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. 2021 Oct 20;12:710513. doi: 10.3389/fimmu.2021.710513

Figure 6.

Figure 6

Microbiota ablation with antibiotics eliminated the protective effects of ESPs on cognitive impairment, neuroinflammation, and synapse in HF diet-induced obese mice. (A) The nesting behavior test (n =12). (B) The object location test (n =12). (C) The Y-maze memory test (n =12). (D) The novel object recognition test (n =12). (E) The temporal order memory test (n =12). (F) Representative immunofluorescent staining and quantification of Iba-1+ cells in the CA1, CA3, and DG regions of HIP (n = 3, 2 images per mouse, scale bar 50 μm). The image captured from the box was marked with a solid line (scale bar 10 μm). (G, H) The mRNA expression levels of CD68 and proinflammatory cytokines, including TNF-α, IL-1β, and IL-6 in the HIP (n = 6). The relative mRNA levels were normalized with reference gene (β-actin). (I) The protein expression levels of TNF-α in the HIP (n = 8). (J) Immunofluorescent staining and quantification of Iba-1+ cells in the PFC (n = 3, 2 images per mouse, scale bar 50 μm). (K, L) The mRNA expression levels of CD68 and proinflammatory cytokines, including TNF-α, IL-1β, and IL-6 in the PFC (n = 6). The relative mRNA levels were normalized with reference gene (β-actin). (M) The protein expression levels of TNF-α in the PFC (n = 8). (N) Representative ultrastructure of synapses in the CA1 region of mice on the electron micrograph (scale bar 100 nm) and image analysis of the length of active zone, thickness of PSD, width of the synaptic cleft, and the synaptic curvature (n = 3, 6 images per mouse). (O–Q) The protein expression levels of BDNF, SYN and PSD-95 in the HIP (n = 8). (R–T) The protein expression levels of BDNF, SYN and PSD-95 in the PFC (n = 8). Values are represented as mean ± SEM.