Thymus daenensis extract prevents scopolamine­induced memory impairment through declining oxidative stress in rats.

Memory and cognitive impairment induced by oxidative stress are among the main hallmarks of Alzheimer's disease's (AD) pathology. The present study aimed to investigate the potential neuroprotective effects of Thymus daenensis (T. daenensis) extract against scopolamine­induced memory impairment and oxidative stress in rats. T. daenensis, widely distributed in Iran and Europe, is known to be a rich source of natural antioxidants and has been traditionally used for various medical purposes. The present study investigated the post­treatment effects of T. daenensis on learning and memory functions, antioxidant cellular defense, and oxidative stress using the scopolamine rat model of AD. The experiments were performed by intraperitoneal injection of scopolamine for 10 consecutive days in Wistar male rats (180-220 g). Additionally, the animals received T. daenensis extract (50­200 mg/kg) by gavage for 14 consecutive days after induction of memory impairment. The animals were divided into 8 groups, namely: control, 200 mg/kg of T. daenensis extract (D200), donepezil (DON), scopolamine (ALZ), ALZ animals treated with different doses of the extract (ALZ+D50 or 100 or 200 mg/kg) and ALZ animals treated with (ALZ+DON). The animals were then subjected to the Morris water maze (MWM) paradigm as a standard criterion for memory function assessment, and after extracting the brain tissues, the related biochemical oxidative stress parameters were determined in the brain. Our results indicated that T. daenensis extract significantly improved animals' performance in the MWM while significantly reducing oxidative stress and antioxidant imbalance. Furthermore, the extract did not show hepatotoxic effects on treated animals. In addition, the extract treatment significantly decreased both cellular malondialdehyde (MDA) and protein carbonyl (PCO) content while conversely increasing the total reduced glutathione (GSH) content and also the levels of total and endogenous antioxidants in the ferric reducing antioxidant power (FRAP) assay. It seems that the administration of T. daenensis significantly improved both cellular biochemical aspects and memory performance in animal models. Conclusively, it could be beneficial for scopolamine­induced neurotoxicity.

Protective effects of Vinca herbaceous extract against scopolamine-induced behavioral disturbances and brain oxidative stress in rats.

Ethnopharmacological relevance: Alzheimer's disease (AD) as the most common type of dementia, is affecting the life of many senior individuals around the world. Vinca herbacea Waldst. & Kit. (V. herbacea) as a middle east originated plant demonstrated antioxidant and antitumor effects. This plant traditionally used to treat diabetes and hypertension, but its mechanism remains unclear. Aim of the study: In the present study, post-treatment effects of V. herbacea on learning and memory functions, antioxidant cellular defense and oxidative stress were investigated using the scopolamine rat model of AD. Materials and methods: Wistar male rats (170-190 g) were administered Scopolamine, an anti-muscarinic drug, (2 mg/kg) for 10 days followed by V. herbacea extract (200, 300 and 400 mg/kg) and/or donepezil (DON; 1 mg/kg, which were administered before behavioral studies for 10 consecutive days. All the rats were then subjected to Morris water maze (MWM) task. Biochemical parameters of oxidative stress were quantified using the whole brain. Results: Our data showed significant decrease performance in target quadrant in water maze task following administration of scopolamine (SCOP). Also, V. herbacea and DON, did not induce any neurotoxicity and hepatotoxic effects at the highest utilized doses in healthy rats. Treatment with V. herbacea extract (200&400 mg/kg) and DON improved memory performance significantly in comparison with AD rats. In addition, V. herbacea extract in AD rats exhibited a decrease in malondialdehyde (MDA) and protein carbonyl (PCO) levels and an increase in total antioxidant capacity (FRAP) and glutathione (GSH) amounts in brain and liver. Conclusion: It seems that cholinergic deficits and oxidative stress are consistently associated with Alzheimer's disease (AD). The richness of V. herbacea in case of indole alkaloids and flavonoids confirms the potentials of this herb in management of oxidative stress, resorting synaptic acetylcholine level and improving cellular antioxidant resources.

