Figure 6.

Hsp90 induction by aspirin relieved the heat-stressed damage. Data represent the means ± SD for three independent experiments. (A) After treatment with aspirin of different concentrations for 24 h, CCK-8 was used to detect cell viability. (B) CMVECs were administrated with aspirin of 1 mg/mL or aspirin plus sodium bicarbonate (ASA: 1 mg/mL; the mole ratio of ASA and NaHCO₃ = 1:1) for different amounts of time. Hsp27, Hsp70 and Hsp90 levels were analyzed with Western blot analysis. The relative abundance of the tested proteins was normalized to that of β-actin. 1: ASA for 0 h; 2. ASA for 0.5 h; 3. ASA for 1 h; 4. ASA for 2 h; 5. ASA for 4 h; 6. ASA and NaHCO₃ (ASA-Na) for 0 h; 7. ASA-Na for 0.5 h; 8. ASA-Na for 1 h; 9. ASA-Na for 2 h; and 10. ASA-Na for 4 h; (C) CMVECs were administrated with aspirin of 1 mg/mL for different time. The specific kits were used to detect cellular LPO and MDA levels and NO release into the supernatant. (D–G) CMVECs with or without aspirin treatment for 2 h were treated with heat stress for 5 h and harvested for corresponding detection. (D) Hsp27, Hsp70 and Hsp90 levels were analyzed with Western blot analysis. The relative abundance of the tested proteins was normalized to that of β-actin. (E) Flow cytometry was used to detect the cellular apoptosis rate. (F) H. E. staining was conducted and observed under the light microscope. a–d shows the representative pictures from Con, ASA, HS, and ASA+HS, respectively. Bar = 20 µm. Asterisk point to necrosis, and arrowhead mark degeneration. (G) The specific kits were used to detect cellular LPO and MDA levels and NO release into the supernatant. Con, cells without aspirin treatment and heat stress; ASA, cells treated with aspirin; HS, heat-stressed cells without aspirin; ASA + HS, heat-stressed cells following aspirin treatment. The differences of the data of cells with different treatment vs. that of the Con are indicated by * p < 0.05 and ** p < 0.01. The differences of the data of cells treated by ASA + HS vs. that in ASA or HS are indicated by # p < 0.05 and ## p < 0.01. The difference of the data of cells treated by ASA vs. that in HS are indicated by & p < 0.05 and && p < 0.01.