Genistein improves 3-NPA-induced memory impairment in ovariectomized rats: impact of its antioxidant, anti-inflammatory and acetylcholinesterase modulatory properties.

Huntington's disease (HD) is a progressive neurodegenerative disorder. The pre-motor symptomatic stages of the disease are commonly characterized by cognitive problems including memory loss. 3-Nitropropionic acid (3-NPA) is a mitochondrial toxin that produces selective lesions in the brain similar to that of HD and was proven to cause memory impairment in rodents. Phytoestrogens have well-established neuroprotective and memory enhancing effects with fewer side effects in comparison to estrogens. This study investigated the potential neuroprotective and memory enhancing effect of genistein (5, 10 and 20 mg/kg), a phytoestrogen, in ovariectomized rats challenged with 3-NPA (20 mg/kg). These potential effects were compared to those of 17β-estradiol (2.5 mg/kg). Systemic administration of 3-NPA for 4 consecutive days impaired locomotor activity, decreased retention latencies in the passive avoidance task, decreased striatal, cortical and hippocampal ATP levels, increased oxidative stress, acetylcholinesterase (AChE) activity, cycloxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) expressions. Pretreatment with genistein and 17β-estradiol attenuated locomotor hypoactivity, increased retention latencies in the passive avoidance task, increased ATP levels, improved the oxidative stress profile, attenuated the increase in AChE activity and decreased the expression of COX-2 and iNOS. Overall, the higher genistein dose (20 mg/kg) was the most effective. In conclusion, this study suggests neuroprotective and memory enhancing effects for genistein in a rat model of HD. These effects might be attributed to its antioxidant, anti-inflammatory and cholinesterase inhibitory activities

UPLC-PDA-MS/MS Profiling and Healing Activity of Polyphenol-Rich Fraction of Alhagi maurorum against Oral Ulcer in Rats

Camelthorn, Alhagi maurorum Boiss, family Fabaceae has long been used in African folk medicine owing to its richness in pharmacologically active metabolites. The crude extract (CEAM), ethyl acetate fraction (EFAM) and n-butanol (BFAM) fraction of A. maurorum aerial parts were investigated for their total polyphenols and oral antiulcer activity using in-vitro and in-vivo models. The major phenolic compound was isolated from the polyphenol-rich EFAM fraction and identified by conventional and spectroscopic methods of analysis as isorhamnetin-3-O-rutinoside. Furthermore, standardization of EAFM using UPLC-PDA-UV quantified isorhamnetin-3-O-rutinoside as 262.91 0.57 g/mg of the fraction. Analysis of EFAM using UPLC-PDA-MS/MS revealed tentative identification of 25 polyphenolic compounds. EFAM exhibited the most potent free radical scavenging activity against DPPH, with an IC50 (27.73 ± 1.85 µg/mL) and an FRAP value of (176.60 ± 5.21 μM Trolox equivalent (TE)/mg fraction) in comparison with CEAM and BFAM. Acetic acid-induced oral ulcers in a rat model were used to evaluate the healing properties of A. maurorum aerial parts. EFAM significantly decreased tumor necrosis factor-alpha (TNF-α) and interleukin-2 (IL-2) by 36.4% and 50.8%, respectively, in the ulcer tissues while, CEAM and BFAM exhibited lower activity at the same dose. In addition, EFAM led to a significant (p < 0.0001) rise in the expression of proliferating cell nuclear antigen (PCNA), a cell proliferation marker. A. maurorum exhibited a potent healing effect in acetic acid-induced oral ulcers in rats by mitigating the release of pro-inflammatory cytokines and improving PCNA expression

Effects of 3-NPA and/or genistein on striatal, cortical and hippocampal AChE activity of ovariectomized rats.

<p>3-NPA was administered i.p. (20 mg/kg) for 4 consecutive days. 17β-estradiol (2.5 mg/kg body weight, s.c) and genistein (10 and 20 mg/kg body weight, i.p) were administered for 8 days, beginning 4 days before and continued for 4 days one hour before 3-NPA injections. Data are presented as means ±S.E.M. (n = 6). ^x statistically significant compared to sham group at P< 0.05, * statistically significant compared to control group at P< 0.05, ^# statistically significant compared to 3-NPA-treated group at P< 0.05 (one-way ANOVA followed by Tukey test).</p

Effects of 3-NPA and/or genistein on striatal, cortical and hippocampal AChE activity of ovariectomized rats.

<p>3-NPA was administered i.p. (20 mg/kg) for 4 consecutive days. 17β-estradiol (2.5 mg/kg body weight, s.c) and genistein (10 and 20 mg/kg body weight, i.p) were administered for 8 days, beginning 4 days before and continued for 4 days one hour before 3-NPA injections. Data are presented as means ±S.E.M. (n = 6). ^x statistically significant compared to sham group at P< 0.05, * statistically significant compared to control group at P< 0.05, ^# statistically significant compared to 3-NPA-treated group at P< 0.05 (one-way ANOVA followed by Tukey test).</p

Effects of 3-NPA and/or genistein on locomotor activity in ovariectomized rats.

