Nuclear factor erythrogen-2 associated factor 2 (Nrf2) signaling is an essential molecular pathway for the anti-aging effect of whey protein in the prefrontal cortex of aging rat model (Histological and Biochemical Study).

Aging is a highly complicated natural process. Brain aging is associated with remarkable neurodegenerative changes and oxidative damage. Whey protein (WP) has been mentioned to have an antioxidant property. Nuclear factor erythrogen-2 associated factor 2 (Nrf2) signaling pathway is an antioxidant defense system. Nrf2 activity declines with age so, its activation could be a promising therapeutic strategy for aging. This study aimed to explore the anti-aging role of WP against D-galactose (D-gal) induced age-related degenerative changes and oxidative damage in the prefrontal cortex (PFC) and investigate its underlying mechanisms. Forty adult male rats were divided into 4 groups; control, WP group received WP (28.77 mg/kg/day) by gastric tube on the 4th experimental week; D-gal (model group) received D-gal (300 mg/kg/day) intraperitoneally for 8 weeks and D-gal +WP group received WP on the 4th week of D-gal treatment. Specimens from PFC were obtained for biochemical, histological, immunohistochemical and western blot analysis. WP treatment in D-gal +WP group reduced lipid peroxidation, enhanced antioxidant enzyme activities, decreased advanced glycation end products level and improved the histological and ultrastructural alterations. Moreover, the number of neurons expressed the senescence marker; p21 and percentage area of the astrocytic marker; glial fibrillary acidic protein were significantly reduced. WP also enhanced Nrf2 pathway and its downstream targets; heme oxygenase-1 and NADPH quinone oxidoreductase 1. In conclusion WP alleviates the D-gal-induced PFC aging through activating Nrf2 pathway, reducing cell senescence and gliosis. So, it may be a potential therapeutic target to retard the aging process.

Quercetin mitigates monosodium glutamate-induced excitotoxicity of the spinal cord motoneurons in aged rats via p38 MAPK inhibition.

Various studies reported the possibility of deterioration of blood-brain barrier (BBB) integrity owing to the aging process. The current work was performed to investigate the ability of Monosodium glutamate (MSG) to cross BBB in aged rats, the damage affecting the anterior horn cells of the spinal cord due to excitotoxicity, and the mechanisms by which quercetin (Que) administration might suppress such damage. Forty male rats aged 18 months were assigned equally to 4 groups: control group, Que group (received Que, 20 mg/kg/d intraperitonealy for 10 days), MSG group (received MSG, 4.0 g/kg/d subcutaneously for 10 days), MSG + Que group (received both Que and MSG as done in the Que and MSG groups respectively). Cervical spinal cord specimens were obtained and prepared for routine histological study, immunohistochemical staining by caspase-3 and glial fibrillary acidic protein (GFAP), assessment of oxidative stress, measurement of cytokines, assessment of caspase-3 activity and GFAP levels as well as for western blotting of phosphorylated activating transcription factor 2 (ATF2pp) as an indicator for the activity of p38 mitogen-activated protein kinase (MAPK). The MSG group revealed variable degenerative and apoptotic changes in the motoneurons and neuroglia, a marked rise in the cytoplasmic caspase-3 expression in motoneurons and a significant reduction (p < 0.001) in the astrocyte surface area percentage. In addition, the spinal cord tissue exhibited a significant elevation (p < 0.001) in the levels of malondialdehyde (MDA), IL-1, IL-6, TNFα, INFÉ£, caspase-3 activity and ATF2 pp expression as well as a significant reduction (p < 0.001) in SOD, IL-10 and GFAP levels compared with the control group. On combining Que with MSG, most of the degenerative changes were reversed and all the impaired parameters were nearly normalized except for IL-6 and GFAP levels which were still significantly (p < 0.05) different from those of the control group. Our study suggests that MSG can break through the BBB of the aged rats and induce excitotoxicity dependent changes in spinal cord motoneurons. Most of these changes were reversed by Que probably via targeting the p38 MAPK-ATF2 pathway, antagonizing oxidative stress, anti-inflammatory effect, and promoting GFAP expression.

The Ameliorating Effect of Sodium Selenite on the Histological Changes and Expression of Caspase-3 in the Testis of Monosodium Glutamate-Treated Rats: Light and Electron Microscopic Study.

