Transfection of primary myometrial cells with siRNA using SilenceMag

Primary myoterial cells at 50 % confluence were transfected with 100 nM siRNA using SilenceMag.

This article highlights the capacity of Magnetofection technology to efficiently silence gene epxression into primary myometrial cells with siRNA using SilenceMag from OZ Biosciences.

article reference: Biol Reprod. 2013 Jun 5.

NOD1 and NOD2 Regulate Proinflammatory and Prolabor Mediators in Human Fetal Membranes and Myometrium via Nuclear Factor-Kappa B.
Lappas M.

Abstract

Preterm birth remains one of the most important issues facing perinatal medicine today, with chronic inflammation and/or infection being the biggest aetiological factor. The nucleotide oligomerization domain (NOD) intracellular molecules recognise a wide range of microbial products as well as other intracellular danger signals, thereby initiating inflammation through activation of nuclear factor KB (NFKB), a central regulator of the terminal processes of human labor and delivery. The aims of this study were to determine the effect of (1) human labor, pro-inflammatory cytokines and bacterial endotoxin LPS on NOD1 and NOD2 expression; and (2) NOD1 and NOD2 activation on the expression of pro-labor mediators in human fetal membranes and myometrium. NOD1 and NOD2 expression was significantly higher in fetal membranes and myometrium after spontaneous labor when compared to non-laboring tissues. Bacterial endotoxin LPS and the pro-inflammatory cytokines TNF and IL1B significantly increased NOD2, but not NOD1, expression. Furthermore, LPS-induced NOD2 expression was decreased by the NFKB inhibitor BAY 11-7082. In both fetal membranes and myometrium, the NOD1 ligand bacterial iE-DAP and the NOD2 ligand bacterial MDP significantly increased the expression and secretion of pro-inflammatory cytokines (IL6 and IL8), cyclooxygenase (PTGS2) expression and subsequent release of prostaglandins PGE₂ and PGF_2alpha, and the expression and activity of MMP9. The effects of these NOD1 and NOD2 ligands were mediated via NFKB as (i) both iE-DAP and MDP significantly increased NFKB activation, and (ii) the NFKB inhibitor BAY 11-7082 attenuated iE-DAP and MDP induced expression and secretion of pro-labor mediators. In conclusion, NOD1 and NOD2 are increased in laboring fetal membranes and myometrium and with bacterial infection. Agonist activation of NOD1 and NOD2 by bacterial products leads to NFKB activation and transcription of NFKB induced pro-labor genes. NOD1 and NOD2 may thus represent therapeutic targets for the treatment and/or management of preterm birth.

SilenceMag from OZ Biosciences is a very efficient siRNA delivery reagent based on Magnetofection™ technology.