Effects of RNA interference-induced tryptase down-regulation in P815 cells on IL-6 and TNF-alpha release of endothelial cells.
- Author:
Yi-feng JIANG
1
;
Feng-di ZHAO
;
Xiao-bo LI
;
Yan-xia NING
;
Xiu-ling ZHI
;
Rui-zhe QIAN
;
Lian-hua YIN
Author Information
- Publication Type:Journal Article
- MeSH: Animals; Cell Line; Culture Media, Conditioned; Down-Regulation; genetics; Endothelial Cells; enzymology; secretion; Gene Expression Regulation, Enzymologic; Interleukin-6; genetics; secretion; Mice; RNA, Messenger; genetics; RNA, Small Interfering; genetics; Transgenes; Tryptases; genetics; metabolism; Tumor Necrosis Factor-alpha; genetics; secretion
- From: Journal of Zhejiang University. Science. B 2008;9(8):656-661
- CountryChina
- Language:English
-
Abstract:
OBJECTIVETo explore the effects of down-regulated tryptase expression in mast cells on the synthesis and release of interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-alpha) of vascular endothelial cells.
METHODSTryptase-siRNA (small-interfering RNA) vector was constructed to inhibit tryptase expression in P815 cells. The medium of P815 cells treated by the tryptase-siRNA (RNAi-P815 group) or pure vector (P815 group) was collected and used to culture bEnd.3 cells. The messenger RNAs (mRNAs) of IL-6 and TNF-alpha in bEnd.3 cells and their protein levels in the medium were measured by reverse transcription polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay (ELISA), respectively.
RESULTSIL-6 and TNF-alpha mRNAs in bEnd.3 cells cultured in RNAi-P815-conditioned medium decreased significantly compared to those in P815-conditioned medium. Consistently, IL-6 and TNF-alpha protein levels in the medium of bEnd.3 of RNAi-P815 group were lower than those of P815 group.
CONCLUSIONReduced tryptase expression significantly inhibited the synthesis and release of IL-6 and TNF-alpha in vascular endothelial cells. RNA interference targeting tryptase expression may be a new anti-inflammatory strategy for vascular diseases.
