Shuxuetong Inhibits Bim-dependent Apoptosis of Cerebellar Granule Neurons / 中山大学学报(医学科学版)

ABSTRACT

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.