Lipid Isolated from a Leishmania donovani Strain Reduces Escherichia coli Induced Sepsis in Mice through Inhibition of Inflammatory Responses

Figure 3

Effect of LTL on expression levels of (a) NF-𝜅B p65 in peritoneal macrophage cells after 4 h incubation at concentration LTLd₂ (100 μg/mL) in the presence and absence of heat-killed E. coli (O18:K1; 1×10⁸ CFU/mL), from immunofluorescence level measured by confocal laser scanning microscopy. (b) ELISA results showed the nuclear extract of NF-𝜅B p65 in peritoneal macrophage cells after 1, 2, and 4 h incubation with LTLd₁ and LTLd₂. (c) Western blot analysis revealed nuclear and cytosolic NF-𝜅B p65 levels and p-I𝜅B protein expression level after treatment with LTLd₁ and LTLd₂ following 12 h incubation. (d) Immunofluorescence expression measured by confocal laser scanning microscopy (magnification 600x) and (e) by flow cytometry showing levels of CD14 and TLR4 in peritoneal macrophage cells after 4 h incubation at concentration LTLd₂ in the presence and absence of heat-killed E. coli (O18:K1; 1×10⁸ CFU/mL). The data are reported as the mean ± SEM of triplicate experiments (^∗𝑃<0.05, ^∗∗𝑃<0.01).