Investigation of therapeutic effect of curcumin α and ß glucoside anomers against Alzheimer's disease by the nose to brain drug delivery.

Alzheimer's disease (AD) is one of the greatest geriatric medicinal challenges of our century and is the main disease leading to dementia. Despite extensive scientific research advances, available disease-modifying treatment strategies remained limited; thus, increasing demand for new drugs. In recent years, medicinal plants attracted attention due to their potential role in dementia. In the present study, α and ß anomers of curcumin glucosides (CGs) were synthesized and evaluated for Alzheimer's treatment. CGs were synthesized by fusion reaction as a novel and easy method with more advantages (high yield, short reaction time, and low chemicals), and the products were characterized using HNMR. Wistar male rats were used to administer different treatments. They divided into control, sham, Alzheimer, and test groups (Alzheimer + α anomer and Alzheimer + ß anomer). Animals received normal saline, Scopolamine (1 mg/kg), high dose anomers, scopolamine, and two doses (12.5 and 25 mg/kg) of anomers, respectively, for 10 days. Then the Morris Water Maze (MWM) test was performed on all animals. Finally, the animals' brains were extracted and homogenized for glutathione, acetylcholine esterase activity, protein carbonyl, and lipid peroxide level detection. The escape latency and the distance towards the hidden platform in Morris water maze in the Alzheimer group were significantly higher than both the control and test groups. Besides, there were no significant differences between sham and control groups in all tests. Both anomers led to a significant increase in glutathione, and acetylcholine levels while they caused a decrease in lipid peroxidation and protein carbonyl levels in brain tissue. It seems that intranasal administration of both anomers positively influenced maze learning in scopolamine receiving subjects. Although both anomers resulted in similar biochemistry tests, a higher dose of ß anomer indicated better results than α anomer not only in behavioral tests but also in biochemical tests.

The protective effects of Ziziphora tenuior L. against chlorpyrifos induced toxicity: Involvement of inflammatory and cell death signaling pathway.

ETHNOPHARMACOLOGICAL RELEVANCE: Ziziphora tenuior L. is used as a medicinal plant in treatment of various diseases such as gastric disorders, stomach ache, dysentery, uterus infection, gut inflammation and menstruation. AIM OF THE STUDY: In the present study, the protective effects of Ziziphora tenuior extract against chlorpyrifos (CPF), the most commonly or popularly used insecticide in Asia and Africa were investigated in liver and lung tissues with emphasis in apoptotic and inflammatory pathways in rat model. MATERIALS AND METHODS: The experiments were performed by gavage of male rats for 8 weeks. The extract of Z. tenuior was administrated at three different doses (40, 80, 160 mg/kg). 6.75 mg/kg CPF was administrated as the maximum tolerable dose based on our previous study. RESULTS: Our data indicated that CPF can increase the expression of some inflammatory genes (IL-6, TLR-2, IL-1ß, TNF-α, and NLPR3) and apoptosis genes (Caspase 3, Caspase 9, Caspase 8 and Bax). On the other hand, it can down regulate Bcl-2 gene expression. Post-treatment of Z. tenuior extract in CPF- treated rats showed significant decrease in apoptotic and inflammatory gene expression in the liver and lung due to its anti-apoptotic effects which confirmed by Bcl-2 gene overexpression. CONCLUSION: The present study suggested that Z. tenuior extract, as a traditional treatment can be able to moderate CPF toxicity via significant effect on inflammatory and apoptotic cell death signaling pathway. Also, based on our preliminary data, it is suggested that Z. tenuior extract can prevent the adverse effects of CPF in liver and lung tissues.

Grape Seed Oil as a Natural Therapy in Male Rats with Alzheimer's Diseases.

