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

Figure 3

Yixintongmai promotes the apoptosis and inhibits the expression of Bcl-2 and the activity of caspase-3 of VSMCs cultured with high glucose. NG: normal glucose (5.5 mM); HG: high glucose (25 mM); HG + Y: high glucose (25 mM) and yixintongmai powder (360 μg/ml); MG: normal glucose (5.5 mM) and mannitol (19.5 mM); Normal VSMCs stained with JC-1 emitted orange-red fluorescence with a little green fluorescence, reflecting the condition of mitochondrial membrane potentials. The increased ratio of orange-red fluorescence and green fluorescence indicates the elevation of mitochondrial membrane potentials, due to the increase of aggregated JC-1 within mitochondria. Fluorescence intensity in VSMCs was quantified using Image Pro Plus/IOD. The mitochondrial membrane potentials shown by the fluorescence intensity of VSMCs under different conditions (a); densitometry quantification of fluorescence intensity (c); flow cytometry Annexin V-FITC-PI assay in VSMCS in different groups (b, d, e). Lower right quadrant represents cells that undergo early apoptosis. Upper right quadrant represents cells that undergo late apoptosis. Western blot of Bcl-2 expression (f) and densitometry quantification (g); densitometry quantification values are shown by fold over NG or HG for each time point. Western bolt of caspase-3 and cleaved-caspase-3 expression (f), and densitometry quantification (h); the ratio of cleaved-caspase-3 and caspase-3 protein expression represented the activity of caspase-3. Values are expressed as means ± SD of triplicate experiments. ; : ; NS: not significant (); CCCP: carbonyl cyano-to-chlorobenzene hydrazone (positive control).

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