Role of ROS-induced DNA damage in high salt diet-induced vascular dysfunction

Project outline
Hypertension is a major risk factor for the development of cardiovascular diseases. Many individuals with essential hypertension are salt‐sensitive [1,2]. The exact mechanisms underlying salt-sensitive hypertension remain elusive. Preliminary data have demonstrated that in in vivo, a high salt diet (HSD) leads to vascular hypercontractility and increased blood pressure. In vitro, high sodium medium behaves as a pro-oxidant agent in vascular smooth muscle cells. Increased levels of reactive oxygen species are associated with DNA damage, which leads to the activation of PARP for DNA repair. This process leads to ADPR release, which activates TRPM2 calcium channel with the consequent increase in calcium influx [3,4].

Project aim
In this project, we aim to investigate whether vascular smooth muscle cells (VSMC) isolated from mesenteric arteries of C57BL/6 mice stimulated with high salt media (HS; 180 mM) present increased ROS-induced DNA damage when compared to cells stimulated with control media (145 mM of NaCl).

Methods
8-OHdG has been used widely in many studies as a biomarker for the measurement of endogenous oxidative DNA damage [5], and it was used to measure ROS-induced DNA damage in VSMC isolated from C57BL/6 mice treated with control or HS media. Therefore, students will analyse via ImageJ® VSMC slides stained with 8-OHdG.