Fig 1.
FB significantly reduced the volume of cerebral infarction and improved the behavioral indexes.
(A) protocol for an in vivo study. (B) Chemical structure of FB. (C) Representative brain slices stained with TTC. (D) Quantitative evaluation of infarct volume. (E) Zea-Longa score results. n = 8 per group. All plots are presented as the mean ± SD. ##p<0.01 vs. the sham group; *p<0.05, **p<0.01 vs. the MCAO/R group.
Fig 2.
FB reduced cerebral pathological injury of MCAO rats.
(A) The image positions corresponding to the cortex and the CA1 region of the hippocampus in each group. (B) Representative H&E and Nissl staining images in the cortex and the CA1 region of the hippocampus in each group. Scale bar = 50μm, n = 6. (C, D) Quantitative analysis of the H&E and Nissl staining. (E) Representative TEM images in the cortex and the CA1 region of the hippocampus in each group. n = 3. All plots are presented as the mean ± SD. ##p<0.01 vs. the sham group; **p<0.01 vs. the MCAO/R group.
Fig 3.
FB inhibited cell apoptosis in MCAO/R rats.
(A) Representative TUNEL staining images in the cortex and the CA1 region of the hippocampus in each group. Scale bar = 50μm, n = 6. (B, C) Relative density (% of sham) of the TUNEL staining. (D) Immunohistochemical analysis of Cl-caspase 3 in the cortex and the CA1 region of the hippocampus in each group. n = 6. (E) Quantification of Cl-caspase 3 expression. All plots are presented as the mean ± SD. ##p<0.01 vs. the sham group; **p<0.01 vs. the MCAO/R group.
Fig 4.
FB remitted the inflammation response and inhibited NLRP3 pathway in MCAO/R rats.
(A, B, C, D, E, and F) The levels of IL-1β, IL-18, MCP-1, IL-6, ICAM-1, and TNF-α in serum were measured using ELISA kit. n = 8. (G) Protein expression of NLRP3, IL-1β, and caspase-1 in ischemic penumbra of rat brains were measured using western blot analysis. n = 3. (H) Quantification of protein expression. (I-N) Immunohistochemical analysis of NLRP3, IL-1β, and NFкB in the cortex and the CA1 region of the hippocampus in each group. n = 6. All plots are presented as the mean ± SD. ##p<0.01 vs. the sham group; *p<0.01, **p<0.01 vs. the MCAO/R group.
Fig 5.
FB suppressed oxidative stress and increased Sirt1 expression in MCAO/R rats.
(A, B, C, D) Serum levels of SOD, GSH-Px, CAT, and MDA in rats were measured. n = 8. (E) Protein expression of Sirt1 in ischemic penumbra of rat brains were assayed using western blot analysis. n = 3. (F) Quantification of protein expression. All plots are presented as the mean ± SD. ##p<0.01 vs. the sham group; **p<0.01 vs. the MCAO/R group.
Fig 6.
Sirt1 knockdown abolished the inhibition of FB on inflammatory response and oxidative stress in MCAO/R rats.
(A) Representative brain slices stained by TTC. n = 6. (B) Quantitative evaluation of infarct volume. (C) Zea‑Longa score results. n = 8. (D) The levels of SOD, GSH-Px, CAT, and MDA in serum were measured using relevant reagent kits. n = 8. (E) The levels of IL-1β, IL-18, MCP-1, IL-6, ICAM-1, and TNF-a in serum of rats were measured by ELISA kits. n = 8. All plots are presented as the mean ± SD. #p<0.05, ##p<0.01 vs. the AAV-Cont group; **p<0.01 vs. the AAV-Cont+FB group.
Fig 7.
Sirt1 knockdown abolished the inhibition of FB on NLRP3 pathway.
(A) Expression of Sirt1, NLRP3, and IL-1β in ischemic penumbra of rat brains were assayed by western blot analysis. (B, C, and D) Quantification of protein expression. n = 3. All plots are presented as the mean ± SD. #p<0.05, ##p<0.01 vs. the AAV-Cont group; **p<0.01 vs. the AAV-Cont+FB group.
Fig 8.
Illustration of the pharmacological effect and possible underlying mechanism of FB in protecting against CIRI.