A salvianolic acid 8-rich fraction of Sa/via miltiorrhiza induces neointimal cell apoptosis in rabbit angioplasty model

Abstract

Apoptosis has been suggested to participate in stabilizing cell number in restenosis. Salvia miltiorrhiza (SM) Bunge which is a Chinese herb widely used for the treatment of cardiovascular disorders contains a potent antioxidant, Salvianolic ac id B. To determ ine whether the antioxidant affects vascular apoptosis, the present study examined the frequency of apoptotic cell death in atherosclerotic plaques and in restenotic lesions of cholesterol-fed rabbits. New Zealand White rabbits were treated with a normal diet (normal), a 2% cholesterol diet (HC), a 2% cholesterol diet and endothe lial denudation (HC-ED), a 2 % cholesterol diet with 5% water-soluble extract of SM (4.8 glKg B. W./day) and endothelial denudation (HCED-SM), or with a 2% cholesterol diet containing probucol (0.6 glkg B.W./day) and endothelial denudation (HC-ED-probucol). Apoptosis and associated cell types were examined in serial paraffin sections by in situ terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling and immunohistochemistry. The expression of p53, an apoptosis-related protein, was also examined. Apoptosis was mainly detected in the neointima of the three gro ups with endothe lial denudation. The percentage of apoptotic cells in SMtreated group (68.5±5.9%) was significantly higher than that of normal (0 %), HC (1.9±1.2 %), HC-ED (46.1±5.4%), and probucol-treated (32.8±3.9%) groups. The SM treatment markedly reduced the thickness of the neointima which was mainly composed of smooth muscle cells with few macrophages. In accordance with the apoptotic cell counts, positive immunoreactivity for p53 was observed in restenotic lesions from HC-ED, SM-treated and probucol-treated groups but not in the intima of the other two groups. These results suggest that the treatment with salvianolic acid B-rich fraction of SM induces apoptosis in neointima which in turn may help prevent the neointimal thickening.