Journal: Advanced Science

Article Title: H4K12 Lactylation Impairs Dopaminergic Neuron Energy Metabolism via the HIF‐1α/NDUFS1 Pathway in Parkinson's Disease

doi: 10.1002/advs.78220

Figure Lengend Snippet: H4K12la regulates HIF‐1α expression in PD models, and inhibition of lactylation by 2‐DG ameliorates related pathologies. (A) Western blotting analysis of H4K12la and HIF‐1α protein levels in SN4741 cells treated with 0.5 mM MPP + , NaLa, Lac, or NaCl for 24 h (N = 3). (B) Representative immunofluorescence images of SN4741 cells showing H4K12la (green) and HIF‐1α (red) expression in the nucleus under the indicated treatments. DAPI (blue) stains nuclei. Scale bar, 100 µm. Insets show 2 × magnifications of the boxed regions. (C) Quantitative analysis of the relative fluorescence intensity of H4K12la and HIF‐1α. (D,E) Western blotting analysis and quantification of H4K12la and HIF‐1α protein levels in the (D) striatum and (E) substantia nigra of MPTP‐induced PD mice (n = 3). (F, G) RT‐qPCR analysis of HIF‐1α mRNA levels in the (F) striatum and (G) substantia nigra of MPTP‐induced PD mice (n = 3). (H,I) Cells transfected with H4‐WT or H4‐K12R were treated with MPP + . H4K12la and HIF‐1α protein levels were assessed by Western blotting (N = 3). (J) Intracellular lactate levels in MPP + ‐treated SN4741 cells with or without 2‐DG treatment (N = 3). (K) Western blotting analysis of H4K12la and HIF‐1α protein levels in MPP + ‐treated SN4741 cells with or without 2‐DG co‐treatment (N = 3). (L) Representative immunofluorescence images of SN4741 cells showing that 2‐DG co‐treatment reduces MPP + ‐treated elevations of H4K12la (green) and HIF‐1α (red). Scale bar, 100 µm. (M) Corresponding quantification of fluorescence intensity for H4K12la and HIF‐1α. (N,O) Lactate levels in the (N) substantia nigra and (O) striatum homogenates of MPTP‐induced PD mice with or without 2‐DG treatment (n = 3). (P,Q) Western blotting analysis and quantification of H4K12la and HIF‐1α protein levels in the (P) striatum and (Q) substantia nigra of MPTP‐induced PD mice with or without 2‐DG treatment (n = 3). (R) Representative immunofluorescence images of midbrain sections from MPTP‐induced PD mice with or without 2‐DG treatment, showing TH (red) and H4K12la (green) expression. Scale bar, 200 µm (n = 3). Data are presented as mean ± SEM. ns, not significant, * p < 0.05, ** p < 0.01, and *** p < 0.001. Statistical significance was determined by one‐way ANOVA or Student's t‐test as appropriate. Lac, lactate; NaLa, sodium lactate; NaCl, sodium chloride; 2‐DG, 2‐deoxy‐D‐glucose. “n” refers to the number of mice per group. “N” refers to the number of biologically independent experiments.

Article Snippet: Mice in the 2‐DG + MPTP group were pretreated with 2‐DG (400 mg/kg, MCE, #HY‐13966) via intraperitoneal injection once daily for 3 days prior to MPTP administration.

Techniques: Expressing, Inhibition, Western Blot, Analysis, Immunofluorescence, Fluorescence, Quantitative RT-PCR, Transfection

Journal: Advanced Science

Article Title: H4K12 Lactylation Impairs Dopaminergic Neuron Energy Metabolism via the HIF‐1α/NDUFS1 Pathway in Parkinson's Disease

doi: 10.1002/advs.78220

Figure Lengend Snippet: 2‐DG ameliorates energy metabolism deficits, provides neuroprotection, and improves motor function in PD models by regulating H4K12la. (A,B) Measurement of (A) the ATP levels and (B) NAD + /NADH ratio in SN4741 cells treated with MPP + , with or without 2‐DG (N = 3). (C) Experimental schedule. (D–G) Measurement of the ATP levels and NAD + /NADH ratio in the (D,E) striatum and (F,G) substantia nigra of MPTP‐induced mice treated with or without 2‐DG (n = 3). (H) Representative immunohistochemical images and quantitative analysis of TH expression in the midbrain of mice treated with MPTP, with or without 2‐DG (n = 3). Scale bar, 200 µm. (I,J) Western blotting analysis and quantification of ATPB protein levels in the (I) striatum and (J) substantia nigra of mice treated with MPTP, with or without 2‐DG (n = 3). (K) RT‐qPCR analysis of TH mRNA levels in the substantia nigra of mice treated with MPTP, with or without 2‐DG (n = 3). (L–N) Behavioral assessments of MPTP‐induced mice with or without 2‐DG treatment: (L) time in the pole climbing test. (M) open‐field test showing representative images of mouse movement trajectories and total distance traveled. (N) Scores in the grasping test. n = 7 per group. Data are presented as mean ± SEM. * p < 0.05, ** p < 0.01, and *** p < 0.001. Statistical significance was determined by one‐way ANOVA or Student's t‐test as appropriate. ATPB, ATP synthase subunit beta. “n” refers to the number of mice per group. “N” refers to the number of biologically independent experiments.

