Fig. 6: mGluR5-induced NOX2 activation in Kupffer cells occurs via a PKC-dependent pathway. | Nature Communications

Fig. 6: mGluR5-induced NOX2 activation in Kupffer cells occurs via a PKC-dependent pathway.

From: Binge drinking triggers VGLUT3-mediated glutamate secretion and subsequent hepatic inflammation by activating mGluR5/NOX2 in Kupffer cells

Fig. 6

a Measurement of serum ALT levels in KC from myeloid cell-specific Cybb knockout mice (KCΔCybb and MyelΔCybb) fed with 2-week EtOH plus binge drinking (n = 6/group). b Representative H&E stains in liver sections. Scale bar, 50 μm. c Representative flow cytometry panels (left) and bar graphs (right) showing frequencies of neutrophils in liver MNCs from Cybb knockout mice (KCΔCybb and MyelΔCybb) compared to floxed mice (n = 6/group). d mRNA expression levels of Cxcl1, Il1b and Tnf in isolated KCs (n = 4/group). e Generation of conditioned media (CM) by co-culturing WT HEPs with KCΔfl/fl or KC ΔCybb with 50 mM EtOH for 24 h, followed by culturing isolated neutrophils with CM for 12 h and analyzing Tnf, Il1b, and Elane mRNA expression (n = 4/group). f Relative mRNA expressions of Prkca, Prkcb, and Prckg in isolated KCs from KCΔCybb and KCΔGrm5 mice (n = 4/group). g Gene ontology analysis and mRNA expression of protein kinase C (PKC)-related genes in KCs from Myelfl/fl and MyelΔCybb mice. Green dots indicate the PKC-related gene sets. h mRNA expression of Prkca, Prkcb, and Prckg in isolated KCs treated with 100 μM MSG (30 min) or 10 μM CHPG (10 min) (n = 4 /group). i Measurement of ALT levels following intraperitoneal injection of VEH or PKC inhibitor (PKCi; chelerythrine chloride; 10 mg/kg) for 3 consecutive days before sacrifice (n = 7/group). j, k Representative of ROS generation and mRNA expression in isolated KCs from VEH or PKCi-treated mice (n = 4/group). l H&E and immunostaining of ALDH4A1 and CYP2E1 in liver sections. Scale bar, 50 μm. m Western blot analysis for the downstream signaling pathways of mGluR5 in isolated KCs from KCfl/fl, KCΔGrm5, and KCΔCybb mice. Statistical comparisons were made using one-way ANOVA with Tukey’s multiple comparisons test (e), multiple comparison by two-stage step-up method of Benjamini, Krieger and Yekutieli. F (d, f, h), or unpaired t-test (a, c, i, k). Data are presented as mean ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001. Source data are provided as a Source Data file.

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