Fig 1.
Effect of irbesartan on cell viability and on oxidative stress in HL-1 hypoxic cells.
(A) Cardiomyocytes viability was determined by MTT assay in normoxia (CTRL), hypoxia alone (Hypoxia) and in combination with a pre-treatment or post-treatment with 16 h of irbesartan (Irb 10, 50, 100 μM). (B) SOD activity was determined by an ELISA assay. (C) Catalase protein expression was measured by western blot. All data are presented as mean ± S.E.M. of three independent experiments (*P<0.05 versus CTRL; #P<0.05 versus hypoxia, § P<0.05 versus irbesartan 10 μM).
Fig 2.
Effect of irbesartan pre-treatment on oxidative stress, on cardiac marker BNP and on TLRs mRNA expression.
(A) The release of peroxynitrite was measured by Griess assay. (B) mRNA levels BNP was evaluated by RT-qPCR. (C) Western blot analysis of BNP protein, blot is representative from three independent experiments. (D) TLR2 and (E) TLR4 mRNA expressions were measured by RT-qPCR. TLR2 an TLR4 mRNA levels were normalized relative to GAPDH mRNA levels. Data represent the mean ± S.E.M. of three independent experiments (*P<0.05 versus CTRL; #P<0.05 versus hypoxia, § P<0.05 versus irbesartan 10 μM).
Fig 3.
Role of irbesartan pre-treatment on inflammatory mediators and on cytokines activity.
(A) iNOS TNF-alpha and IL-17 proteins expressions were measured by western blot. GAPDH was used as internal standard. Representative blot from three independent experiments. Data are expressed as relative densitometric units and represent the mean ± S.E.M. of three independent experiments (*P<0.05 versus CTRL; #P<0.05 versus hypoxia, § P<0.05 versus irbesartan 10 μM). (B) TNF-alpha and (C) IL-17 activity was measured by ELISA. Final concentration of each cytokine was expressed as pg/mg and represent the mean ± S.E.M. of three independent experiments (*P<0.05 versus CTRL; #P<0.05 versus hypoxia).