth factor-beta (TGF-beta), matrix metalloproteinase-1 (MMP-1) and tissue inhibitors of metalloproteinase-1, -2 (TIMP-1, TIMP-2) by hRIFs. (2) To investigate the changes of above effects after the hRIFs were pretreated with selective endothelin receptor-type A antagonist (ETaRA).


METHODS
(1) The proliferation of hRIFs was determined by MTT method. (2) The mRNA expression of Col I, TGF-beta, MMP-1, TIMP-1 and TIMP-2 was detected semiquantitatively with reverse transcription-polymerase chain reaction (RT-PCR).


RESULTS
(1) The hRIFs proliferation was increased by ET-1 stimulation in dose dependent manner (10(-11) approximately 10(-7) mol/L, 24 h), and the peak growth level was at the concentration of 10(-7) mol/L (P < 0.05). (2) The hRIFs proliferation was significantly increased at 8th hour after ET-1 stimulation (10(-7) mol/L) (P < 0.01), and the peak growth level was attained at 24th hour (P < 0.01). (3) The mRNA expression of Col I, TGF-beta, MMP-1, TIMP-1 and TIMP-2 by hRIFs was upregulated with ET-1 in dose dependent manner (10(-11) approximately 10(-7) mol/L, 16 h), and the peak expression level was at the concentration of 10(-7) mol/L (P < 0.05). (4) When hRIFs were stimulated by ET-1 (10(-7) mol/L), the mRNA expression of Col I and TGF-beta was significantly increased at 8th hour (P < 0.05), and the peak expression levels were at 24th hour and 8th hour respectively; the mRNA expression of TIMP-1 and TIMP-2 was significantly increased at 16th hour (P < 0.05) and reached the peak level at 24th hour; the mRNA expression of MMP-1 was significantly increased and attained the peak level at 16th hour (P < 0.05). (5) The above effects of ET-1 were significantly inhibited by ETaRA (P < 0.05).


CONCLUSIONS
The stimulating effects of ET-1 on hRIFs proliferation and the mRNA expression of Col I, TGF-beta, MMP-1, TIMP-1 and TIMP-2, and the inhibiting effects of ETaRA on the above effects suggest that ET-1 may participate in the process of renal interstitial fibrosis under pathological condition.
Tian
Xuefei
X
Division of Nephrology, China-Japan Friendship Hospital, Beijing 100029, China.
Tang
Gongyao
G
Chen
Yipu
Y
D002455:Cell Division / Q000187:drug effects; D024042:Collagen Type I / Q000235:genetics / Q000378:metabolism; D065128:Endothelin Receptor Antagonists; D019332:Endothelin-1 / Q000494:pharmacology*; D005347:Fibroblasts / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D005355:Fibrosis / Q000378:metabolism / Q000473:pathology; D006801:Humans; D007668:Kidney / Q000187:drug effects / Q000473:pathology; D020781:Matrix Metalloproteinase 1 / Q000235:genetics / Q000378:metabolism; D009842:Oligopeptides / Q000494:pharmacology*; D012333:RNA, Messenger / Q000187:drug effects / Q000378:metabolism; D044022:Receptor, Endothelin A; D017466:Receptors, Endothelin / Q000378:metabolism; D019715:Tissue Inhibitor of Metalloproteinase-1 / Q000235:genetics / Q000378:metabolism; D019716:Tissue Inhibitor of Metalloproteinase-2 / Q000235:genetics / Q000378:metabolism; D016212:Transforming Growth Factor beta / Q000235:genetics / Q000378:metabolism
D024042:Collagen Type I; D065128:Endothelin Receptor Antagonists; D019332:Endothelin-1; D009842:Oligopeptides; D012333:RNA, Messenger; D044022:Receptor, Endothelin A; D017466:Receptors, Endothelin; D019715:Tissue Inhibitor of Metalloproteinase-1; D016212:Transforming Growth Factor beta; C412729:acetyl-tryptophyl-leucyl-aspartyl-isoleucyl-isoleucyl-tryptophan; D019716:Tissue Inhibitor of Metalloproteinase-2; D020781:Matrix Metalloproteinase 1