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. 2022 Jul 14;13:885685. doi: 10.3389/fimmu.2022.885685

Figure 5.

Figure 5

C646 blocked crotonyl-CoA induced macrophage activation and the upregulation of inflammatory factors and chemokine in TG. (A) C646 suppressed the upregulation of Iba1 mRNA induced by Crotonyl-CoA Lithium in TG. ** P < 0.01, n = 5-6, One-way ANOVA followed by Bonferroni’s test. (B–F) Representative images and Quantitative analysis of immunofluorescence staining showing C646 (1 μg) significantly decreased Crotonyl-CoA Lithium induced IBA-1 upregulation (E) and the number of IBA-1 positive cells (F) in TG. ** P < 0.01, *** P < 0.001, ****P < 0.001, One-way ANOVA followed by Bonferroni’s test, n = 3 mice per group, 3 sections per mouse. (G) The mRNA levels of Tnfα, Il1β and Il6 increased significantly after Crotonyl-CoA Lithium treatment, ** P < 0.01, *** P < 0.001, ****P < 0.001, n = 6, student’s t-test. (H) C646 significantly decreased the mRNA expression of Tnfα, Il1β and Il6 upregulated by Crotonyl-CoA Lithium, ** P < 0.01, *** P < 0.001, compared with the Crotonyl-CoA Lithium group, n = 6, student’s t-test. (I) Crotonyl-CoA Lithium significantly increased the mRNA level of Ccl2 and Ccl7, ** P < 0.01, *** P < 0.001, compared with the PBS group, n = 6-8, student’s t-test. (J) C646 significantly decreased the mRNA expression of Ccl2, Ccl7 and Cxcl10, *** P < 0.001, ****P < 0.0001, compared to Crotonyl-CoA Lithium, n = 6, student’s t-test. (K–N) Representative images (K, L) and statistics of the number of KCR and IBA-1 double-positive cells (M) and the fluorescence intensity of KCR in IBA-1 positive cells (N) 6 hours after intra-TG injection of PBS or Crotonyl-CoA Lithium on naïve mice. *P < 0.05, **P < 0.01, N = 3 mice per group, 3 sections per mice, student’s t-test. (O, P) Minocycline (1 nmol per mouse) significantly attenuated Crotonyl-CoA-induced mechanical allodynia when minocycline and Crotonyl-CoA Lithium were injected simultaneously, **P < 0.01, ****P < 0.0001 compared with the Crotonyl-CoA Lithium group, n = 8 in each group, Two-way RM ANOVA followed by Bonferroni’s test.