Protective effect of a standardized Allium jesdianum extract in an Alzheimer's disease induced rat model.

Alzheimer's disease (AD) is a complex disorder with multiple underlying mechanisms. Existing treatment options mostly address symptom management and are associated with numerous side effects. Therefore, exploring alternative therapeutic agents derived from medicinal plants, which contain various bioactive compounds with diverse pharmacological effects, holds promise for AD treatment. This study aims to assess the protective effects of the hydroalcoholic extract of Allium jesdianum on cognitive dysfunction, mitochondrial and cellular parameters, as well as genetic parameters in an intracerebroventricular Streptozotocin (icv-STZ) induced rat model of AD. Male Wistar rats were injected with a single dose of STZ (3 mg/kg, icv) to establish a sporadic AD model. A. jesdianum extract (100, 200, and 400 mg/kg/day) and donepezil (5 mg/kg/day) were orally administered for 14 days following model induction. Cognitive function was evaluated using the radial arm water maze test. Mitochondrial toxicity parameters in various brain regions (whole brain, frontal cortex, hippocampus, and cerebellum) were assessed. Gene expression analysis of miR-330, miR-132, Bax, and Bcl-2 in isolated rat brain neurons was performed using RT-qPCR. A. jesdianum extract significantly attenuated cognitive dysfunction and mitigated mitochondrial toxicity induced by icv-STZ administration. Following STZ injection, there was upregulation of Bax gene expression and downregulation of miR-330, miR-132, and Bcl-2 gene expression. Treatment with A. jesdianum extract resulted in the reversal of the expression of these microRNAs and genes, indicating its potential for improving AD and reducing neuronal apoptosis. This study demonstrates the neuroprotective capabilities of A. jesdianum against STZ-induced oxidative stress and cognitive impairment in rats, highlighting its therapeutic potential in the management of AD.

Selective toxicity of Cistanche tubulosa root extract on cancerous skin mitochondria isolated from animal model of melanoma.

INTRODUCTION: As a major public health issue, skin cancer is a leading reason of death and has resulted in significant financial and human losses globally. Numerous environmental and internal variables may both drive and exacerbate the pathophysiology of skin cancer. Marine herbs and animals, including marine sponges, cucumbers, and squirts, have been shown to have cytotoxic consequences on cancerous cells in prior research. PURPOSE: melanoma mitochondria obtained from the skin of melanoma animal models are studied in this research to see whether extracts from Cistanche tubulosa, a plant endemic to the northern coasts of the Persian Gulf, have a cytotoxic impact on them. MATERIAL AND METHOD: In this study, the mitochondria were isolated from melanoma cells via differential centrifugation and treated with various concentrations (1250, 2500 and 5000 µg/ml) of methanolic extract of C. tubulosa. Then MTT, ROS, MMP decline, mitochondrial swelling, cytochrome c release and flow cytometry assays were performed on them. RESULTS: The results of the MTT assay showed that the IC50 of C. tubulosa extract is 2500 µg/ml and C. tubulosa extract induced a selectively significant (P < 0.05) concentration-dependent decrease in the SDH activity in cancerous skin mitochondria. The ROS results also showed that all concentrations of C. tubulosa extracts significantly increased ROS production, MMP decline and the release of cytochrome c in cancer group mitochondria. The swelling of mitochondria isolated from the cancer group was significantly increased compared to the control group. In addition, the results of the apoptosis assay showed that the addition of root extract of C. tubulosa on melanoma cells increased apoptosis, while it had no effect on control non-tumour cells. DISCUSSION AND CONCLUSION: Based on these results, the presence of potentially bioactive compounds in C. tubulosa makes this Persian Gulf coastal herb a strong candidate for further molecular studies and clinical research in the field of melanoma cancer therapy.

Development of a Critical Appraisal Tool (AIMRDA) for the Peer-Review of Studies Assessing the Anticancer Activity of Natural Products: A Step towards Reproducibility.

