Primary amnion cells and myometrial cells at approximately 50% confluence were transfected using SilenceMag reagent and 50 nM siRNA for 48H.
This article highlights the capacity of Magnetofection technology to efficiently silence gene epxression into human primary myometrial cells with siRNA using SilenceMag from OZ Biosciences.article reference: Am J Reprod Immunol. 2013 Nov 29. doi: 10.1111/aji.12181.
SLIT3 is Increased in Supracervical Human Foetal Membranes and in Labouring Myometrium and Regulates Pro-Inflammatory Mediators.
Lim R, Barker G, Lappas M.Abstract
PROBLEM: Inflammation is associated with preterm birth, a worldwide healthcare issue. SLIT3 has a role in inflammation, and thus, the purpose of this study was to determine the effect of SLIT3 on labour mediators in human gestational tissues.
METHOD OF STUDY: SLIT3 protein expression was performed using immunohistochemistry in foetal membranes and myometrium with no labour and after labour. Foetal membranes were also obtained from a distal site (DS) and supracervical site (overlying the cervix; SCS). SLIT3 gene silencing was achieved using siRNA in primary amnion and myometrial cells. Pro-inflammatory and pro-labour mediators were evaluated by qRT-PCR, ELISA and gelatin zymography.
RESULTS: SLIT3 expression was greater in foetal membranes from the SCS compared with DS and in myometrium after term spontaneous labour onset. SLIT3 siRNA in primary amnion and myometrial cells decreased IL-1β-induced pro-inflammatory cytokine gene expression and release (IL-6 and IL-8) and MMP-9 gene expression and release. In amnion cells, SLIT3 siRNA knockdown decreased IL-1β-induced COX-2 expression and prostaglandin PGE2 release. There was no effect of SLIT3 siRNA on IL-1β-induced NF-κB transcriptional activity.
CONCLUSION: Our results demonstrate that SLIT3 is increased with labour, and both our amnion and our myometrial studies describe a pro-inflammatory effect of SLIT3 in these tissues.
No comments:
Post a Comment