Purpose: Alzheimer's disease (AD) is a chronic neurodegenerative disorder, with an increasing prevalence rate, mostly related to cholinergic system. According to the difficulties and complications in management of AD, this study was carried out to evaluate the efficacy of grape seed oil (GSO) on scopolamine (Scop) induced Alzheimer's in male rats. Methods: 64 healthy male Wistar rats received different treatments such as: normal saline (NS), donepezil (Don), Scop and GSO, according to the previously designed protocol. Morris (MWM) was applied for spatial memory tests. Right after the behavioral tests, the brains were removed and the hippocampus was separated for evaluation of acetylcholine levels as well as cell death and neuro inflammation. Results: The results of the test day indicated that the mean Q2 time was increased in both GSO test groups (P <0.05) and Don treated group (P <0.001).The spectrophotometric findings affirm that both GSO co-treatment and post-treatment were effective in augmenting brain acetylcholine levels (P <0.01 and P <0.05 respectively). The microscopic findings of H&E dyed tissues confirmed the above mentioned results for different treatments except for GSO post treatment, in which the viability of cells were very low. Conclusion: The results implied that supplementation of rats with GSO caused a significant augmentation in spatial memory performance as well as acetylcholine levels and cell viability in the presence of Scop. This effect was comparable to that of Don especially when GSO was used as co-treatment.

Selegiline (L-Deprenyl) Mitigated Oxidative Stress, Cognitive Abnormalities, and Histopathological Change in Rats: Alternative Therapy in Transient Global Ischemia.

Selegiline (L-deprenyl) is the major drug which is used in the treatment of Parkinson's disease because of its neurotrophic and antiapoptotic properties. Previous studies suggested that low dose of L-methamphetamine (L-METH) caused lower mortality rate in patients with severe traumatic brain injury. As L-methamphetamine is one of the metabolites of selegiline, the present study aims to examine whether L-deprenyl can improve cognitive, biochemical, and histopathological injury in animal model of transient global ischemia. The animals were randomized in ten groups orally gavaged three times a week for 28 days. Then, novel object recognition (NOR) was conducted to assess their behavioral abnormality. After scarification of the rats, their brains were divided into two sections to measure oxidative stress parameters and perform pathological evaluations in rats. Our data revealed the involvement of oxidative stress, behavioral despair, and pathological data in transient global ischemia rats. Significant recovery in cognitive behavior, oxidative stress biomarker, and number of dead cell in histopathological assay was observed in rats treated with 1,2 and 4 mg/kg of selegiline. So, selegiline appears to be useful in alternative therapy of transient global ischemia.

Coenzyme Q10 Alleviated Behavioral Dysfunction and Bioenergetic Function in an Animal Model of Depression.

Coenzyme Q10 (CoQ10) is a natural compound, is involved in the mitochondrial electron transfer chain (ETC) and plays an important pattern in adenosine triphosphate (ATP) production. Amelioration of ATP is related to abnormalities in cognitive function and psychiatric diseases. Previous studies have shown that depression is accompanied by the induction of inflammatory and oxidative stress pathways and amelioration of antioxidant status. In a recent study, we investigated the beneficial effects of CoQ10 on behavioral dysfunction and CoQ10 level in the rat brain. Therefore, intracerebroventricular (ICV) infusion of a single dose of streptozotocin (STZ, 0.2 mg/mouse) was used in adult male mice to induce depression. The behavioral data revealed a significant difference between the depression and control groups regarding the forced swim test (FST) and splash test results at 24 h following STZ treatment. Also, the validated and accurate high-performance liquid chromatography (HPLC) technique showed decreased CoQ10 level in the brain samples of the STZ group, compared to the controls. Our findings revealed that behavioral abnormalities due to STZ target mitochondria and affect energy metabolism and hemostasis, resulting in the initiation of oxidative damage in the brain. Besides, 4-week administration of CoQ10 could reverse the depressive like behavior and bioenergetic effects of STZ in the treated groups.

Adjuvant potential of selegiline in treating acute toxicity of aluminium phosphide in rats.

