The Study Of Sodium Deoxycholate In Fat Dissolution And Apoptosis Of Microvascular Endothelial Cells

Objective: To verify the effect of sodium deoxycholate on fat dissolution and investigate sodium deoxycholate's effect on the apoptosis of microvascular endothelial cells so that we can supply theoretical basis for reducing the damage of sodium deoxycholate to blood vessels and increase security of injectable sodium deoxycholate.Methods: 1.Cell Counting Kit(CCK-8 assay)and LDH assay were applied to investigate the damage of sodium deoxycholate to adipocytes.2.Tests including CCK-8 assay?Western Blot?Real-time PCR?Annexin V/PI staining were utilized to confirm the injury action of sodium deoxycholate on human dermal microvascular endothelial cells and research the influence of apoptosis involving Caspase-3 activation in this damage process.3.Study the effect of apoptosis during adipocytes damage caused by sodium deoxycholate through Hoechst staining ? Western Blot and CCK-8 assay.4.Observe if the degree of injury and apoptosis of human dermal microvascular endothelial cells change after using sodium deoxycholate associated with Z-DEVD-FMK.Tests include CCK-8 assay?Western Blot?Annexin V/PI staining.Results:1.Different concentrations of sodium deoxycholate could cause the decrease of fat cell viability,the damage of fat cell membrane and at the end induce the number of adipocytes.2.Various concentrations of sodium deoxycholate could cause the decrease of human dermal microvascular endothelial cells.After the microvascular endothelial cells exposing to sodium deoxycholate(0.15mg/m L)for 4?8?12h,Western Blot implied that the expression of Bax as well as Caspase-3 and PARP activation increased.Except 4h,Caspase-9 activation in experiment groups was more than that in control group,while Bcl-2 expression and Caspase-8 activation did not change obviously.When sodium deoxycholate was associated with Z-DEVD-FMK,both Caspase-3 and PARP activation decreased.The result of Real-time PCR showed that when human dermal microvascular endothelial cells exposed to sodium deoxycholate,Bax mRNA increased,while Bcl-2 mRNA did not.Annexin V/PI staining showed that low concentrations of sodium deoxycholate could lead the early apoptosis of human dermal microvascular endothelial cells and high concentrations lead late apoptosis or necrosis increasing.3.Hoechst staining showed sodium deoxycholate could not induce the apoptosis of fat cells.Caspase-3 did not obviously activate.Adding Z-DEVD-FMK to sodium deoxycholate could not make fat cells viability increase.4.Sodium deoxycholate can reduce the damage of sodium deoxycholate to adipocytes and reduce the activation of Caspase-3 as well as PARP and early apoptosis.Conclusion: Sodium deoxycholate can injure human dermal microvascular endothelial cells by apoptosis pathway including Caspase-3 activation,but its physical destructionis still severe.