Fig 1.
Chemical structures of Curcumin and Ambrosin (A) and diagrammatic table of the experimental design (B).
Table 1.
Comparison of in silico predictions of pharmacokinetic properties of ambrosin and curcumin using different tools.
Fig 2.
Effect of different treatments on different parameters of MWM.
a) Effect on the mean escape latency time. b) Effect on the mean time spent in the target quadrant. Results are expressed as mean±SEM (n = 8). *Significant difference from normal group at p< 0.05. @Significant difference from control (LPS) group at p< 0.05.
Table 2.
Mean escape latency time in the MWM.
Fig 3.
Effect of different treatments on LPS-induced memory deterioration in mice utilizing the object recognition test.
a) Exploration time of familiar (F) vs. the novel object (N). b) Discrimination index (DI) Results are expressed as mean ±SEM (n = 8). Statistical analyses were carried out a) by using Student’s t-test, while in b) by one-way ANOVA followed by Tukey’s multiple comparison test. #Significant difference versus correspondent N group at p< 0.05. *Significant difference from normal group at p< 0.05. @Significant difference from control (LPS) group at p< 0.05.
Fig 4.
Suppression of NF-κβp65 transcript (a) and protein levels (b) by curcumin and ambrosin administration. Data represented mean ± SEM (n = 6). Statistical analysis was performed using one-way ANOVA followed by Tukey`s multiple comparison test. *Significantly different from normal group at P<0.05. @Significantly different from control group at P<0.05.
Fig 5.
Curcumin and ambrosin alleviated neuroinflammation by inhibition of repression of pro-inflammatory cytokines, viz., TNF-α (a), IL-1β and enzymes, viz., COX-2 (c). Data represented mean ± SEM (n = 6). Statistical analysis was performed using one-way ANOVA followed by Tukey`s multiple comparison test. *Significantly different from normal group at P<0.05. @Significantly different from control group at P<0.05.
Fig 6.
Curcumin and ambrosin abated the elevation in BACE1 levels after LPS administration.
Data represented mean ± SEM (n = 6). Statistical analysis was performed using one-way ANOVA followed by Tukey`s multiple comparison test. *Significantly different from normal group at P<0.05. @Significantly different from control group at P<0.05.
Fig 7.
Curcumin and ambrosin abated the elevation in iNOS levels after LPS administration.
Data represented as mean ± SEM (n = 6). Statistical analysis was performed using one-way ANOVA followed by Tukey`s multiple comparison test. *Significantly different from normal group at P<0.05. @Significantly different from control group at P<0.05.
Fig 8.
Histopathological lesions in the cerebral cortex of mice (magnifications: 10X, 40X).
The photomicrograph illustrates the following groups: (a, b) normal mice showing normal cerebral cortex, (c, d) control mice showing neuronal degeneration associated with presence of neurofibrillary tangles (arrow) and amyloid plaque with eosinophilic core, (e, f) Ambrosin (5 mg/kg/day) treated mice showing neurons bearing tangles and gliosis, (g, h) Ambrosin (10 mg/kg/day) treated mice demonstrating lower number of degenerated neurons, and (i, j) Curcumin treated mice showing scattered degenerated neurons. (Haematoxylin and eosin stain, (H&E stain)).
Fig 9.
Histopathological lesions in the hippocompus of mice (magnifications: 10X, 40X).
The photomicrograph illustrates the following groups: (a, b) normal mice showing normal hippocampal neurons, (c, d) control mice showing extensive degeneration of pyramidal neuronal cells, (e, f) Ambrosin (5 mg/kg/day) treated mice showing decreased number of degenerated neurons, (g, h) Ambrosin (10mg/kg/day) treated mice showing degeneration of individual pyramidal neurons, and (i, j) Curcumin treated mice showing normal neuronal cells. (Haematoxylin and eosin stain, (H&E stain)).
Fig 10.
Brain sections of mice stained with Congo red stain for the demonstration of amyloid plaques (magnifications: 4X, 10X, 40X).
The photomicrograph illustrates the following groups: (a-c) normal mice showing normal cerebral cortex (b) and hippocampus (c), (d-f) control mice showing abundant red-stained amyloid plaques in the cerebral cortex (e) and hippocampus (f), (g-i) Ambrosin (5mg/kg/day) treated mice showing decreased number of amyloid plaques in the cerebral cortical (h) and hippocampal tissues (i), (j-l) Ambrosin (10mg/kg/day) treated mice showing few amyloid plaques in cerebral cortex (k) and hippocampus (i),(m-o) Curcumin treated mice showing small amyloid plaques in cerebral cortex (n) and no amyloid plaques in hippocampus (o). (Congo red stain).
Table 3.
Quantification of surviving neurons in the hippocampal CA1 region and quantification of amyloid plaques found by histochemical examination of brains of tested groups.
Table 4.
Quantification of pro-inflammatory and pro-apoptotic markers traced by immunohistochemical examination of brains of tested groups.
Fig 11.
COX-2 immune-stained neurons in the brain sections of mice (magnifications: 10X, 40X).
The photomicrograph illustrates the following groups: (a, b) normal mice showing insignificant COX-2-immune positive cells, (c, d) control mice showing significant increase of COX-2 expression in the cerebral cortical neurons, (e, f) Ambrosin (5 mg/kg/day) treated mice showing reduction of COX-2 expression, (g, h) Ambrosin (10 mg/kg/day) treated mice showing COX-2 expression in fewer numbers of neurons, and (i, j) Curcumin treated mice showing individual COX-2 immune-stained cells. (COX-2 immunohistochemical stain).
Fig 12.
CD68 -immune reactive microglia in the brain sections of mice (magnifications: 10X, 40X).
The photomicrograph illustrates the following groups: (a, b) normal mice showing sparse weakly CD68 positive immune stained cells, (c, d) control mice showing significant increase of CD68 immune stained microglia surrounding amyloid plaques, (e, f) Ambrosin (5mg/kg/day) treated mice showing reduction of CD68 immune stained microglia, (g, h) Ambrosin (10mg/kg/day) treated mice showing small focal aggregates of CD68 immune stained microglia, and (i, j) Curcumin treated mice showing few CD68 immune stained microglia. (CD68 immunohistochemical stain).
Fig 13.
Cleaved caspase-3 immune reactive neurons in the brain sections of mice (magnifications: 10X, 40X).
The photomicrograph illustrates the following groups: (a, b) normal mice showing no caspase-3 immune reactive cells, (c, d) control mice showing significant increase of caspase-3 immune reactivity, (e, f) Ambrosin (5 mg/kg/day) treated mice showing reduction of caspase-3 immune reactivity, (g, h) Ambrosin (10 mg/kg/day) treated mice showing significant reduction of caspase-3 immune reactivity, and (i, j) Curcumin treated mice showing weak caspase-3 immune reactivity. (Cleaved caspase-3 immunohistochemical stain).
Fig 14.
Binding modes of ambrosin (a) and curcumin (b) showing polar interactions with DNA bases within NF-ҡβp65. DNA nucleotides are indicated by light green colors. Amino acids residues are blue pink in color. Hydrogen bonds are indicated as dotted arrows Hydrophobic interactions are highlighted in violet circles.
Table 5.
Docking Scores and interaction modes of Curcumin and Ambrosin with NF-ҡβp65 (PDB ID: 1VKX).