Aluminium phosphide (AlP) is a highly toxic substance with a high mortality rate and no effective antidote. Once exposed to the moisture and acidic conditions of the stomach, AlP releases toxic phosphine (PH3 ) gas, which results in severe toxicity in poisoned subjects. Selegiline is a monoamine oxidase inhibitor with antioxidant and anti-apoptotic properties, which is mostly prescribed for the treatment of mood disorders and Parkinson's disease. Since AlP has detrimental effects on cardiac physiology and mitochondrial function, we tested the protective effects of acute selegiline treatment on cardiac mitochondrial function, redox status and electrocardiographic parameters in rats after AlP poisoning. To do this, AlP was given to rats by gavage to induce toxicity. Selegiline was injected intraperitoneally in the treatment groups 1 hour after AlP poisoning. Selegiline treatment after AlP intoxication was not associated with a significant difference in the mortality rate of animals. However, selegiline reduced oxidative stress (decreased the reactive oxygen species and malondialdehyde) and increased glutathione in the cardiac tissue of rats exposed to AlP. Further, the mitochondrial membrane potential (ΔΨm) collapse reversed after treatment with selegiline. Selegiline also improved the electrocardiographic (ECG) parameters and enhanced heart rate. The histopathological evaluation revealed that selegiline eliminated the inflammation and injuries induced by AlP in the stomach and duodenum, as well as cardiac tissue. In conclusion, selegiline treatment can ameliorate the AlP-induced cardiac and gastrointestinal injuries in rats via boosting redox status and mitochondrial function with no significant effect on survival. We suggest that using selegiline, apart from other clinical treatments, may improve the quality of treatment process for AlP toxicity.

Protective effects of Ziziphora tenuior extract against chlorpyrifos induced liver and lung toxicity in rat: Mechanistic approaches in subchronic study.

Chlorpyrifos (CPF) is one of the most widely used organophosphorus, which has spurred renewed interest. This study was conducted to investigate the protective effect of ziziphora tenuior extract against CPF-induced liver and lung toxicity. This study conducted 8-week rat sub-chronic toxicity study and then the effect of ziziphora tenuior extract in 3 different doses (40, 80, 160 mg/kg) was determined. We administrated maximum tolerated dose of CPF (6.75 mg/kg) by gavage for 8 weeks (5 times in week) to male rats. Rats were sacrificed 24 h after last dose and the biochemical analysis, which confirms involvement of oxidative stress in the pathogenesis of CPF toxicity in liver including increased in lipid peroxidation, protein carbonyl content, and ROS formation, glutathione depletion, decreased of antioxidant effect via frap oxidation and cytochrome c expulsion. In addition, pathological lesions confirm the dysfunction of the organs (liver and lung). In addition, using of ziziphora extract as an antioxidant is resulted in amelioration of oxidative stress marker in liver and lung damage. In conclusion, the current study revealed that CPF toxicity is related to oxidative stress and induction of cell death signaling and cotreatment with ziziphora extract is recommended in the routine therapy for the protection against CPF induced liver and lung tissue damage.

Mitochondria as a Target for the Cardioprotective Effects of Cydonia oblonga Mill. and Ficus carica L. in Doxorubicin-Induced Cardiotoxicity.

Quince (Cydonia oblonga Mill.) and fig (Ficus carica L.) exhibit a broad spectrum of pharmacological activities. Regarding the cardiotoxic effect of doxorubicin (DOX) is mediated mainly through mitochondrial oxidative stress and dysfunction; the present study evaluated the cardioprotective effects of the aqueous extracts of Cydonia oblonga Mill. fruit (ACO) and Ficus carica L. fruit (AFC) against DOX-induced cardiotoxicity. Cardiomyocytes toxicity was induced in male Sprague Dawley rats by intraperitoneal (ip) injections of 2.5 mg/kg DOX 3 times per week for a period of 2 weeks. After heart failure was induced in the rats, the animals were decapitated and their hearts were immediately removed. Then, the cardiac mitochondria were isolated by differential ultracentrifugation, and the protective effects of each particular extract on mitochondrial oxidative stress and dysfunction were determined. ACO and AFC ameliorated mitochondrial dysfunction in the isolated mitochondria and prevented mitochondrial reactive oxygen species formation, membrane lipid peroxidation, mitochondrial swelling, mitochondrial membrane potential collapse (%ΔΨm), and cytochrome c release. Also, the extracts significantly increased reduced glutathione levels and succinate dehydrogenase activity. These results indicated that ACO and AFC have beneficial effects against DOX cardiotoxicity which mediated by attenuating mitochondrial dysfunction. Therefore, it can be suggested that quince and fig may increase the therapeutic index of DOX.

