Naomaitai Ameliorated Brain Damage in Rats with Vascular Dementia by PI3K/PDK1/AKT Signaling Pathway
PI3K inhibitor reverses the therapeutic effect of NMT on VD rats. Bederson scoring system was used to detect neurological impairment in rats. MWM test was conducted to determine the spatial cognition and memory ability of rats. HE staining was utilized to observe the changes of brain tissue morphology in rats. ELISA was performed to determine the expression levels of brain damage markers, oxidative stress factors and inflammation related factors. Apoptosis-positive cells were determined via Tunel assays. Apoptosis-related protein expression was determined via Western blot assay. Immunofluorescence method and western blot assay were used to determine the protein expression of PI3K, PDK1, and AKT. LFB staining was utilized to observe the damage of white matter nerve fibers and the loss of myelin sheath. Immunofluorescence method was used to determine the MBP expression in white matter of rats. (a) The neurological deficit score and Morris water maze test; (b) hematoxylin & eosin staining (bar, 50μm); (c) ELISA was performed to determine the expression levels of NSE and S-100; (d) the expression levels of oxidative stress factors (MDA and SOD) in white matter; (e) the expression levels of inflammation related factors (IL-1, IL-6, TNF-, and IL-10) contents in white matter; (f) Tunel assays (bar, 50μm); (g) the protein expression of PI3K, PDK1 and AKT determined by Western blot assay; (h) the protein expression of PDK1 and Akt determined by Immunofluorescence method (bar, 50μm); (i) LFB staining (bar, 50μm); (j) the expression of MBP determined by Immunofluorescence method (bar, 50μm). $P<0.05 vs. HNMT group.
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