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. 2022 Aug 15;14(16):3338. doi: 10.3390/nu14163338

Figure 2.

Figure 2

TMAO increased the ROS level and inflammatory factors by inhibiting SIRT1 expression. (A–D).The protein level (upper panels) and transcript level (lower panels) of SIRT1 were determined in VSMCs (A,C) and HUVECs (B,D) treated with 1000 μM TMAO (A,B) and transfected with pCMV-Tag 2B- SIRT1 plasmid (C,D) for 24 h by western blot and quantitative real-time PCR. (E,F) VSMCs (E) and HUVECs (F) were treated with 1000 μM TMAO, SIRT1 vector and 1000 μM TMAO plus SIRT1 vector. The total ROS levels were determined using fluorescence intensity of DCFH-DA staining by flow cytometry. (G,H) The protein level (upper panels) and transcript level (lower panels) of inflammation-related markers IL-1β, TNFα, MMP9 were determined in VSMCs (H) and HUVECs (F) treated with 1000 μM TMAO, SIRT1 vector and 1000 μM TMAO plus SIRT1 vector for 24 h. Values are the mean ± SD; n = 3; two-way ANOVA followed by a Tukey post hoc test. * p < 0.05; ** p < 0.01 vs. Control; # p < 0.05; ## p < 0.01 vs. TMAO group; + p < 0.05; ++ p < 0.01 vs. SIRT1 group.