The journal of APJCP (Asian Pacific Journal of Cancer Prevention) focuses to gather relevant and up-to-date novel information's related to cancer sciences. The research methodologies and approaches adopted by the researcher are prone to variation which may be desirable in the context of novel scientific findings however, the reproducibility for these studies needs to be unified and assured. The reproducibility issues are highly concerned when preclinical studies are reported in cancer, for natural products in particular. The natural products and medicinal plants are prone to a wide variation in terms of phytochemistry and phyto-pharmacology, ultimately affecting the end results for cancer studies. Hence the need for specific guidelines to adopt a best-practice in cancer research are utmost essential. The current AIMRDA guidelines aims to develop a consensus-based tool in order to enhance the quality and assure the reproducibility of studies reporting natural products in cancer prevention. A core working committee of the experts developed an initial draft for the guidelines where more focus was kept for the inclusion of specific items not covered in previous published tools. The initial draft was peer-reviewed, experts-views provided, and improved by a scientific committee comprising of field research experts, editorial experts of different journals, and academics working in different organization worldwide. The feedback from continuous online meetings, mail communications, and webinars resulted a final draft in the shape of a checklist tool, covering the best practices related to the field of natural products research in cancer prevention and treatment. It is mandatory for the authors to read and follow the AIMRDA tool, and be aware of the good-practices to be followed in cancer research prior to any submission to APJCP. Though the tool is developed based on experts in the field, it needs to be further updated and validated in practice via implementation in the field.

Selenium and L-carnitine protects from valproic acid-Induced oxidative stress and mitochondrial damages in rat cortical neurons.

Oxidative stress and mitochondrial dysfunction have been associated with valproic acid (VPA) induced neurotoxicity. Mitochondria are vulnerable to oxidative damage and are also a major source of superoxide free radicals. Therefore, the need for mitochondrial protective and antioxidant agents for reducing valporic acid toxicity in central nerve system (CNS) is essential. In the present study, we investigated the potential beneficial effects of sodium selenite (SS) and L-carnitine (LC) against valproic acid -induced oxidative stress and mitochondrial dysfunction in isolated rat cortical neurons. Valproic acid (50, 100 and 200 µM) treatment caused a significant decrease in cellular viability, which was accompanied by increases in reactive oxygen species (ROS) generation, GSSG and GSH content, lipid peroxidation and lysosomal and mitochondrial damages. Sodium selenite (1 µM) and L-carnitine (1 mM) pretreatment attenuated valproic acid-induced decrease in cell viability. In addition, sodium selenite (1 µM) and L-carnitine (1 mM) pretreatment significantly protected against valproic acid-induced raise in oxidative stress, mitochondrial and lysosomal dysfunction, lipid peroxidation levels and depletion of GSH content. Our results in the current study provided insights into the protective mechanism by L-carnitine and sodium selenite, which is liked, to neuronal ROS generation and mitochondrial and lysosomal damages.

Selective Toxicity Effect of Fatty Acids Omega-3, 6 and 9 Combination on Glioblastoma Neurons through their Mitochondria.

Glioblastoma (GBM) is one of the most common malignant tumors of the central nervous system that occurs in the brain and is a deadly disease. Despite the different approaches to the treatment of this malignancy, the discovery of new compounds with anti-cancer effects seems necessary. In this study, the selective toxicity effects of omega 3, 6 and 9 combinations on mitochondria isolated from U87MG human glioma cells and also human embryonic kidney 293 cells (HEK293) as normal control were investigated. The results indicated that the omega 3, 6 and 9 combinations significantly reduced succinate dehydrogenase (SDH) activity only in mitochondria isolated from U87MG human glioma cells. Additionally, exposure of mitochondria isolated from U87MG human glioma cells to this combination was associated with a selective increase in the level of reactive oxygen species (ROS), the collapse of the mitochondrial membrane potential (MMP), mitochondrial swelling and cytochrome c release. However, these effects were not observed in mitochondria isolated from HEK293 cells (as a normal group). According to results, it is proposed that the combination of omega 3, 6 and 9 could induce toxicity in U87MG human glioma cells through their mitochondria. This combination can be helpful as a complementary therapy in patients with GBM.

Mitochondrial protective and antioxidant agents protect toxicity induced by depleted uranium in isolated human lymphocytes.

