Effects Of Ginkgo Biloba Extract And Ginkgolides B On Expression Of β-amyloid In Endothelial Cells Incubated With Diabetic Serum

ObjectiveTo investigate the mechanism of the protective effects of endothelial cell which was pretreatmented by Ginkgo biloba extract (GBE), ginkgolides B (GB) and atovastatin (AT) induced by diabetic serum (DS) and to search for the possible mechanism.MethodsTo examine damage of ECV-304 cells which were pretreatmented by GBE and GB induced by DS. Cultured ECV-304 cells were pretreated with GBE (100μg/ml) and GB (10μg/ml) for 24hours, and then exposured to 10% DS for 30minutes, 3hours, 3days. AT acted as positive control. The morphology of ECV-304 cells was observed by microscope. The proliferative activity of ECV-304 was measured by MTT assay. The cells injury was measured by LDH, MDA and NO assays. To detectβ-amyloid (Aβ) by ELISA kits in ECV-304 cells culture medium which were pretreatmented by GBE, GB and AT induced by DS.Results1. To examine damage of ECV-304 cells which were pretreatmented by GBE, GB and AT induced by DS. At 30 minutes after intervention with AT, GB and GBE, all results were not obviously different from DS groups (P>0.05).The proliferative activity of ECV-304, NS group (104.1±2.1, P<0.05) compared with DS group (96.7±2.1) were advanced. GBE group (119.5±1.6, P<0.001), GB group (108.8±1.9, P<0.05) and AT group (120.4±3.7,P<0.001) compared with DS group was advanced at the third day.LDH concentration (U/L), GBE groups (770.6±40.5, P<0.01, 912.2±34.2, P< 0.05), GB groups (861.1±36.5, P<0.05, 955±29.8, P<0.05) compared with DS groups (1054±38.9, 1211±54.32) were degraded at the third hour and the third day, AT groups (896.1±43.6, 1060.9±38.3) were not obviously different from DS groups (P>0.05)。After interfered for 3hours and 3days, the level of MDA (μM) in GBE groups (7.76±0.42, P<0.05; 6.47±0.45, P<0.01), GB groups(8.90±0.17, P<0.05; 8.24±0.37, P<0.01) and AT groups (7.95±0.37, P<0.05; 7.95±0.42, P<0.01) were lower than that in DS groups (9.52±0.13, 12.43±0.62).GBE (93.8±11, P<0.01), GB (144.2±19, P<0.01) and AT (147.2±21, P<0.01) enhanced produce of NO (μM) compare with DS group (50.2±1.9) after intervention for 3 hours. GBE (65.9±0.7, P<0.05) and AT (71.3±2.8, P<0.05) enhanced produce of NO (μM)compared with DS group (41.25±6.4) after intervention for 3 days.2. To detect the concentration of Aβin ECV-304 cells culture medium which were pretreatmented by GBE, GB and AT induced by DS.At three time points, the concentration of Aβ42 (pM) in DS groups (7.43±0.25, P<0.001; 7.51±0.20, P<0.001; 11.64±0.19, P<0.001) compared with the diabetic serum (0.52±0.05) was advanced, the concentration of Aβ40 (pM) in DS groups (9.66±0.21, P<0.001; 11.19±0.53, P<0.001; 55.24±0.27, P<0.001) compared with the diabetic serum (0.74±0.05) was advanced.The concentration of Aβ42 (pM) in GBE groups (2.37±0.17, P<0.001; 2.34±0.11, P<0.001; 2.7±0.10, P<0.001), GB groups (2.37±0.15, P<0.001; 4.1±0.17, P<0.001; 5.7±0.19, P<0.001) and AT groups (5.29±0.23, P<0.001; 5.2±0.09, P<0.001; 4.9±0.17, P<0.001) compared with DS groups (7.41±0.25, 7.51±0.11, 11.6±0.11) were degraded respectively.At all time points the concentration of Aβ40 (pM) in GBE groups (0.27±0.06, P<0.001; 3.68±0.22, P<0.001; 36.4±0.22, P<0.001), GB groups (0.18±0.14, P<0.001; 4.94±0.20, P<0.001; 43.1±0.22, P<0.001) and AT groups (2.81±0.12, P<0.001; 9.63±0.21, P<0.001; 38.9±0.14, P<0.001) compared with DS groups (9.74±0.43, 9.58±0.16, 55.2±0.16) were degraded respectively.ConclusionsAβmay participate the injury of endothelial cells induced by diabetic serum. GBE, GB and AT protect endothelial cells against the damage induced by DS. The beneficial effects of GBE, GB and AT may be related to their regulation to the production and metabolism of Aβ. GBE demonstrates the most obvious protective effects, which suggest that GBE have an important clinical role in intervention of endothelial dysfunction.