Fig. 7.
S1P1R activation attenuates acetaldehyde-induced degranulation and renin release in HMC-1 and WT-BMMC but not in ALDH2*2-BMMC. Incubation with SEW2871 (0.1 and 1 µM, 10 minutes) diminishes degranulation and renin release evoked by acetaldehyde (100–300 µM, 20 minutes) in (A) HMC-1 and (B) WT-BMMC but not in (C) ALDH2*2 cells. Pretreatment with W146 (0.1 and 1 µM, 20 minutes), εV1–2 (1 μM, 20 minutes), or GTN (2 μM, 30 minutes) prevents the effect of the S1P1R agonist in (A) HMC-1, (B) WT-BMMC, and (C) ALDH2*2-BMMC cells. Bars are mean ± S.E.M. of the percentage of increase above control (β-HEX and renin in HMC-1, WT-BMMC, ALDH2*2-BMMC, respectively: acetaldehyde, n = 20, 22, 19 and n = 9, 13, 8; SEW2871, n = 9, 10, 7 and 9, 10, 7; SEW2871 + W146, n = 17, 14, 13 and n = 7, 10, 6; SEW2871 + εV1–2, n = 5, 5, 5 and n = 5, 5, 5; and SEW2871 + GTN, n = 7, 5, 5 and n = 5, 5, 5). β-HEX and renin content was measured in the supernatants at the end of acetaldehyde incubation. Basal β-HEX in HMC-1, WT-BMMC, and ALDH2*2-BMMC was 3.14 ± 0.41 (n = 14), 2.87 ± 0.27 (n = 23), and 2.92 ± 0.31 (n = 21), the percentage of total, respectively; basal renin release (Ang-I formed) for HMC-1 was 18.26 ± 4.12 (n = 17) ng/h/mg protein, and for WT-BMMC and ALDH2*2-BMMC was 0.25 ± 0.04 (n = 29) and 0.49 ± 0.15 (n = 22) µg/h/mg protein, respectively. *P < 0.05; **P < 0.01; ***P < 0.001 by unpaired t test.