Effect of tanshinone IIA on inflammatory injury induced by trimethylamine oxide in human umbilical vein endothelial cells

Abstract

Objective To investigate the inhibitory effect of tanshinone ⅡA on trimethylamine oxide(TMAO)induced inflammatory injury of human umbilical vein endothelial cells(HUVECs), and its effect on atherosclerosis induced by trimethylamine oxide. Methods HUVECs were cultured in vitro, the experiment was divided into normal group, TMAO, TMAO+ tanshinone Ⅱ A 10 μg/ml group and TMAO+ tanshinone Ⅱ A 20 μg/ml group.The levels of NO, lactate dehydrogenase(LDH)in the cell culture supernatant of each group were detected.The expression levels of cell closure protein, eNOS and ICAM-1 protein were detected.The levels of interleukin-1β(IL-1β), interleukin-6(IL-6)and C-reactive protein(CRP)in the cell culture supernatant were further detected. Results The released NO in the TMAO group[(411.40±50.10)μmol/L]was significantly lower than[(755.40±45.60)μmol/L]in the normal group, and the released LDH[(10.50±1.53)U/L]was higher than[(4.20±0.34)U/L]in the normal group.The quantity of released NO in TMAO+ tanshinone Ⅱ A 10 μg/ml group[(519.20±45.30)μmol/L]and in TMAO+ tanshinone Ⅱ A 20 μg/ml group[(557.80±50.80)μmol/L]were both higher than[(411.40±50.10)μmol/L]in TMAO group.The released LDH in TMAO+ tanshinone Ⅱ A 10 μg/ml group[(8.20±1.05)U/L]and TMAO+ tanshinone Ⅱ A 20 μg/ml group[(7.40±0.67)U/L]were both lower than[(10.50±1.53)U/L]in TMAO group, and the difference was statistically significant(P<0.05). In the TMAO group, the expression of cell closure protein and eNOS protein was reduced, while the expression of ICAM-1 protein was increased.TMAO also significantly increased the levels of IL-1β, IL-6 and CRP in the supernatant of endothelial cells.In TMAO+ tanshinone Ⅱ A 10 μg/ml group, after treatment, endothelial cell closure protein and eNOS protein were up-regulated, and ICAM-1 protein was down-regulated.The IL-1β, IL-6 and CRP in TMAO+ tanshinone Ⅱ A 10 μg/ml group[(0.24±0.23)ng/ml, (4.54±0.41)ng/ml and(2.03±0.18)ng/ml]and in TMAO+ tanshinone Ⅱ A 20 μg/ml group[(0.22±0.25)ng/ml, (4.33±0.29)ng/ml and(1.95±0.14)ng/ml]were both lower than[(3.01±0.41)ng/ml, (5.10±0.48)ng/ml and(2.48±0.29)ng/ml]in TMAO group, TMAO+ tanshinone Ⅱ A 20 μg/ml group reduced more, and the difference was statistically significant(P<0.05). Conclusion Tanshinone Ⅱ A shows concentration dependence on up-regulation of cell supernatant fluid NO level, reduces the cell supernatant LDH release, increases cell closure protein and eNOS protein expression, reduces cells ICAM-1 expression, and reduces the cell supernatant in IL-1β, IL-6, CRP levels.Tanshinone Ⅱ A can inhibit inflammation TMAO induced endothelial cell damage, which is worth of clinical promotion. Key words: Tanshinone ⅡA; Trimethylamine oxide; Human umbilical vein endothelial cells; Inflammatory injury