Fig. 3: Effects of pharmacological inhibition of hypothalamic inflammation on feeding behavior and expression of food intake-regulating neuropeptides in the diabetic mouse hypothalamus. | Nature Communications

Fig. 3: Effects of pharmacological inhibition of hypothalamic inflammation on feeding behavior and expression of food intake-regulating neuropeptides in the diabetic mouse hypothalamus.

From: Astrocytic pyruvate dehydrogenase kinase-2 is involved in hypothalamic inflammation in mouse models of diabetes

Fig. 3

Bay 11-7085 (an inhibitor of I-κB phosphorylation, 1 µg) and Ki20227 (an inhibitor of CSFR1, 1 µg) solution were administered via multiple icv injections starting from 4 d post-STZ injection to the following indicated time points. The schematic diagram represents the timeline of experimentation and the route of administration of inhibitors (a). Immunofluorescence analyses show the immunoreactivity of NF-κB p65 and Iba-1 (b) in the hypothalamus at 3 w post-vehicle/STZ injection. Inserts show the translocation of NF-κB p65 in DAPI-positive nuclei. Quantification of the percentage of NF-κB p65-positive nuclei (###p = 9.3E-5, STZ + Bay11-7085) and the total number of Iba-1-positive microglia (###p = 2.5E-7, STZ + Ki20227) per mm2 are shown in the adjacent images. Microscope data were gathered using six randomly selected fields captured at the same magnification. Scale bars indicate 100 µm (left) and 200 µm (right). The expression of proinflammatory cytokines such as Tnf-α (###p = 6.7E-8, STZ + Bay11-7085; and ###p = 2.5E-7, STZ + Ki20227), Il-1β (###p = 3.7E-5, STZ + Bay11-7085; and ###p = 0.0008, STZ + Ki20227), and Il-6 (#p = 0.0120, STZ + Bay11-7085; and ##p = 0.0010, STZ + Ki20227) was assessed by real-time RT-PCR (c). Food intake (*p = 0.0160, STZ + Bay11-7085; and §p = 0.0323, STZ + Ki20227 at 28 d) was measured (d), and the expression of Npy (###p = 0.0006, STZ + Bay11-7085; and ###p = 0.0002, STZ + Ki20227), Agrp (###p = 0.0003, STZ + Bay11-7085; and ###p = 0.0004, STZ + Ki20227), and Pomc (###p = 0.0001, STZ + Bay11-7085; and #p = 0.0110, STZ + Ki20227) mRNA was assessed by real-time RT-PCR (e). Results for mRNA expression are displayed as the fold increase of gene expression normalized to Gapdh. * or §p < 0.05 or **p < 0.01 or ***p < 0.001 versus WT/KO control animals (b, c, e) or STZ-injected animals (d). One-way ANOVA (b, c, e) and two-way ANOVA with Tukey’s post hoc test (d), n = 5 (b), n = 6 (d), and n = 4 (c, e); mean ± SEM. Value of “n” indicates the number of animals. Source data are provided as a Source data file. d day(s); STZ streptozotocin; Bay Bay 11-7085; Ki Ki20227.

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