Research Paper Volume 13, Issue 5 pp 7096—7119

Berberine-induced TFEB deacetylation by SIRT1 promotes autophagy in peritoneal macrophages

TFEB nuclear translocation and autophagy induced by berberine inhibit apoptosis. (A) CCK-8 assay was used to detect cell viability after different treatments (100 μmol/L berberine, 10 h incubation). Data was analyzed by one-way ANOVA with Tukey HSD post-hoc test (vs. Control group). Analysis of variance and Student-Newman-Keuls post hoc tests were used to compare two group (vs. berberine group). (B) The expression of apoptosis-related proteins BAX, Bcl-2, and cytochrome C was detected by western blotting following different treatments (100 μmol/L berberine, 10 h incubation). Data was analyzed by one-way ANOVA with Tukey HSD post-hoc test (vs. Control group). Analysis of variance and Student-Newman-Keuls post hoc tests were used to compare two group (vs. berberine group). (C) Changes in the number of apoptotic peritoneal macrophages were detected by flow cytometry (100 μmol/L berberine, 10 h incubation). Data was analyzed by one-way ANOVA with Tukey HSD post-hoc test (vs. Control group). Analysis of variance and Student-Newman-Keuls post hoc tests were used to compare two group (vs. berberine group). (D) The mRNA expression of apoptosis-related genes BAX, Bcl-2, and cytochrome C was detected by qRT-PCR after different treatments (100 μmol/L berberine, 10 h incubation). Data was analyzed by one-way ANOVA with Tukey HSD post-hoc test (vs. Control group). Analysis of variance and Student-Newman-Keuls post hoc tests were used to compare two group (vs. berberine group). (E) Changes in the level of released cytochrome C in peritoneal macrophages were detected using MitoTracker Green and anti-cytochrome C immunofluorescence staining. Scale bar, 20 μm (100 μmol/L berberine, 10 h incubation). All values are expressed as mean ± SD (error bars) of three independent experiments. n = 3; *p **p ***p #p

Figure 5. TFEB nuclear translocation and autophagy induced by berberine inhibit apoptosis. (A) CCK-8 assay was used to detect cell viability after different treatments (100 μmol/L berberine, 10 h incubation). Data was analyzed by one-way ANOVA with Tukey HSD post-hoc test (vs. Control group). Analysis of variance and Student-Newman-Keuls post hoc tests were used to compare two group (vs. berberine group). (B) The expression of apoptosis-related proteins BAX, Bcl-2, and cytochrome C was detected by western blotting following different treatments (100 μmol/L berberine, 10 h incubation). Data was analyzed by one-way ANOVA with Tukey HSD post-hoc test (vs. Control group). Analysis of variance and Student-Newman-Keuls post hoc tests were used to compare two group (vs. berberine group). (C) Changes in the number of apoptotic peritoneal macrophages were detected by flow cytometry (100 μmol/L berberine, 10 h incubation). Data was analyzed by one-way ANOVA with Tukey HSD post-hoc test (vs. Control group). Analysis of variance and Student-Newman-Keuls post hoc tests were used to compare two group (vs. berberine group). (D) The mRNA expression of apoptosis-related genes BAX, Bcl-2, and cytochrome C was detected by qRT-PCR after different treatments (100 μmol/L berberine, 10 h incubation). Data was analyzed by one-way ANOVA with Tukey HSD post-hoc test (vs. Control group). Analysis of variance and Student-Newman-Keuls post hoc tests were used to compare two group (vs. berberine group). (E) Changes in the level of released cytochrome C in peritoneal macrophages were detected using MitoTracker Green and anti-cytochrome C immunofluorescence staining. Scale bar, 20 μm (100 μmol/L berberine, 10 h incubation). All values are expressed as mean ± SD (error bars) of three independent experiments. n = 3; *p < 0.05, **p < 0.01, and ***p < 0.001 versus control. #p < 0.05 versus berberine group.