Ginkgolide B Mediated Alleviation of Inflammatory Cascades and Altered Lipid Metabolism in HUVECs via Targeting PCSK-9 Expression and Functionality
Figure 1
(a) Ox-LDL alters the expression of SREBP-2, PCSK-9, and LDL-R in HUVECs. The HUVECs were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM) containing 10% heat-inactivated FBS, 2 mM glutamine, and antibiotics at 37°C in a humidified 5% CO2 atmosphere. Thus, cultured cells were either treated with native LDL or Ox-LDL at different concentrations 25, 50, 75, and 100μg/ml designated as Ox-LDL 25, Ox-LDL 50, Ox-LDL 75, and Ox-LDL 100, respectively, for gene expression analysis. Significantly different from native LDL at <0.001. Nonsignificantly different from native LDL at >0.05. (b) Ginkgolide B amends the Ox-LDL-induced altered expression of SREBP-2, PCSK-9, and LDL-R in HUVECs. The HUVECs were pretreated with 20-100 μg/ml of Ginkgolide B (designated as GB 20, GB 40, GB 80, and GB 100) for 24 h and then all the groups except control were exposed to Ox-LDL (75μg/ml) for another 24 h followed by m-RNA extraction for qRT-PCR analysis. Significantly different from control HUVECs at <0.001. Significantly different from control HUVECs at <0.01. Significantly different from control HUVECs at <0.05. Nonsignificantly different from control HUVECs at >0.05. Significantly different from Ox-LDL at <0.001. (c) Ox-LDL upregulates the expression of LOX-1 and NOX-4 in HUVECs. The HUVECs were cultured and treated with Ox-LDL as described in (a). Significantly different from native LDL at <0.001. (d) Ginkgolide B downregulates the Ox-LDL-induced expression of LOX-1 and NOX-4 in HUVECs. The HUVECs were cultured and treated with Ginkgolide B and Ox-LDL as described in (b). Significantly different from control HUVECs at <0.001. Significantly different from control HUVECs at <0.01. Significantly different from control HUVECs at <0.05. Nonsignificantly different from control HUVECs at >0.05. Significantly different from Ox-LDL at <0.001. All the values in (a), (b), (c), and (d) have been expressed as mean ± SEM of three independent experiments.