Activation of Central Alpha 7 Nicotinic Acetylcholine Receptor Reverses Suppressed Immune Function of T Lymphocytes and Protects Against Sepsis Lethality

Figures

• 

  Figure 1. ICV injection of PNU282987 ameliorated sepsis induced multiple organ dysfunction. (1A) Histopathologic changes of heart, liver, lungs, and kidneys were measured by HE staining (×100 magnification). (1B, 1C, and 1E) Serum biochemical parameters, including creatine kinase (CK), CK-MB, aspartate aminotransferase (AST), alanine aminotransferase (ALT), cholylglycine (CG), blood urea nitrogen (BUN), and creatinine (Cr), were quantified by HITACHI 7600 to reflect organ dysfunction, respectively (n=6, *P<0.01 as compared with the sham groups; #P<0.01 as compared with the sepsis groups; **P<0.01 as compared with other dose groups). (1D) Wet to dry ratio (W/D) and myeloperoxidase (MPO) activity of pulmonary tissues were quantified by weighting and ELISA kit, respectively (n=6, *P<0.01 as compared with the sham groups; #P<0.01 as compared with the sepsis groups; **P<0.01 as compared with other dose groups).

• 

  Figure 2. ICV injection of MLA exacerbated multiple organ injury in septic rats. (2A) Histopathologic alterations of heart, liver, lungs, and, kidneys were determined by HE staining (×100 magnification). (2B, 2C, and 2E) Serum biochemical parameters, including CK, CK-MB, AST, ALT, CG, BUN, and Cr, were quantified by HITACHI 7600 to reflect organ dysfunction, respectively (n=6, *P<0.01 as compared with the sham groups; #P<0.01 as compared with the sepsis groups). (2D) W/D ratio and MPO activity of pulmonary tissues were quantified by weighting and ELISA kit, respectively (n=6, *P<0.01 as compared with the sham groups; #P<0.01 as compared with the sepsis groups).

• 

  Figure 3. The time-dependent response of PNU282987 and MLA on sepsis induced multiple organ dysfunction. (3A, 3B, and 3D) Serum biochemical parameters, including CK, CK-MB, AST, ALT, CG, BUN, and Cr, were quantified by HITACHI 7600 to reflect organ dysfunction, respectively. (3C) W/D ratio and MPO activity of pulmonary tissues were quantified by weighting and ELISA kit, respectively (n=6, *P<0.01 as compared with the sham groups at the identical time points; #P<0.01 as compared with the sepsis groups at the identical time points; **P<0.01 as compared with sepsis groups at the identical time points; &P<0.01 as compared with different time points with PNU282987 administration; ##P<0.01 as compared with different time points with MLA administration).

• 

  Figure 4. Effects of treatment with PNU282987 on proliferation and secretion of splenic CD4+ T cells in septic rats. (4A) The proliferative activity of CD4+ T cells was measured by CCK8 assay and quantified by ELISA plate reader at 450 nm. (4B) Levels of IL-2, IL-4, and IFN-γ in culture supernatants were determined by ELISA kits. (4C) The mRNA expressions of IL-2, IL-4, and IFN-γ in CD4+ T cells were quantified by quantitative real-time PCR (n=6, *P<0.01 as compared with the sham groups; #P<0.01 as compared with the sepsis groups; **P<0.01 as compared with other dose groups).

• 

  Figure 5. Effects of treatment with MLA on proliferation and secretion of splenic CD4+ T cells in rats following septic challenge. (5A) The proliferative activity of CD4+ T cells was measured by CCK8 assay and quantified by ELISA plate reader at 450 nm. (5B) Levels of IL-2, IL-4, and IFN-γ in culture supernatants were determined by ELISA kits. (5C) The mRNA expressions of IL-2, IL-4, and IFN-γ in CD4+ T cells were quantified by quantitative real-time PCR (n=6, *P<0.01 as compared with the sham groups; #P<0.01 as compared with the sepsis groups).

• 

  Figure 6. The time-dependent response of PNU282987 and MLA on the immune function of splenic CD4+ T cells in rats following septic challenge. (6A) The proliferative activity of CD4+ T cells was measured by CCK8 assay and compared among different time points after CLP surgery. (6B) Levels of IL-2, IL-4, and IFN-γ in culture supernatants were measured by ELISA kits. (6C) The mRNA expressions of IL-2, IL-4, and IFN-γ in CD4+ T cells were quantified by quantitative real-time PCR (n=6, *P<0.01 as compared with the sham groups at the identical time points; #P<0.01 as compared with the sepsis groups at the identical time points; **P<0.01 as compared with the sepsis groups at the identical time points; &P<0.01 as compared with different time points with PNU282987 administration; ##P<0.01 as compared with different time points with MLA administration).

• 

  Figure 7. Effects of treatment with PNU282987 and MLA on the survival rates of septic rats. The survival rates were recorded at 12 h, 24 h, 36 h, 48 h, 72 h, 96 h, 120 h, 144 h, and 168 h post CLP operation, respectively. ICV injection of PNU282987 significantly increased the survival rate of septic rats in comparison to that in the sepsis group (n=26, 80% vs. 40%, P<0.05). No remarkable difference was found between the sepsis with MLA injection group and the sepsis group.