<p>3-NPA was administered i.p. (20 mg/kg) for 4 consecutive days. 17β-estradiol (2.5 mg/kg body weight, s.c) and genistein (5, 10 and 20 mg/kg body weight, i.p) were administered for 8 days, beginning 4 days before and continued for 4 days one hour before 3-NPA injections. Data are presented as means ±S.E.M. (n = 8). ^x statistically significant compared to sham group at P< 0.05, * statistically significant compared to control group at P< 0.05, ^# statistically significant compared to 3-NPA-treated group at P< 0.05 (two-way ANOVA followed by Bonferroni test).</p

H & E staining of the cortices of rats belonging to the sham group (A), control group (B), 3-NPA-treated group (C), 17β-estradiol- and 3-NPA-treated group (D), genistein (5 mg/kg)- and 3-NPA-treated group (E), genistein (10 mg/kg)- and 3-NPA-treated group (F), genistein (20 mg/kg)- and 3-NPA-treated group (G) and genistein alone-treated group (H): A, B, D and H showed no histological alterations, C showed severe hemorrhage (h), E showed focal gliosis (g) F showed focal gliosis (g) and G showed congested blood vessel (C).

<p>H & E staining of the cortices of rats belonging to the sham group (A), control group (B), 3-NPA-treated group (C), 17β-estradiol- and 3-NPA-treated group (D), genistein (5 mg/kg)- and 3-NPA-treated group (E), genistein (10 mg/kg)- and 3-NPA-treated group (F), genistein (20 mg/kg)- and 3-NPA-treated group (G) and genistein alone-treated group (H): A, B, D and H showed no histological alterations, C showed severe hemorrhage (h), E showed focal gliosis (g) F showed focal gliosis (g) and G showed congested blood vessel (C).</p

Immunohistochemical staining of cortical COX-2-positive cells immunized with goat-anti-rabbit antibodies of the sham group (A), control group (B), 3-NPA-treated group (C), 17β-estradiol- and 3-NPA-treated group (D), genistein (10 mg/kg)- and 3-NPA-treated group (E), genistein (20 mg/kg)- and 3-NPA-treated group (F) and genistein alone-treated group (G).

<p>Immunohistochemical staining of cortical COX-2-positive cells immunized with goat-anti-rabbit antibodies of the sham group (A), control group (B), 3-NPA-treated group (C), 17β-estradiol- and 3-NPA-treated group (D), genistein (10 mg/kg)- and 3-NPA-treated group (E), genistein (20 mg/kg)- and 3-NPA-treated group (F) and genistein alone-treated group (G).</p

Effects of 3-NPA and/or genistein on step through passive avoidance in ovariectomized rats.

<p>3-NPA was administered i.p. (20 mg/kg) for 4 consecutive days. 17β-estradiol (2.5 mg/kg body weight, s.c) and genistein (5, 10 and 20 mg/kg body weight, i.p) were administered for 8 days, beginning 4 days before and continued for 4 days one hour before 3-NPA injections. Training was performed on day 5. First and second retention latencies were assessed at days 6 (A) and 8 (B). Data are presented as medians and quartiles (n = 8). ^x statistically significant compared to sham group at P< 0.05, * statistically significant compared to control group at P< 0.05, ^# statistically significant compared to 3-NPA-treated group at P< 0.05 (Kruskal-Wallis nonparameteric test followed by Dunn’s test).</p

H & E staining of the hippocampi of rats belonging to the sham group (A), control group (B), 3-NPA-treated group (C), 17β-estradiol- and 3-NPA-treated group (D), genistein (5 mg/kg)- and 3-NPA-treated group (E), genistein (10 mg/kg)- and 3-NPA-treated group (F), genistein (20 mg/kg)- and 3-NPA-treated group (G) and genistein alone-treated group (H): A, B, D, F, G and H showed no histological alterations, C showed severe neurodegeneration (d) and hemorrhage (h) and E showed some degenerated hippocampal cells (d).

<p>H & E staining of the hippocampi of rats belonging to the sham group (A), control group (B), 3-NPA-treated group (C), 17β-estradiol- and 3-NPA-treated group (D), genistein (5 mg/kg)- and 3-NPA-treated group (E), genistein (10 mg/kg)- and 3-NPA-treated group (F), genistein (20 mg/kg)- and 3-NPA-treated group (G) and genistein alone-treated group (H): A, B, D, F, G and H showed no histological alterations, C showed severe neurodegeneration (d) and hemorrhage (h) and E showed some degenerated hippocampal cells (d).</p