Angiotensin-(1-7) ameliorates high glucose-induced vascular endothelial injury through suppressing chloride channel 3

Figures & data

Figure 1. Ang- [1–7] alleviates HG-induced injuries of bEnd3 cells through regulating CIC-3 expression via Mas receptor. (a) Growth rate of bEnd3 cells in different groups. (b) Immunostaining analysis of CIC3 protein in bEnd3 cells. Green indicated CIC3 protein while blue indicated nucleus. (c) WB analysis of CIC3 protein level in bEnd3 cells treated with or without Ang- [1–7]. (d) WB analysis of CIC3 protein level in bEnd3 cells treated with or without siRNAs. * indicates P < 0.05 compared to NG group while # indicates P < 0.05 compared to HG group.

Figure 2. Ang- [1–7] decreases the concentration of chloride in bEnd3 cells under HG condition. (a) Chloride level in bEnd3 cells detected by MQAE probe using a flow cytometer. (b) Quantification of chloride level in bEnd3 cells detected by MQAE probe. * indicates P < 0.05 compared to NG group while # indicates P < 0.05 compared to HG group.

Figure 3. Ang- [1–7] inhibits HG-induced bEnd3 cell apoptosis through suppressing CIC-3 via Mas. (a) Growth rate of bEnd3 cells in different groups. (b) WB analysis of cleaved caspase3 and cytochrome C protein levels in bEnd3 cells. (c) Flow cytometric analysis of apoptosis in bEnd3 cells. * indicates P < 0.05 compared to NG group while # indicates P < 0.05 compared to HG group.

Figure 4. Ang- [1–7] ameliorates HG-reduced mitochondrial potential through downregulating CIC-3 via Mas in bEnd3 cells. (a) JC-1 dye-stained bEnd3 cells were analyzed for the mitochondrial potential. (b) Quantification of fluorescence for JC-1 dye detected by a flow cytometer. * indicates P < 0.05 compared to NG group while # indicates P < 0.05 compared to HG group.

Figure 5. Ang- [1–7] ameliorates HG-induced endothelial injury through downregulating CIC-3 via Mas in bEnd3 cells. (a) ROS level in bEnd3 cells detected by DCFH-DA probe using a flow cytometer. (b) Quantification of ROS level in bEnd3 cells detected by DCFH-DA probe. (c) Quantification of NO level in bEnd3 cells. (d) Quantification of GSH level in bEnd3 cells. * indicates P < 0.05 compared to NG group while # indicates P < 0.05 compared to HG group.

Figure 6. Ang- [1–7] preserves HG-induced endothelial injury through suppressing cytokine production via Mas-CIC3 axis in bEnd3 cells. (a) WB analysis of eNOS, NF-κb, TNFα protein levels in bEnd3 cells. (b) Levels of released IL-1β, IL-6 and IL-8 in bEnd3 cells. * indicates P < 0.05 compared to NG group while # indicates P < 0.05 compared to HG group.