Monosodium glutamate (MSG) is a commonly used flavor enhancer that may contribute to male infertility. Sodium selenite is inorganic chemical form of selenium (Se). Se is best known as an antioxidant. This study was designed to investigate the possible ameliorating role of sodium selenite against MSG-induced testicular toxicity and histological changes. Forty male albino rats were allocated into four groups. Control group received distilled water, SE group received sodium selenite (0.25 mg/kg/day) dissolved in distilled water orally, MSG group received MSG (6 mg/g/day) dissolved in distilled water orally, and MSG + SE group received both MSG and sodium selenite for 45 days. Testicular samples were prepared for biochemical, light, and electron microscopic studies. Immunohistochemical staining for caspase-3 was done. MSG group demonstrated a significant increase in malondialdehyde level, marked damage of seminiferous tubules with a significant reduction in diameter and height of the lining epithelium. Spermatogenic cells showed disorganization, dark nuclei, reduction in number, and maturation arrest. Vacuolations of interstitial tissue and Leydig cells were also observed. Percent area of fibrosis and caspase-3 immunoexpression was significantly increased. Ultrastructurally, irregular tubular basement membrane and damaged germ cells were found. The spermatogenic, Sertoli, and Leydig cells showed irregular shrunken nuclei, cytoplasmic vacuolations, and swollen mitochondria. MSG + SE group showed much better histological and ultrastructural picture and improvement of the measured biochemical and morphometric parameters. Percent area of caspase-3 immunoexpression was significantly decreased. In conclusion, sodium selenite ameliorated the testicular damaging effect of MSG through reduction of oxidative stress and apoptosis.

An immunohistochemical and ultrastructural analysis of the retina in tadalafil (Cialis) treated rats.

Tadalafil (Cialis) is one of the most commonly used phosphodiesterase type5 (PDE5) inhibitors. This work aimed to analyze the histological and ultrastructural changes provoked by chronic tadalafil administration in the rat retina, correlate between such changes and PDE5 immunoexpression and to evaluate the possible reversibility of these changes. Thirty Sprague Dawley male rats were randomly distributed into 3 groups. Control group; given 1 ml distilled water daily for 6 weeks. Tadalafil group; given tadalafil in a daily dose of 2.6 mg/kg for 6 weeks. Withdrawal group; given tadalafil 2.6 mg/kg daily for 6 week followed by a withdrawal period of 4 weeks. Retinal specimens were prepared for histological, ultrastructural and immunohistochemical study using anti-PDE5 and anti-Bcl-2 antibodies. Morphometric and statistical studies were performed. Tadalafil group displayed a significant reduction in retinal thickness, diminished cell population of outer and inner nuclear layers, dilated blood capillaries and a significant decline in the number of ganglion cells. Significant reductions of both PDE5 and Bcl-2 immunoexpression were observed. At the ultrastructural level, the photoreceptors showed spacing of outer segments and disorganized membranous discs. Retinal neurons showed ultrastructural degenerative changes in the form of shrunken irregular nuclei, dilated rER, and disrupted mitochondria. Withdrawal group revealed preservation of histological structure and partial restoration of retinal thickness, retinal cells ultrastructure, and PDE5 and Bcl-2 immunoexpressions. In conclusion, chronic use of tadalafil could induce reversible apoptotic and degenerative changes in retinal neurons due to its inhibitory effect on PDE5 expression which affects the metabolism and viability of retinal cells.

Deletion of Thioredoxin-interacting protein ameliorates high fat diet-induced non-alcoholic steatohepatitis through modulation of Toll-like receptor 2-NLRP3-inflammasome axis: Histological and immunohistochemical study.

