[Effect of antisense oligonucleotides on endothelin converting enzyme expression and endothlin-1 release from airway smooth muscle cells].
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.
Wen
S
S
Guangzhou Institute of Respiratory Disease, Guangzhou 510120.
Xu
J
J
Zhong
N
N
D016282:Aspartic Acid Endopeptidases / Q000235:genetics*; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D019332:Endothelin-1 / Q000096:biosynthesis*; D000072396:Endothelin-Converting Enzymes; D015870:Gene Expression; D006801:Humans; D008168:Lung / Q000166:cytology / Q000378:metabolism; D008666:Metalloendopeptidases; D009131:Muscle, Smooth, Vascular / Q000166:cytology / Q000378:metabolism*; D020319:Oligodeoxyribonucleotides, Antisense; D014162:Transfection; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
D019332:Endothelin-1; D020319:Oligodeoxyribonucleotides, Antisense; D014409:Tumor Necrosis Factor-alpha; D016282:Aspartic Acid Endopeptidases; D008666:Metalloendopeptidases; D000072396:Endothelin-Converting Enzymes