Article Snippet: Mice in the 2‐DG + MPTP group were pretreated with 2‐DG (400 mg/kg, MCE, #HY‐13966) via intraperitoneal injection once daily for 3 days prior to MPTP administration.

Techniques: Immunohistochemical staining, Analysis, Expressing, Western Blot, Quantitative RT-PCR

Journal: CNS Neuroscience & Therapeutics

Article Title: Histone H3K18 Lactylation Contributes to Perioperative Neurocognitive Disorder Through Immune Checkpoint Lymphocyte Activation Gene 3 Mediated Microglial Pyroptosis

doi: 10.1002/cns.71058

Figure Lengend Snippet: The target genes regulated by H3K18la were identified by CUT&Tag assay combined with RNA‐seq. (A) Average plot and heat map displaying the binding density of H3K18la with different H3K18la binding peaks in hippocampus of the Sur group and Con group. (B) Genome‐wide distribution of the upregulated H3K18la‐binding peaks in mice after surgery. (C) Bar graph illustrating the number of target genes associated with upregulated and downregulated H3K18la peaks, and the top 2 enriched de novo motifs of target genes. (D) GO analysis of candidate target genes associated with upregulated H3K18la peaks. (E) Volcano plot of differentially expressed genes in RNA‐seq ( n = 3). (F) Heat map showing TOP 20 differential genes after 2‐DG treatment in RNA‐seq ( n = 3). (G) Venn diagram showing the intersection of H3K18la up‐regulated target genes and 2‐DG down‐regulated target genes. (H) Genome browser tracks show H3K18la binding at representative target gene loci. Peaks specifically located at the Lag3 promoters are identified by red rectangles. (I) H3K18la occupancy analysis by ChIP‐qPCR ( n = 3). Data are mean ± SD. Statistical significance was assessed by unpaired 2‐tailed Student t ‐test compared with Con group. **** p < 0.0001.

Article Snippet: Sur + 2‐DG mice received 2‐DG (250 mg/kg; MedChemExpress, HY‐13966) [ ] intraperitoneally once daily for 2 days preoperatively until 30 min before surgery.

Techniques: RNA Sequencing, Binding Assay, Genome Wide, ChIP-qPCR

Journal: CNS Neuroscience & Therapeutics

Article Title: Histone H3K18 Lactylation Contributes to Perioperative Neurocognitive Disorder Through Immune Checkpoint Lymphocyte Activation Gene 3 Mediated Microglial Pyroptosis

doi: 10.1002/cns.71058

Figure Lengend Snippet: H3K18la promotes Lag3 expression in microglial cells induced by either lactate or LPS and IFN‐γ. (A) BV2 cells were first treated with 0‐, 1‐, 5‐, and 25‐mM lactate for 24 h. Subsequently, cells pretreated with 25 mM lactate or 10 mM 2‐DG for 3 h were stimulated with LPS and IFN‐γ for 24 h. (B) Representative immunoblotting of Pan‐Kla, H3K18la and Lag3 in BV2 cells intervention with lactate, with histone H3 or GAPDH used for normalization. (C‐E) Quantitation of Pan‐Kla, H3K18la and Lag3 gray value ( n = 3). (F) The lactate levels of BV2 cells activated by LPS and IFN‐γ ( n = 5–6). (G) Representative immunoblotting of Pan‐Kla, H3K18la and Lag3 in BV2 cells activated by LPS and IFN‐γ, with histone H3 or GAPDH used for normalization. (H–J) Quantitation of Pan‐Kla, H3K18la and Lag3 gray value ( n = 3–4). (K) Representative fluorescence images of Pan‐Kla or Lag3 in BV2 cells. (L, M) The relative fluorescence intensities of Pan and Lag3 in BV2 cells ( n = 6). The images include scale indicators of 50 μm for size reference. The data are presented as the mean ± SD, A–D were assessed by unpaired 2‐tailed Student t ‐test compared with Con group. The other results were assessed by one‐way ANOVA followed by Tukey's post hoc test. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Sur + 2‐DG mice received 2‐DG (250 mg/kg; MedChemExpress, HY‐13966) [ ] intraperitoneally once daily for 2 days preoperatively until 30 min before surgery.

Techniques: Expressing, Western Blot, Quantitation Assay, Fluorescence