Figure 1.
The protective effect of TLQP-21 to HUVECs with cytoxic injury.
(A) Annexin-V-PI staining to assess the apoptotic ratios of HUVECs with TLQP-21 treated alone or under high glucose (HG) concentration (30 mmol/L) incubation for 72 h with or without TLQP-21(10–100 nM). (B) Cell viability of HUVECs under HG concentration (30 mmol/L) incubation for 72 h with or without TLQP-21(10–100 nM) protection.* p<.05,** p<.01 vs. saline treated group. (C) Cell viability of HUVECs with etoposide (0.2 mM) treatment for 72 h with or without TLQP-21(10–100 nM) protection. * p<.05, ** p<.01 vs. saline treated group. All data were collected from three independent experiments and are presented as the mean ±SD.
Figure 2.
TLQP-21 increases the NADPH level without promoting cell growth.
(A) The absorbance value of MTS assay with different treatments; the mannitol group was set as hyperosmosis control. *p<.05, **p<.01 vs. NC group; # p<.05, ##p<.01 vs. HG group. (B) Cell proliferation curve of HUVECs with different treatments. #p<.05, ##p<.01 NC&TLQP-21 group vs. HG group; *p<.05, **p<.01 HG+TLQP-21 group vs. HG group. (C) NADPH level of HUVECs with different treatments. **p<.01 vs. NC group; ##p<.01 vs. TLQP-21(10 nM) group. (D) Comparison of the NADPH level of HUVECs with and without TLQP-21(50 nM) treatment; the mannitol group was set as a hyperosmosis control. *p<.05, *p<.01 vs. TLQP-21 withdraw group. All data were collected from three independent experiments and are presented as the mean ±SD.
Figure 3.
TLQP-21 treatment (72 h) protects the HUVECs from oxidative stress by intercellularly rebalancing the redox state.
(A) GSH level of HUVECs with different treatments. (B) SOD activity of HUVECs with different treatments. (C) Catalase activity of HUVECs with different treatments. (D) DCF intensity presents the ROS levels of HUVECs with different treatments. (E) TBARs level of HUVECs with different treatments. *p<.05, **p<.01 vs. HG group; ##p<.01 vs. NC group. All data were collected from three independent experiments and are presented as the mean ±SD.
Figure 4.
TLQP-21 increases NADPH expression, but depends on G6PD.
(A) G6PD activity of HUVECs with different treatments. **p<.01 vs. NC group. (B) mRNA level of G6PD with different treatments. **p<.01 vs. NC group (C) Western-blot analysis shows that G6PD expression decreased with G6PD siRNA. (D) G6PD activity decreases with G6PD siRNA. **p<.01 vs. scramble siRNA group. (E) G6PD siRNA attenuates the upregulation of NADPH when treated with TLQP-21. **p<.01 vs. scramble siRNA group. All data were collected from three independent experiments and are presented as the mean ±SD.