Fig. 5

SP administration reverses MGs’ aging phenotype by reducing the level of succinylation. A. Schematic illustration of (a) the generation of SCoA via KGDHC and the metabolite changes follow the SP treatment, (b) Schematic workflow of the experiment of SP pretreatment and the following experiments. B. Succinylation level of LPS treatment with or without SP pretreatment in BV2 cells were detected by WB. C. The concentration of SCoA in cellular total lysate of LPS treatment with or without SP pretreatment in BV2 cells were detected by colorimetric method. D. Schematic workflow of detecting the senescence MG indicators after SP pretreatment. E. mtROS generation was detected by MitoSox staining after the SP pretreatment in the physiological and inflammatory condition; (a) mtROS generation was observed and detected by fluorescence microscopy and the MFI was measured. Bar = 50 μm; (b) mtROS generation of BV2 cells treated with SP and LPS was detected by FC. F. Lipid accumulation was detected by Nile Red staining after the SP pretreatment in the physiological and inflammatory condition; (a) Lipid accumulation was observed and detected by fluorescence microscopy and the MFI was measured. Bar = 50 μm; (b) Lipid accumulation of BV2 cells treated with SP and LPS was detected by FC. G. MDA concentration of BV2 cells treated with SP and LPS was detected by colorimetric method. H. Pro-inflammatory cytokines transcription level of Il-1β, Il-6 and Tnf-α in the, (a) PriMGs and (b) BV2 cells treated with SP and LPS were detected by qPCR. I. Schematic workflow of SP pretreatment in the VP16 induced aging model. J. SA-β-Gal positive cells were counted and calculated following the pretreatment of SP in the VP16 induced aging model. Bar = 50 μm K. Schematic workflow of SP pretreatment in the in vivo neuroinflammation model. L. The succinylation level of the hippocampus mito lysate of the in vivo neuroinflammation model pretreated with SP was detected by WB. n = 3/group, * P < 0.05, ** P < 0.01, *** P < 0.005, **** P < 0.001