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[Effect of antisense oligonucleotides on endothelin converting enzyme expression and endothlin-1 release from airway
Objective:
To explore the mechanism of TNF-alpha induced ET-1 release and evaluate the effects of endothelin converting enzyme (ECE) antisense oligonucleotides (oDNS) on TNF-alpha induced ET-1 release from airway smooth muscle cells (ASMC).
Methods:
ASMC transfected with the antisense oligonucleodides of ECE vectored with lipofectin were established. TNF-alpha induced ECE-mRNA expression and ET-1 release from above cultured cells were determined by RT-PCR and ELISA.
Results:
ET-1 levels (10.1 +/- 0.3) ng/L and ECE expression (ECE/beta-actin: 0.228 +/- 0.015) in cultured ASMC incubated with 1,000 U/ml TNF-alpha were significantly higher than those in controls without TNF-alpha incubation (7.3 +/- 0.8) ng/L, (0.132 +/- 0.032, all P < 0.01). However, after transfecting antisence ECE-oDNS in those cells, ET-1 levels and ECE expressions were significantly lower than those in control cells without antisense transfection. The inhibition rates were (29 +/- 5)% and (35 +/- 12)% respectively.
Conclusions:
Abnormal expression of ECE mRNA may be a keypoint responsible for TNF-alpha induced ET-1 release in human ASMC; TNF-alpha induced ET-1 release and ECE mRNA expression from human ASMC are inhibited by antisense oligonucleotide of ECE.