ABSTRACT
OBJECTIVE To investigate the beneficial effect of berberine(BBR) on atherosclerosisin Apo-/-E mice and explore the underlying mechanisms based on attenuating vascular inflammation and modulating calcification in human umbilical vein endothelial cells (HUVECs) and smooth muscle cells (SMCs). METHODS 48 Apo-/-E mice,at 6-8 weeks old,were randomly allocated into 4 groups: normal, model, bbr and atorvastatin (positive control) groups with 12 mice in each group. They were fed with high-fat diet for 4 weeks except those inNormal group and then treated with indicated drugs orsolvent for another 4 weeks. The morphology and inflammation infiltration of aortic were examined with HE staining. The expression of BMP-2 in aortic was examined by immumohistochemical staining. Blood lipid levels were examined by automatic biochemical analyzer. The expression of IL-6, TNF-α and BMP-2 in serum and tissues was detected by ELISA method. The expression of ALP and the content of calcium were detected by commercially-available kits. HUVEC cells were stimulated with TNF-α and incubated with various concentrations of BBR for 24 h. The contents of intercellular cell adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule (VCAM-1), matrix metalloprotein-9(MMP-9) in the culture supernatant were detected by ELISA method. Calcification was induced with β-glycerophosphatein SMC cells and the effect of BBR on the content of calcium was examined. RESULTS 4-week berberine treatment markedly lowered serum TC and LDL-c levels and improved the plaque stability in Apo-/-E mice fed with a high-fat diet(P<0.05 or P<0.01) which was comparable with the effect of atorvastatin. Berberineal so significantly decreased the levels of IL-6 and TNF-α in mice serum and aortic tissues (P<0.05 or P<0.001). Berberine tended to decrease ALP, BMP-2 levels and the content of calcium in mice serum and aortic tissues(P<0.05, P<0.01or P<0.001) which were not observed in atorvastatin group. Berberine significantly lowered the levels of ICAM-1, VCAM-1, and MMP-9 in TNF-α-stimulated HUVECs. It can also lowered the content of calcium in SMCs. CONCLUSION BBR can profitably regulate the levels of blood lipid in mice fed with a high-fat diet, decrease the injury caused by inflammation,and attenuate vascular calcification. It may improve atherosclerosis and play a role in cardiovascular protection.
ABSTRACT
OBJECTIVE To investigate the beneficial effect of berberine (BBR) on atherosclerosis in Apo-/-E mice and explore the underlying mechanisms based on attenuating vascular inflammation and modulating calcification in human umbilical vein endothelial cells (HUVECs) and smooth muscle cells(SMCs). METHODS 48 Apo-/-E mice,at 6-8 weeks old,were randomly allocated into 4 groups:normal, model, bbr and atorvastatin (positive control) groups with 12 mice in each group. They were fed with high-fat diet for 4 weeks except those in Normal group and then treated with indicated drugs or solvent for another 4 weeks. The morphology and inflammation infiltration of aortic were examined with HE staining. The expression of BMP- 2 in aortic was examined by immumohistochemical staining. Blood lipid levels were examined by automatic biochemical analyzer. The expression of IL-6, TNF-αand BMP-2 in serum and tissues was detected by ELISA method. The expression of ALP and the content of calcium were detected by commercially-available kits. HUVEC cells were stimulated with TNF-α and incubated with various concentrations of BBR for 24 h. The contents of intercellular cell adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule (VCAM-1), matrix metalloprotein-9 (MMP-9) in the culture supernatant were detected by ELISA method. Calcification was induced with β-glycerophos?phate in SMC cells and the effect of BBR on the content of calcium was examined. RESULTS 4-week berberine treatment markedly lowered serum TC and LDL-c levels and improved the plaquestabilityin Apo-/- E mice fed with a high- fat diet (P<0.05 or P<0.01) which was comparable with the effect of atorvastatin. Berberine also significantly decreased the levels of IL-6 and TNF-α in mice serum and aortic tissues (P<0.05 or P<0.001). Berberine tended to decrease ALP, BMP-2 levels and the content of calcium in mice serum and aortic tissues (P<0.05, P<0.01 or P<0.001) which were not observed in atorvastatin group. Berberine significantly lowered the levels of ICAM-1, VCAM-1, and MMP-9 in TNF-α-stimulated HUVECs. It can also lowered the content of calcium in SMCs. CONCLUSION BBR can profitably regulate the levels of blood lipid in mice fed with a high-fat diet, decrease the injury caused by inflammation, and attenuate vascular calcification. It may improve atherosclerosis and play a role in cardiovascular protection.