Figure 4.
Ergosterol markedly decreased LPS-stimulated microglial activation and inflammation in ICR mice. (A) ICR mice were injected daily with Ergosterol (25 mg/kg, 50 mg/kg, i.p.) or vehicle (PBS, i.p.) for 5 days, followed by LPS (5 mg/kg, i.p.) or PBS injected for 24 h. The mice were then perfused and fixed. (B,C) An anti-IBA-1 antibody was used for immunohistochemistry. Quantification of IBA-1 fluorescence strength in the cerebral cortex or hippocampus (Nacl, n = 8 mice; LPS, n = 8 mice; Ergosterol 25 mg/kg + LPS is marked as ELT-L, n = 8 mice; Ergosterol 50 mg/kg + LPS is marked as ELT-H, n = 8 mice). (D,E) The mRNA levels of the IL-1β, IL-6, TNF-α, and INOS inflammatory factors in the cortex and hippocampus of the mice were measured with qRT-PCR. (F) The p-P65, IL-6, and IBA-1 protein levels were determined by Western blot. Actin was used as a housekeeping control (Nacl, n = 3 mice; LPS, n = 3 mice; ELT-L, n = 3 mice; ELT-H, n = 3 mice). (G) The relative expression level of protein to actin was calculated by densitometry. (H,I) The protein levels of IL-1β and IL-6 in plasma were detected by ELISA. (J) The expression levels of NO in plasma were detected with a Griess assay. Statistical significance was analyzed using one-way ANOVA and t test. # Significantly different from the control; * significantly different from the LPS-treated group. Significance: # p < 0.05, ## p < 0.01, ### p < 0.001, #### p < 0.0001, * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.
Figure 4.
Ergosterol markedly decreased LPS-stimulated microglial activation and inflammation in ICR mice. (A) ICR mice were injected daily with Ergosterol (25 mg/kg, 50 mg/kg, i.p.) or vehicle (PBS, i.p.) for 5 days, followed by LPS (5 mg/kg, i.p.) or PBS injected for 24 h. The mice were then perfused and fixed. (B,C) An anti-IBA-1 antibody was used for immunohistochemistry. Quantification of IBA-1 fluorescence strength in the cerebral cortex or hippocampus (Nacl, n = 8 mice; LPS, n = 8 mice; Ergosterol 25 mg/kg + LPS is marked as ELT-L, n = 8 mice; Ergosterol 50 mg/kg + LPS is marked as ELT-H, n = 8 mice). (D,E) The mRNA levels of the IL-1β, IL-6, TNF-α, and INOS inflammatory factors in the cortex and hippocampus of the mice were measured with qRT-PCR. (F) The p-P65, IL-6, and IBA-1 protein levels were determined by Western blot. Actin was used as a housekeeping control (Nacl, n = 3 mice; LPS, n = 3 mice; ELT-L, n = 3 mice; ELT-H, n = 3 mice). (G) The relative expression level of protein to actin was calculated by densitometry. (H,I) The protein levels of IL-1β and IL-6 in plasma were detected by ELISA. (J) The expression levels of NO in plasma were detected with a Griess assay. Statistical significance was analyzed using one-way ANOVA and t test. # Significantly different from the control; * significantly different from the LPS-treated group. Significance: # p < 0.05, ## p < 0.01, ### p < 0.001, #### p < 0.0001, * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.
Figure 5.
Ergosterol significantly reduced LPS-induced neuron injury in ICR mice. (A) The PSD95, Synapsin-1, and Synaptophysin protein levels in the cortex of mice were determined by Western blot. Actin was used as a housekeeping control (Nacl, n = 3 mice; LPS, n = 3 mice; ELT-L, n = 3 mice; ELT-H, n = 3 mice). (B) The densitometric method was used to estimate the relative expression of the protein compared to actin. (C,D) Immunohistochemistry was performed on anti-Synapsin-1 antibody. DAPI was used as a marker of the nucleus. Quantification of the relative fluorescence strength of Synapsin-1 in the region of the cortex or hippocampus was analyzed (Nacl, n = 8 mice; LPS, n = 8 mice; ELT-L, n = 8 mice; ELT-H, n = 8 mice). All data are mean ± SEM of triplicate values. Statistical significance was analyzed using one-way ANOVA and t test. # Significantly different from the control; * significantly different from the LPS-treated group. Significance: # p < 0.05, ## p < 0.01, ### p < 0.001, * p < 0.05, ** p < 0.01, and *** p < 0.001.
Figure 5.
Ergosterol significantly reduced LPS-induced neuron injury in ICR mice. (A) The PSD95, Synapsin-1, and Synaptophysin protein levels in the cortex of mice were determined by Western blot. Actin was used as a housekeeping control (Nacl, n = 3 mice; LPS, n = 3 mice; ELT-L, n = 3 mice; ELT-H, n = 3 mice). (B) The densitometric method was used to estimate the relative expression of the protein compared to actin. (C,D) Immunohistochemistry was performed on anti-Synapsin-1 antibody. DAPI was used as a marker of the nucleus. Quantification of the relative fluorescence strength of Synapsin-1 in the region of the cortex or hippocampus was analyzed (Nacl, n = 8 mice; LPS, n = 8 mice; ELT-L, n = 8 mice; ELT-H, n = 8 mice). All data are mean ± SEM of triplicate values. Statistical significance was analyzed using one-way ANOVA and t test. # Significantly different from the control; * significantly different from the LPS-treated group. Significance: # p < 0.05, ## p < 0.01, ### p < 0.001, * p < 0.05, ** p < 0.01, and *** p < 0.001.