Inhibitory effects of lithospermic acid on proliferation and migration of rat vascular smooth muscle cells

Abstract

Aim: To understand the effects of lithospermic acid (LA), a potent antioxidant from the water-soluble extract of Salvia miltiorrhiza, on the migration and proliferation of rat thoracic aorta vascular smooth muscle cells (VSMCs). Methods: VSMC migration, proliferation, DNA synthesis and cell cycle progression were investigated by transwell migration analysis, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, bromodeoxyuridine (BrdU) incorporation assay, and flow cytometric detection, respectively. Intracellular reactive oxygen species (ROS) generation was detected using 2′,7′-dichlorofluorescin diacetate (DCFH-DA). The expression of cyclin D1 protein and matrix metalloproteinase-9 (MMP-9) protein, as well as the phosphorylation state of ERK1/2, were determined using Western blots. The activity of MMP-9 and the expression of MMP-9 mRNA were assessed by gelatin zymography analysis and RT-PCR, respectively. Results: LA (25−100 μmol/L) inhibited both lipopolysaccharide (LPS)-and fetal bovine serum (FBS)-induced ROS generation and ERK1/2 phosphorylation. By down-regulating the expression of cyclin D 1 and arresting cell cycle progression at the G 1 phase, LA inhibited both VSMC proliferation and DNA synthesis as induced by 5% FBS. Furthermore, LA attenuated LPS-induced VSMC migration by inhibiting MMP-9 expression and its enzymatic activity. Conclusion: LA is able to inhibit FBS-induced VSMC proliferation and LPS-induced VSMC migration, which suggests that LA may have therapeutic effects in the prevention of atherosclerosis, restenosis and neointimal hyperplasia.

Phenotypic Modulation of Mesenteric Vascular Smooth Muscle Cells from Type 2 Diabetic Rats is Associated with Decreased Caveolin-1 Expression

“…To investigate the effect of HG on ROS generation, quiescent mesenteric VSMCs were treated with HG or NG for 12 hours. In another set of experiments, to measure ROS generation and ERK1/2 activation, quiescent cells were stimulated with LPS (100 ng/mL) for 15 minutes [25,26]. …”

Chlorogenic acid inhibits hypoxia-induced pulmonary artery smooth muscle cells proliferation via c-Src and Shc/Grb2/ERK2 signaling pathway

“…Reactive oxygen species production was quantified by the 2 0 ,7 0 -dichlorofluorescin diacetate (DCFH-DA) method (Chen et al, 2009), based on the reactive oxygen species-dependent oxidation of DCFH-DA to 2 0 ,7 0 -dichlorofluorescein (DCF). PASMCs grown in 96-well plates were then incubated with DCFH-DA (10 μmol/l) for 30 min.…”

“…The cell proliferation assay was performed using the MTT method as described previously (Chen et al, 2009). Briefly, PASMCs were treated under normoxia or hypoxia for 48 h, and then the cells were incubated with 0.5 mg/ml MTT for 4 h at 37 1C.…”

Pharmacological actions and therapeutic applications of Salvia miltiorrhiza depside salt and its active components

“…Our group reported that LA exerted vasodilator action by modulating Ca 2+ homeostasis in VSMCs [63] and prevented atherosclerosis by inhibiting VSMC proliferation and migration [111] .…”