The underlying mechanism of modified palmatine for calcified cells

Abstract:

To explore the role of palmatine in vascular calcification，we employed a calcific cell model using β-glyceophosphate to stimulate the rat aortic smooth muscle cells (RASMCs). We performed a Von Kossa staining assay to detect the effect of palmatine on calcific RASMCs upon calcium deposition, we found that the modified palmatine could significantly inhibit intracellular calcium deposition of RASMCs after calcification. The mRNA level of the nuclear transcription factor nuclear factor-erythroid 2-related factor 2 (Nrf-2) was determined by Real-time Quantitative PCR, we found that the modified palmatine could significantly increase the mRNA level of Nrf-2 from (0.09±0.01) fold to (2.54±0.35) fold in RASMCs following calcification. Further，we measured the protein level of Nrf-2 by Western Blot, we found that the modified palmatine could significantly activate the expression of Nrf-2 in RASMCs following calcification. We used a Reactive Oxygen Species (ROS) kit to detect the fluorescence intensity of ROS, we observed that the modified palmatine could respectively decrease the fluorescence intensity indicating ROS from (1.04±0.03) to (0.82±0.01) in RASMCs after calcification. We used a Nitric Oxide (NO) kit to measure the production of NO, we observed that the modified palmatine could decrease the production of NO from (5.73±0.05) μmol/L to (3.73±0.15) μmol/L in RASMCs after calcification.