Figure 5.

Berberine induces T cell apoptosis via the mitochondrial apoptosis pathway. (A) (i) KEGG functional categories of differentially expressed genes following berberine and saline treatment. The Y-axis represents the KEGG functional categories. (ii) KEGG analysis of the significantly altered signaling pathways in cell growth- and death-associated genes. The X-axis represents the rich ratio of the number of differentially expressed genes and the Y-axis represents the KEGG pathways. (iii) qPCR analysis of Bcl-2 and TNF-α mRNA expression in SPCs collected from heart transplant recipients treated with berberine or not. (B) T cell apoptosis assays in vivo. (i) SPCs and (ii) LNCs were collected at POD 7. The percentages of apoptotic CD4⁺ and CD8⁺ T cells were determined by flow cytometry (n = 3 mice/group). (C) T cell apoptosis assay in vitro. T cells from naïve C57BL/6 mice were co-stimulated with anti-CD3/CD28 Abs in the absence or presence of berberine. The percentages of apoptotic (i) CD4⁺ and (ii) CD8⁺ T cells were determined by flow cytometry. (D) Berberine activates the mitochondrial apoptosis pathway in vivo. (i) Relative protein expression of Bcl-2, Bax, cytochrome c, cleaved-caspase-3, and cleaved-PARP in SPC. (ii) β-actin was used as a loading control (n = 3 mice/group), and OD values (relative to β-actin) are presented as means ± SEMs. *p < 0.05, **p < 0.01, ***p < 0.001 compared to the normal saline-treated group. (E) Berberine activates the mitochondrial apoptosis pathway in vitro. Relative protein expression of Bcl-2, Bax, cytochrome c, cleaved-caspase-3, and cleaved-PARP expression in CD3⁺ T cells. (ii) β-actin was used as a loading control; OD values (relative to β-actin) are presented as means ± SEMs. SPCs, spleen cells; LNCs, lymph node cells; POD, post-operative day. *p < 0.05, **p < 0.01, ***p < 0.001 compared to the PC group.