Yixintongmai Inhibits Proliferation and Migration and Promotes Apoptosis of Vascular Smooth Muscle Cells Cultured with High Glucose

Abstract

Objective. This study was designed to evaluate the effects of yixintongmai on proliferation, migration, and apoptosis of vascular smooth muscle cells (VSMCs) cultured with high glucose. Methods. VSMCs of the thoracic aorta from 5- to 8-week-old male Sprague-Dawley rats were cultured with normal (4.5 mM) or high (25 mM) glucose, respectively. The concentration of yixintongmai powder at 360 μg/ml was chosen according to pre-experimental results. Results. Yixintongmai inhibited the proliferation of VSMCs (CCK-8 assay: 0.75 ± 0.04 versus 0.98 ± 0.09 OD, P < 0.001 ; cell counting: 37533 ± 1861 versus 56009 ± 3779 cells/well, P < 0.001 ) and the expression of proliferating cell nuclear antigen (0.74 ± 0.08 fold, P < 0.001 ) as compared with high glucose (HG). Yixintongmai inhibited the migration of VSMCs (transwell assay: 146 ± 16 versus 265 ± 62 cells; P < 0.001 ), scratch wound assay (0.17 ± 0.01 fold, P < 0.001 ), and the expression of matrix metalloproteinases-9 (0.87 ± 0.03 fold, P < 0.001 ) as compared with HG. Yixintongmai decreased mitochondrial membrane potentials (0.36 ± 0.12 fold, P < 0.001 ) and promoted early (2.11 ± 0.20 fold, P < 0.01 ) and late (2.11 ± 0.28 fold, P < 0.01 ) apoptosis of VSMCs as compared with HG. Yixintongmai inhibited the expression of B-cell lymphoma 2 (0.83 ± 0.07 fold, P < 0.01 ) and stimulated the activity of cleaved-capase-3/caspase-3 (2.00 ± 0.12 fold, P < 0.05 ) as compared with HG. Yixintongmai inhibited reactive oxygen species generation (0.46 ± 0.03 fold, P < 0.01 ) and the expression of NADPH oxidase-1 (0.84 ± 0.04 fold, P < 0.001 ), nuclear factor-kappa B (NF-κB) p65 (0.71 ± 0.07 fold, P < 0.001 ), phosphorylated NF-κB p65 (0.39 ± 0.02 fold, P < 0.0001), and inhibited nuclear translocation of NF-κB p65 (0.87 ± 0.03 fold, P < 0.001 ) in VSMCs as compared with HG. Conclusions. Yixintongmai inhibits the proliferation and migration and promotes the apoptosis of VSMCs cultured with HG, which suggests the potential anti-atherosclerotic effects of this traditional Chinese medicine.

Targeting Reactive Oxygen Species in Atherosclerosis via Chinese Herbal Medicines

“…By measuring cell viability in vitro, Shen et al examined the Buyang Huanwu decoction (BYHWD) played a vital role in protecting neurons against cerebral ischemic injury by downregulation of the expression of NOX-4 and reducing the production of ROS [ 94 ]. Guo et al [ 95 ] found that Yixingtongmai (YXTM) could inhibit the expression of matrix metalloproteinase 9 (MMP-9), nuclear factor kappa-B (NF- κ B), and B cell lymphoma-2 (Bcl-2) and stimulate the activity of caspase 3, suggesting its effects in preventing the development and progression of atherosclerosis. Wang et al [ 96 ] demonstrated that the herbal compound Tongxinluo (TXL) significantly increased myocardial capillary density and alleviated oxidative stress injury in heart failure by inhibiting the expression of NOX-4 and upregulating cardiac nitrite content and the protein expression of vascular endothelial growth factor (VEGF), p-VEGFR2, p-phosphoinositide 3-kinase (p-PI3K), p-AKT, (p-eNOS), andh oxygenase-1 (HO-1).…”