Both Stat3 and Stat5 are regulated by mTOR; mTOR signaling can activate Stat3 and inhibit Stat5 [32]

Both Stat3 and Stat5 are regulated by mTOR; mTOR signaling can activate Stat3 and inhibit Stat5 [32]. Phloretin can broadly activate the AMPK pathway in most cells, such as murine preadipocytes [33], murine lung fibroblasts [34], murine osteocytes [35], mouse marrow stromal cells [36], and human umbilical vein endothelial cells (HUVECs) [37]. versus the control group; ## 0.01 and #### 0.0001 versus the 25? 0.01 versus the 50?= 4). (c) Cell cycle of na?ve and anti-CD3/CD28-activated CD4+ T cells was measured in the presence of phloretin (25, 50, and 100? 0.01 and ???? 0.0001 versus the control group; # 0.05 and #### 0.0001 versus the 25? 0.01 versus the 50?= 4). (aCc) The control group was treated with DMSO, and one-way ANOVA followed by Tukey’s multiple comparison test was used in each statistical analysis. 3.2. Phloretin Influences the Differentiation of Th17 and Treg Cells In Vitro We stimulated purified na?ve CD4+ T cells in vitro with Th17-polarizing conditions or Treg-polarizing conditions with different concentrations of phloretin. After 3-day culture, the frequency of Th17 cells and Treg cells was tested by flow cytometry. As Figure 2(a) showed, the cell count of Th17 cells was significantly reduced when cultured with phloretin. In contrast, the number of Treg cells was significantly increased when exposed to phloretin (Figure 2(b)). These results showed that phloretin could influence the differentiation of Th17 and Treg cells. In addition, 50? 0.0001). In consideration of the proliferation inhibition effect for activated CD4+ T cells and the same effect (no significant difference) between 50? 0.05 and ???? 0.0001 versus the Ginsenoside Rb2 control group; #### 0.0001 versus the 25?= 4, one-way ANOVA followed by Tukey’s multiple comparison test). (b) The frequency of Treg cells generated under Treg polarization conditions in the presence or absence of phloretin (25, 50, and 100? 0.01 and ???? 0.0001 versus the control group; #### 0.0001 versus the 25?= 4, one-way ANOVA followed by Tukey’s multiple comparison test). (c) Expression levels of phospho-Stat3 or phospho-Stat5 were examined under Th17 or Treg polarization conditions in the presence or absence of phloretin (50? 0.01 versus the control group (mean Ginsenoside Rb2 SD, = 4, Student’s unpaired 0.05, ?? 0.01, ???? 0.0001, and ns (no significant difference) versus the control group; ## 0.01, #### 0.0001, and NS (no significant difference) versus the phloretin treatment group; 0.05, 0.0001, and (no significant difference) versus the Com C treatment group; 0.0001 Ginsenoside Rb2 versus the AICAR treatment group (mean SD, = 4). (b) The frequency of Th17 cells generated under Th17 polarization conditions. After na?ve CD4+ T cells were C1qdc2 activated, DMSO, 50? 0.0001 and ns versus the control group; # 0.05 and #### 0.0001 versus the phloretin treatment group; 0.0001 and versus the Com C treatment group; 0.0001 versus the AICAR treatment group (mean SD, = 4). (c) The frequency of Treg cells generated under Treg polarization conditions. After na?ve CD4+ T cells were activated, DMSO, 50? 0.01, ???? 0.0001, and ns versus the control group; #### 0.0001 and NS versus the phloretin treatment group; 0.001 and 0.0001 Ginsenoside Rb2 versus the Com C treatment group; 0.0001 versus the AICAR treatment group (mean SD, = 4). (aCc) One-way ANOVA followed by Tukey’s Ginsenoside Rb2 multiple comparison test was used in each statistical analysis. 4. Discussion Keeping appropriate immune homeostasis and self-tolerance is necessary for health. The anti-inflammatory effect of phloretin has been shown in animals and in vitro [12]. However, whether T cell immunity is influenced by phloretin is not completely clear. Therefore, we examined the impact and signaling mechanisms of.