Endemic prevalence of obesity is associated with alarming increases in non-alcoholic steatohepatitis (NASH) with limited available therapeutics. Toll-like receptor2 (TLR2) and Nod-like receptor protein 3 (NLRP3) Inflammasome are implicated in hepatic steatosis, inflammation and fibrosis; the histological landmark stages of NASH. TXNIP, a member of α-arrestin family activates NLRP3 in response to various danger stimuli. The aim of current work was to investigate the effect of TXNIP genetic deletion on histological manifestations of high fat diet-induced steatohepatitis and activation of TLR2-NLRP3-inflammasome axis. Wild-type mice (WT) and TXNIP knock out (TKO) littermates were randomized to normal diet (WT-ND and TKO-ND) or high fat diet (HFD, 60% fat) (WT-HFD and TKO-HFD). After 8-weeks, liver samples from all groups were evaluated by histological, immunohistochemical and western blot analysis. HFD resulted in significant induction of micro and macrovesicular hepatic steatosis, that was associated with increased inflammatory immune cell infiltration in WT-HFD compared with WT-ND and TKO-ND controls, but not in TKO-HFD group. In parallel, WT-HFD group showed significant fibrosis and α-SMA expression; a marker of pro-fibrotic stellate-cell activation, in areas surrounding the central vein and portal circulation, versus all other groups. Western blot revealed increased activation of TLR2-NLRP3 inflammasome pathway and downstream IL-1ß and TNFα in WT-HFD group, but not in TKO-HFD group. IL-1ß expression coincided within the same areas of steatosis, inflammatory cell infiltration, collagen deposition and α-SMA expression in WT-HFD mice, that was significantly reduced in TKO-HFD mice. In conclusion, TXNIP deletion ameliorates the HFD-induced steatosis, inflammatory and fibrotic response via modulation of TLR2-NLRP3 inflammasome axis. Targeting TXNIP-TLR2-NLRP3 pathway may provide potential therapeutic modalities for NASH treatment.

Histological and immunohistochemical study of cardiac telocytes in a rat model of isoproterenol-induced myocardial infarction with a reference to the effect of grape seed extract.

INTRODUCTION: Cardiac telocytes (TCs) represent a unique type of cells that make a supportive network for stem cells that contribute in cardiac renewal, but their role during myocardial infarction (MI) is not clear. Grape seed extract (GSE) is a powerful natural antioxidant. AIM OF THE WORK: Quantitative study of cardiac TCs in a rat model of Isoproterenol (ISO)-induced MI, and to evaluate the effect of GSE on TCs and MI progression. MATERIALS AND METHODS: Seventy adult male albino rats were assigned into 4 groups; group I; control rats, group II received GSE (100mg/kg/day) dissolved in distilled water orally, group III received 2 intra-peritoneal injections of 85mg/kg ISO dissolved in saline on 14th and 15th day to induce MI, and group IV received GSE and ISO. Myocardium was obtained 1 and 14days after ISO i.e. on day 16 and day 30 respectively. Tissue was prepared for histological and immunohistochemical study of CD117 and CD34 as two markers for TCs. CD34 was used also as a marker for angiogenesis. RESULTS: Group III showed focal areas of myocardial infarction 1day and 14days after ISO. Degenerated cardiomyocytes showed loss of striation and hypereosinophilic vacuolated cytoplasm with condensed nuclei. Mononuclear cell infiltration and a significantly increased percentage area of fibrosis 14days after ISO were observed. CD117 and CD34 positive TCs were hardly detected 1day after ISO. Their number slightly increased 14days after ISO with insignificant difference to control. There was also a significant increase in the number of CD34 positive blood vessels 14days after ISO. Group IV showed much better histological picture with a significant decrease in the percentage area of fibrosis and a significant increase in the number of CD117 and CD34 positive TCs and the number of CD34 positive blood vessels as compared to group III. CONCLUSION: Telocytes were significantly decreased in MI. GSE reduced ISO-induced histological changes and increased the number of TCs that improved angiogenesis.

The possible protective role of pumpkin seed oil in an animal model of acid aspiration pneumonia: Light and electron microscopic study.

Aspiration pneumonitis is a common problem occurring in many clinical disorders. Pumpkin seed oil (PO) is a rich source of antioxidants. This work aimed to assess the effect of PO on the lung histopathological changes induced by acid aspiration. Forty male albino rats assigned to four groups were used. Rats of control group were instilled intratracheally with normal saline 2mL/kg. HCL group instilled with 2mL/kg of HCL 0.1N, pH 1.25. PO group received pumpkin seed oil (PO) orally (∼1375mg/kgbw/day) for 7days. HCL+PO group instilled with 2mL/kg of HCL 0.1N, pH 1.25 and received PO at the same dose of PO group. Lung tissue samples were processed for light, electron microscopic and immunohistochemical study using anti inducible NO synthase (iNOS). The lung of HCL group demonstrated thickened interalveolar septa, inflammatory cell infiltration and significant increase in the area percent of collagenous fibers and immune expression of iNOS. Ultra structurally, disrupted alveolocapillay membrane, degenerated type II pneumocytes and plentiful alveolar macrophages were evident. PO administration partially attenuated these histological and ultra structural alterations and reduced iNOS immune-expression in lung tissue. In conclusion, PO has a protective effect against HCL aspiration lung injury most probably through its antioxidant activity.