Gene Silencing in Human Coronary Artery Endothelial Cells (HCAEC) with siRNA using SilenceMag (Magnetofection)

siRNA (10 nM) were incubated with SilenceMag according to the recommendation protocol and added onto HCAEC seeded into 6 well plates. 

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

article reference: Cardiovasc Diabetol. 2014 Nov 13;13(1):152.

Activation of AMP-activated protein kinase by metformin protects human coronary artery endothelial cells against diabetic lipoapoptosis.

Eriksson L, Nyström T.


abstract
BackgroundThe prevalence of type 2 diabetes (T2D) among adults worldwide is rapidly increasing, and in patients with diabetes the major cause of death is macrovascular disease. Endothelial cells play an important role in maintaining vascular homeostasis. Free fatty acids, which are elevated in T2D, have previously been shown to induce endothelial dysfunction and apoptosis of endothelial cells, which is considered as an important and early factor in the onset of atherosclerosis and cardiovascular disease. Metformin, which is used as first line treatment of T2D patients, is believed to exert its pharmacological effects through activation of AMP-activated protein kinase, which has emerged as a new potential target in reversing endothelial dysfunction.MethodsHere we studied the protective effect of metformin against free fatty acid-induced apoptosis of human coronary artery endothelial cells (HCAECs) by assessing DNA fragmentation and cleaved caspase 3 levels. We also attempted to elucidate the underlying mechanisms by investigating the involvement of AMP-activated protein kinase, p38 MAPK and eNOS. Generation of reactive oxygen species by free fatty acid exposure was also examined.ResultsOur results suggest that metformin protects HCAECs from lipoapoptosis, an effect that involves eNOS and p38 MAPK, downstream of AMPK signaling, but not as previously suggested through suppression of reactive oxygen species.ConclusionThe protective effect of metformin against free fatty acid induced apoptosis is potentially clinically relevant as metformin is first line treatment for patients with T2D, a patient group which is rapidly increasing and carries a high burden of cardiovascular disease.

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