The Protective Effect Of SQS In High Glucose-induced Injury Of Endothelial Cell Mediated By Anion Exchanger-2

Objective:To investigate the protection of sasanquasaponin(SQS) on injury of human umbilical vein cells (HUVECs) induced by high glucose and its possible mechanisms. And provide theoretical and experimental basis to further study the mechanism of DM.Methods:<1> The dose-effect of SQS on injury of HUVECs induced by high glucose:HUVECs (CRL-1730) were randomly divided into eight groups:①Control group,②HG group,③0.1μM SQS group(SQS-0.1),④0.5μM SQS group (SQS-0.5),⑤1μM SQS group(SQS-1),⑥5μM SQS group(SQS-5),⑦10μM SQS group(SQS-10),⑧50μM SQS group (SQS-50),establishment of high glucose injury model except control group.and cultured for48hours. Cell viability,LDH level were measured by a colorimetric method.<2>The possible protection mechanism of SQS:HUVECs have been randomly divided into four groups:①Control group,②HG group,③SITS group,④SQS-5group (the best concentration). SOD,GSH-Px and MDA in HUVECs were measured by a colorimetric method. The percentage of apoptosis was measured by flow cytometry. Meanwhile, intracellular ROS and CI-levels were also measured by flow cytometry. Expression of SOD and AE2protein were detected by Western blotting while the expression of AE2mRNA was detected by RT-PCR.Results:1.HUVECs were incubated with high glucose for48h,cell viability significantly decreased while the level of LDH were significantly increased compared with the Control group (P<0.05). Six different concentrations of SQS Group,cell viability was significantly increased while the levels of LDH decreased compared with the HG (P<0.05),there was no difference between SQS-5and SQS-10,so SQS-5was the best dose on injury of HUVECs induced by high glucose.2. SITS group,SQS group were incubated with high glucose for48h, activities of SOD,GSH-Px and the expression of SOD Protein were remarkably increased while MDA,ROS,apoptosis, concentration of CI-were decreased compared with HG (P<0.05).3. The expression of AE22mRNA and and its protein were remarkably increased in HG group(P<0.01,vs control group).SQS group could down-regulated expression of AE2mRNA and and its protein.SITS group was similar to the SQS group.Conclusion:SQS can protect against injury of HUVECs induced by high glucose mediated by AE2,the mechanism may be associated with inhibition of Cl-overload, anti-oxidative stress and anti-apoptosis.