Protective effects of Sesamum indicum extract against oxidative stress induced by vanadium on isolated rat hepatocytes.

Vanadium toxicity is a challenging problem to human and animal health with no entirely understanding cytotoxic mechanisms. Previous studies in vanadium toxicity showed involvement of oxidative stress in isolated liver hepatocytes and mitochondria via increasing of ROS formation, release of cytochrome c and ATP depletion after incubation with different concentrations (25-200 µM). Therefore, we aimed to investigate the protective effects of Sesamum indicum seed extract (100-300 µg/mL) against oxidative stress induced by vanadium on isolated rat hepatocytes. Our results showed that quite similar to Alpha-tocopherol (100 µM), different concentrations of extract (100-300 µg/mL) protected the isolated hepatocyte against all oxidative stress/cytotoxicity markers induced by vanadium in including cell lysis, ROS generation, mitochondrial membrane potential decrease and lysosomal membrane damage. Besides, vanadium induced mitochondrial/lysosomal toxic interaction and vanadium reductive activation mediated by glutathione in vanadium toxicity was significantly (P < 0.05) ameliorated by Sesamum indicum extracts. These findings suggested a hepato-protective role for extracts against liver injury resulted from vanadium toxicity. © 2015 Wiley Periodicals, Inc. Environ Toxicol 31: 979-985, 2016.

Depleted uranium induces disruption of energy homeostasis and oxidative stress in isolated rat brain mitochondria.

Depleted uranium (DU) is emerging as an environmental pollutant primarily due to its military applications. Gulf War veterans with embedded DU showed cognitive disorders that suggest that the central nervous system is a target of DU. Recent evidence has suggested that DU could induce oxidative stress and mitochondrial dysfunction in brain tissue. However, the underlying mechanisms of DU toxicity in brain mitochondria are not yet well understood. Brain mitochondria were obtained using differential centrifugation and were incubated with different concentrations (50, 100 and 200 µM) of uranyl acetate (UA) as a soluble salt of U(238) for 1 h. In this research, mitochondrial ROS production, collapse of mitochondrial membrane potential and mitochondrial swelling were examined by flow cytometry following the addition of UA. Meanwhile, mitochondrial sources of ROS formation were determined using specific substrates and inhibitors. Complex II and IV activity and also the extent of lipid peroxidation and glutathione (GSH) oxidation were detected via spectroscopy. Furthermore, we investigated the concentration of ATP and ATP/ADP ratio using luciferase enzyme and cytochrome c release from mitochondria which was detected by ELISA kit. UA caused concentration-dependent elevation of succinate-linked mitochondrial ROS production, lipid peroxidation, GSH oxidation and inhibition of mitochondrial complex II. UA also induced mitochondrial permeability transition, ATP production decrease and increase in cytochrome c release. Pre-treatment with antioxidants significantly inhibited all the above mentioned toxic effects of UA. This study suggests that mitochondrial oxidative stress and impairment of oxidative phosphorylation in brain mitochondria may play a key role in DU neurotoxicity as reported in Gulf War Syndrome.

Toxicity of depleted uranium on isolated rat kidney mitochondria.

