Effects of high glucose and the culture fluid with high glucose condition on vascular endolthelial cells
Liu Chang-qi
Abstract
Liu Chang-qi
Abstract
Objective To observe the effects of high glucose on the expression of HMGB1 in cultured vascular endothelial cells(VECs) and the effects of the culture fluid with high glucose condition on oxidative stress.Methods 1.The human umbilical veins endothelial line(Ea.hy926) were cultured in vitro.VECs were cultured with glucose in different concentrations(5.6 mmol/L,20 mmol/L and 33mmol/L) for 48h.The levels of HMGB1 in the supernatant were measured by Western blot.The cell viability within 48h was examined by trypan blue exclusion test.2.VECs were divided into two groups: high glucose group(33mmol/l) and the mixed group(the supernatant of 33mmol/L group plus 33mmol/L high glucose medium).24 hours later,the levels of MDA and SOD were measured by UV-Spectrometer.Results 1.Compared with normal control,the expression of HMGB1 was significantly increased in the supernatant of high glucose group(P0.05).Within 48h,no differences in cell viability were found compared with control group by Trypan blue exclusion test.2.The level of MDA was significantly higher in the mixed group than that in 33mmo/L high glucose group.And the activity of SOD was lower in the mixed group than that in high glucose group(P0.05).Conclusion High glucose could significantly induce the expression of HMGB1 in cultured VESs.In high glucose condition,the level of HMGB1 increases,which may play a role in oxidative stress and promote the occurrence of cerebrovascular disease.
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Objective To observe the effects of high glucose on the expression of HMGB1 in cultured vascular endothelial cells(VECs) and the effects of the culture fluid with high glucose condition on oxidative stress.Methods 1.The human umbilical veins endothelial line(Ea.hy926) were cultured in vitro.VECs were cultured with glucose in different concentrations(5.6 mmol/L,20 mmol/L and 33mmol/L) for 48h.The levels of HMGB1 in the supernatant were measured by Western blot.The cell viability within 48h was examined by trypan blue exclusion test.2.VECs were divided into two groups: high glucose group(33mmol/l) and the mixed group(the supernatant of 33mmol/L group plus 33mmol/L high glucose medium).24 hours later,the levels of MDA and SOD were measured by UV-Spectrometer.Results 1.Compared with normal control,the expression of HMGB1 was significantly increased in the supernatant of high glucose group(P0.05).Within 48h,no differences in cell viability were found compared with control group by Trypan blue exclusion test.2.The level of MDA was significantly higher in the mixed group than that in 33mmo/L high glucose group.And the activity of SOD was lower in the mixed group than that in high glucose group(P0.05).Conclusion High glucose could significantly induce the expression of HMGB1 in cultured VESs.In high glucose condition,the level of HMGB1 increases,which may play a role in oxidative stress and promote the occurrence of cerebrovascular disease.
Key concepts: Trypan blue, L-Glucose, Oxidative stress, Western blot, HMGB1, Andrology, Viability assay, Cell culture