الملخص
ObjectiveTo investigate the effect and mechanism of Shuxuetong and its main component hirudin on the apoptosis of cerebellar granule neurons (CGNs) in Sprague-Dawley(SD) rats. MethodsCGNs incubated in vitro for 7 days were divided into survival control group or 25 K group (cultured in medium containing 25 mmol/L KCL) and apoptosis group or 5 K group (cultured in medium containing 5 mmol/L KCL). CGNs were separately treated with proportionally diluted and different concentrations of Shuxuetong (1/50, 1/40, 1/30, 1/20 and 1/10) and the corresponding different concentrations of hirudin (2, 2.5, 3.34, 5 and 10 U / mL). Hoechst staining was performed to analyze the apoptosis. Western blot was used to detect the expression levels of Cleaved Caspase-3, Bim and VEGF. ResultsHoechst staining showed that 5 K group had a higher apoptosis rate than 25 K group. In 25 K group, there was no significant change in the apoptosis rate between neurons treated with different concentrations of Shuxuetong and hirudin, but significant changes was found in 5 K group and the higher the concentration, the lower the apoptosis rate. Western blot results revealed that, compared with control neurons in 5 K group, Shuxuetong injection and hirudin treatments resulted in a decrease of Cleaved Caspase-3 and Bim expression, but an increase of VEGF protein. ConclusionsShuxuetong and its main component hirudin inhibits the apoptosis of CGNs through suppressing proapoptotic BH3-only protein Bim.