BACKGROUND: Kidney is known as the most sensitive target organ for depleted uranium (DU) toxicity in comparison to other organs. Although the oxidative stress and mitochondrial damage induced by DU has been well investigated, the precise mechanism of DU-induced nephrotoxicity has not been thoroughly recognized yet. METHODS: Kidney mitochondria were obtained using differential centrifugation from Wistar rats and mitochondrial toxicity endpoints were then determined in both in vivo and in vitro uranyl acetate (UA) exposure cases. RESULTS: Single injection of UA (0, 0.5, 1 and 2mg/kg, i.p.) caused a significant increase in blood urea nitrogen and creatinine levels. Isolated mitochondria from the UA-treated rat kidney showed a marked elevation in oxidative stress accompanied by mitochondrial membrane potential (MMP) collapse as compared to control group. Incubation of isolated kidney mitochondria with UA (50, 100 and 200µM) manifested that UA can disrupt the electron transfer chain at complex II and III that leads to induction of reactive oxygen species (ROS) formation, lipid peroxidation, and glutathione oxidation. Disturbances in oxidative phosphorylation were also demonstrated through decreased ATP concentration and ATP/ADP ratio in UA-treated mitochondria. In addition, UA induced a significant damage in mitochondrial outer membrane. Moreover, MMP collapse, mitochondrial swelling and cytochrome c release were observed following the UA treatment in isolated mitochondria. GENERAL SIGNIFICANCE: Both our in vivo and in vitro results showed that UA-induced nephrotoxicity is linked to the impairment of electron transfer chain especially at complex II and III which leads to subsequent oxidative stress.

The effect of Achillea millefolium extract on spermatogenesis of male Wistar rats.

Achillea millefolium or yarrow, a native plant in many countries, has been recognized in historical medicine, mainly because of its astringent effects. However, some aspects of the toxicity of yarrow such as possible effects on male reproductive system are not well established. In this investigation, the effects of A. millefolium L. extract on spermatogenesis in adult male wistar rats were studied. Eighty-five male Wistar rats were divided into nine experimental groups (10 in each group except the ninth group). Extract was administered at the dose of 200, 400 and 800 mg/kg/day by intraperitoneal (IP) injection or through gavage for 22 days, on every other day. Three groups were determined as sham and control groups. Five rats from each group were killed and the rest of the rats were kept for 40 days later, but with no injection, to assess the reversibility of extract effect on spermatogenesis. The results of the study showed scattered immature cells on basal membrane in seminiferous tubules at the dose of 400 mg/kg/day IP. Moreover, a significant decrease in cell accumulation and vacuolization in seminiferous tubule was seen. At the dose of 800 mg/kg, IP, thickened seminiferous tubules on basal membrane, decrease in cell accumulation in seminiferous tubule, severe disarrangement, degenerative cells and severe decrease in sperm count were seen. At the dose of 800 mg/kg/day, orally, basal membrane was thickened and the disarrangement in cells was demonstrated. As a conclusion, our results suggest that the total extract of A. millefolium L. exhibit temporary antifertile activity in adult male animals.

Influence of Aloe Vera gel on dermal wound healing process in rat.

In this topical study the influence of Aloe Vera, on the wound healing process was investigated in 63 male rats with microscopic and cell count methods. On the day of surgery a round wound, of diameter 20 mm, was created on the back of rats necks under sterile conditions. The surgery day was determined as day zero (0). Then the rats were divided randomly into control and experimental groups 1 and 2. Animals in each group were sub-divided to three smaller groups, investigated every 4, 7, and 14 days. From day 0, wound surfaces were covered with gel once daily in experimental group 1 and twice daily, for 12 h interval, in experimental group 2. Each rat received 30 g of the gel. The wound surface and healing were assessed on days 4, 7, and 14, and then a sample from the wound was prepared and investigated microscopically. The results show that the number of neutrophil, macrophage, and fibroblast cells and the wound thickness in the control group were statistically different from the experimental groups. It was found that the wound diameter thickness in the experimental group was greatly lower due to twice administration of gel and the power of wound healing was more than other groups.