Effects of RNA interference-induced tryptase down-regulation in P815 cells on IL-6 and TNF-[alpha] release of endothelial cells

• Published in: Journal of Zhejiang University. B. Science Vol. 9; no. 8; p. 656
• Main Authors: Jiang, Yi-feng, Zhao, Feng-di, Li, Xiao-bo, Ning, Yan-xia, Zhi, Xiu-ling, Qian, Rui-zhe, Yin, Lian-hua
• Format: Journal Article
• Language: English
• Published: Hangzhou Springer Nature B.V 01.08.2008
• Subjects: Cells; Proteins
• ISSN: 1673-1581; 1862-1783
• DOI: 10.1631/jzus.B0810188

To 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-α) of vascular endothelial cells. Tryptase-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-α 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. IL-6 and TNF-α 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-α protein levels in the medium of bEnd.3 of RNAi-P815 group were lower than those of P815 group. Reduced tryptase expression significantly inhibited the synthesis and release of IL-6 and TNF-α in vascular endothelial cells. RNA interference targeting tryptase expression may be a new anti-inflammatory strategy for vascular diseases.[PUBLICATION ABSTRACT]