Fig 1.
The cytotoxicity of acrolein and H2O2 in FM3A cells.
The viable cell number of FM3A cells that were treated with PI EtOH extract at 5 μg/ml plus acrolein at 5 μM (A) or 2 μM (B) or 100 μM H2O2 and cultured for 24 and 48 hrs. The number of cells was counted with 0.25% trypan blue. Each value represents the mean ± SD (n = 3).
Fig 2.
The effect of P. igniarius and P. linteus extracts on the cytotoxicity of acrolein and iodoacetic acid.
The viable cell number of Neuro-2a cells that were treated with PI (A) or PL (B) DDW crude extract at 20, 50, 100 and 200 μg/ml after a pretreatment with 8 μM acrolein for 4 hrs. PI (C) and PL (D) DDW crude extract at 20, 50, 100 and 200 μg/ml after a pretreatment with 5 μM iodoacetic acid for 4 hrs. PL EtOH crude extract (E) at 2, 5, 10, 20 and 50 μg/ml after a pretreatment with 5 μM iodoacetic acid for 4 hrs. Cell viability was determined by MTT assay after 24 hrs. Each value represents the mean ± SD (n = 3).
Fig 3.
Effect of pretreatment of Neuro-2a cells with PL and PI extracts on cytotoxicity of acrolein.
The viable cell number of Neuro-2a cells that were treated with PI EtOH crude extract (A) at 2, 5, 10, 20 and 50 μg/ml, PI (B) or PL (C) DDW crude extract at 20, 50, 100 and 200 μg/ml for 4 hrs and treated with 8 μM acrolein. Cell viability was determined by MTT assay after 24 hrs. Each value represents the mean ± SD (n = 3).
Fig 4.
Matrix assay of ethanol extract.
EtOH extract dissolved in DMSO at 0.5, 1 and 2 μg/ml with acrolein at 2 μM (A); glucans of PI at 20 and 50 μg/ml and polysaccharides of PL at 10, 20 and 50 μg/ml with acrolein at 2 μM (B); PI ethanol extract dissolved in DDW at 10, 20 and 50 μg/ml with acrolein at 2 μM (C) or 5 μM (D). Cell viability was determined by MTT assay after 12, 24 and 48 hrs. Each value represents the mean ± SD (n = 3).
Fig 5.
The effect of PIDMSO on a mouse stroke model.
The effect of PIDMSO on brain infarction size (A), the level of PC-Acro in plasma (B) and the formation of FDP-lys from acrolein in protein (C). Experiments were performed using 6 mice in each group as described in Materials and Methods. Data are shown as mean ± SE. ***P<0.001 compared with control mice.
Fig 6.
The effect of 200 ng/kg and 2 μg/kg PIDMSO on brain infarction size.
Fig 7.
The levels of PC-Acro and albumin in brain tissue.
Total proteins (5 μg) from brain tissues at the locus of infarction (A,B) and a normal area (C,D) were stained with Coomassie Brilliant Blue R250 (A,C), and the relative levels of PC-Acro and albumin were estimated by Western blotting using antibodies against mouse albumin (B,D,E). The results represent the mean ± SD of 6 mice.
Fig 8.
Total ion current (TIC) chromatogram of polyphenols from Phellinus igniarius.
Table 1.
Tentative identification of polyphenol extract from Phellinus igniarius by UPLC/ESI-MS in negative.
Table 2.
Compounds identified in polyphenol extract from Phellinus igniarius by assignment of their proton resonances.
Fig 9.
Structure of precursor polyphenols.