Fig 1: Different from TSA, Mocetinostat, a class-? selective HDAC inhibitor did not attenuate UA-stimulated endothelial dysfunction in HUVECs. A, HDAC1,2,3,6 activity detected by HDAC kits (n = 5). B, TNF-a, IL-1ß, and IL-6 mRNA level detected by RT-qPCR (n = 5). C, NO production (n = 5). D, RT-qPCR analysis of FGF21 (n = 5). E, WB analysis of FGF21 (n = 4). F, Level of histone H3 acetylation on the FGF21 promoter region, as analyzed by ChIP (n = 4). One-way ANOVA followed by Tukey's post hoc multiple-group comparisons. *P < 0.05, **P < 0.01, ***P < 0.001 versus CON; #P < 0.05, ##P < 0.01, ###P < 0.001 versus 12 mg/dL UA; $P < 0.05, $$P < 0.01, $$$P < 0.001 versus 12 mg/dL UA+TSA high.
Fig 2: TSA or TubA alleviated renal inflammation and dysfunction in hyperuricemic mice. A, Serum BUN (n = 7–8). B, Serum CRE (n = 8). C, RT-qPCR analysis of TNF-a and IL-6 in renal tissues (n = 5). D, Representative images of mice renal tissue stained by H&E, Masson, and PAS (magnification × 200), and percentage of fibrotic area was analyzed. E, WB analysis HDAC1, 2, 3, and 6 (n = 5). F, WB analysis of p-AKT/AKT, p-eNOS/eNOS, and p-FoxO3a/FoxO3a (n = 5). G, WB analysis of FGF21, CD31, and a-SMA (n = 5). One-way ANOVA followed by Tukey's post hoc multiple-group comparisons. *P < 0.05, **P < 0.001, ***P < 0.001 versus CON; #P < 0.05, ##P < 0.01, ###P < 0.001 versus model; $P < 0.05, $$P < 0.01 versus TSA.
Fig 3: TSA or TubA alleviated aortic inflammation and dysfunction in hyperuricemic mice. A, Blood pressure between the groups (n = 15). B, Serum UA (n = 7–8). C, Serum NO (n = 8). D, Serum FGF21 (n = 7–8). E, Serum TNF-a and IL-6 (n = 7–8). F, Levels of TNF-a and IL-6 mRNA in aorta tissue were determined by RT-qPCR (n = 5). G, Vasorelaxant responses to Ach and SNP in mice aortic rings precontracted with phenylephrine (n = 5). H, Representative images of mice aorta histopathology through H&E staining (magnification × 200) and used for analysis of media thickness. I, WB analysis of HDAC1, 2, 3, and 6 in aorta tissue (n = 5). J, WB analysis of p-AKT/AKT, p-eNOS/eNOS, and p-FoxO3a/FoxO3a in aorta tissue (n = 5). K, WB analysis of FGF21, CD31, and a-SMA in aorta tissue (n = 5). One-way ANOVA followed by Tukey's post hoc multiple-group comparisons. *P < 0.05, **P < 0.01, ***P < 0.001 versus CON; #P < 0.05, ##P < 0.01, ###P < 0.001 versus model; $P < 0.05, $$P < 0.01 versus TSA.
Fig 4: FGF21 knockdown abrogated the effects of TSA on UA-induced inflammation, oxidative stress, endothelial dysfunction, and endothelial–interstitial transformation in HUVECs. A, RT-qPCR analysis of FGF21 (n = 3). B, RT-qPCR analysis of TNF-a, IL-1ß, and IL-6 (n = 4). C, ROS generation (n = 3). D, NO production (n = 5). E, Levels of p-AKT/AKT and p-eNOS/eNOS measured by WB (n = 4). F, RT-qPCR analysis of CD31 and a-SMA (n = 4). G, WB analysis of CD31 and a-SMA (n = 4). One-way ANOVA followed by Tukey's post hoc multiple-group comparisons. *P < 0.05, **P < 0.01, ***P < 0.001 versus CON; ###P < 0.001 versus 12 mg/dL UA; $$P < 0.01, $$$P < 0.001 versus 12 mg/dL UA+TSA high; &P < 0.05, &&P < 0.01, &&&P < 0.001 versus 12 mg/dL UA+TSA high+NC.
Fig 5: High UA levels stimulated HDAC6 and inhibited FGF21 expression in HUVECs. A, NO production detected by NO assay kit (n = 5). B, HDAC6 mRNA expression determined by RT-qPCR (n = 5). C, HDAC6 protein expression determined by WB analysis (n = 4). D, FGF21 mRNA level measured by RT-qPCR (n = 5). E, FGF21 protein level measured by WB analysis (n = 4). One-way ANOVA followed by Tukey's post hoc multiple-group comparisons. *P < 0.05, **P < 0.01, ***P < 0.001 versus CON; #P < 0.05, ##P < 0.01, ###P < 0.001 versus 6 mg/dL UA.
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