Metabolite analysis reveals elevated lactate and glucose metabolism in 5-day post-SE hippocampal metabolism—Signatures of warburg metabolism.

(A) Representative ¹H NMR spectra of hippocampal metabolites. Red: Control, Black: Pilocarpine, Green: Difference spectrum (scaled 4x). The NMR peaks corresponding to significantly changed levels of metabolites are labeled. A non-significant resonance at 3.716 ppm was removed for clarity. Non-standard abbreviations used are Suc, succinate; GPC, sn-glycero-3-phosphocholine; Myo, myo-inositol; OPC, O-phosphocholine; GABA, 4-aminobutyrate; NAA, N-acetylaspartate. (B) Partial least squares analysis demonstrates significant difference between control and 5-day post-SE metabolism. The metabolites showed significant separation (>95% confidence) between control mice (red) and pilocarpine-treated mice (green). (C) Metabolite changes correlated and anti-correlated with lactate in control and 5-day post-SE metabolism. The top 25 metabolites pro- and anti-correlated with lactate are shown along with their correlation coefficients calculated in Metaboanalyst 4.0 [62, 63]. (D) Representative ¹H-¹³C HSQC NMR spectrum used for identification of hippocampal metabolites after infusion of [U-¹³C]-glucose and administration of pilocarpine. The NMR peaks corresponding to significantly changed levels of metabolites are circled and labeled. Abbreviations used: Suc, succinate; GPC, sn-glycero-3-phosphocholine; Myo, myo-inositol; GABA, 4-aminobutyrate; NAA, N-acetylaspartate. (E) Quantitative RT-PCR of Pyruvate Carboxylase (PC) expression levels in hippocampus at day 5 following SE with pilocarpine (Pilo) or control (saline). (n = 4–6 mice/group). Data are represented as mean ± SEM. P >0.05, by unpaired student’s t-test using Prism 9.0 (Graphpad). Refer to supplemental materials for more details on the number of replicates and statistical analyses for each experiment.

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