Depleted uranium (DU) is a by-product of the enrichment procedure of natural uranium. During production and usage, uranium may be released into the environment due to failure to follow standard procedures, thus causing environmental pollution. In this study, toxicity effects of uranium (VI) and protective role of mitochondrial permeability transition pore sealing and antioxidant agents studied by isolated human lymphocytes. Human lymphocytes were exposed to different concentrations (0.1, 0.5, 1, 2 and 5 mM) of DU for 6 h and cytotoxicity was measured by trypan blue assay. The mechanistic parameters were assessed after 1, 2 and 3 h of lymphocyte treatment with 1/2 IC506h (0.3 mM), IC506h (0.8 mM) and 2 IC506h (1.6 mM) of DU. The reactive oxygen species (ROS), lysosomal membrane destabilization, mitochondrial membrane potential (MMP), lipid peroxidation, GSH and GSSG levels on human lymphocytes exposed to UA, were measured. The results indicate that toxicity of U (VI) was concentration dependent on human lymphocytes. Also, U (VI) induced ROS production, MMP reduction, lysosomal membrane destabilization and lipid peroxidation in human lymphocytes. In U (VI) treated lymphocytes, decrease in intracellular GSH and raise in extracellular GSSG levels were observed. We report that mitochondrial permeability transition (MPT) pore sealing and antioxidant agents, have the capacity significantly to prevents, mitochondrial toxicity. Thus, the inhibition of mitochondrial oxidative stress and mitochondrial dysfunction by MPT pore sealing and antioxidant agents is associated with the inhibition of DU-induced mitochondrial damages and activation of apoptosis in lymphocytes.

Chrysin as an Anti-Cancer Agent Exerts Selective Toxicity by Directly Inhibiting Mitochondrial Complex II and V in CLL B-lymphocytes.

We investigated the effect of chrysin on isolated normal and chronic lymphocytic leukemia (CLL) B-lymphocytes and their isolated mitochondria. We report that a selective and significant increase in cytotoxicity, intracellular reactive oxygen species, mitochondrial membrane potential collapse, ADP/ATP ratio, caspase 3 activation and finally apoptosis in chrysin-treated CLL B- lymphocytes. Also we determined that chrysin selectively inhibits complex II and ATPases in cancerous mitochondria. In this study we proved that the ability of chrysin to promote apoptosis in CLL B-lymphocytes performed by selectively targeting of mitochondria. Our findings may provide a potential therapeutic approach for using chrysin to target mitochondria in CLL B-lymphocytes.

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.

Myricetin Selectively Induces Apoptosis on Cancerous Hepatocytes by Directly Targeting Their Mitochondria.

Hepatocellular carcinoma (HCC) is the third most common cause of cancer-related death. In patients for whom HCC could not be detected early, current treatments show poor tolerance and low efficacy. So, alternative therapies with good efficacy are urgently needed. The aim of this research was to evaluate the selective apoptotic effects of myricetin (MYR), a flavonoid compound, on hepatocytes and mitochondria obtained from the liver of HCC rats. In this study, HCC induced by diethylnitrosamine (DEN), as an initiator, and 2-acetylaminofluorene (2-AAF), as a promoter. To confirm the HCC induction, serum levels of alpha-fetoprotein (AFP), AST, AST and ALP and histopathological changes in the liver tissue were evaluated. Rat liver hepatocytes and mitochondria for evaluation of the selective cytotoxic effects of MYR were isolated, and mitochondrial and cellular parameters related to apoptosis signalling were then determined. Our results showed that MYR was able to induce cytotoxicity only in hepatocytes from the HCC but not from the untreated control group. Besides, MYR (12.5, 25 and 50 µM) induced a considerable increase in reactive oxygen species (ROS) level, mitochondrial swelling, mitochondrial membrane permeabilization (MMP) and cytochrome c release only in cancerous but not in untreated normal hepatocyte mitochondria. MYR selectively increased caspase-3 activation and apoptotic phenotypes in HCC, but not untreated normal hepatocytes. Finally, our finding underlines MYR as a promising therapeutic candidate against HCC and recommends the compound for further studies.

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.

Dracocephalum: novel anticancer plant acting on liver cancer cell mitochondria.

Dracocephalum kotschyi Boiss. (Labiatae) is a native Iranian medicinal plant which has been used in combination with Peganum harmala L. as a remedy for many forms of human cancer especially leukemia and gastrointestinal malignancies. Hepatocellular carcinoma (HCC) is the third leading cause of cancer-related death worldwide. In this investigation HCC was induced by a single intraperitoneal injection of diethylnitrosamine (DEN) in corn oil at 200 mg/kg body weight to rats. Two weeks after DEN administration, cancer development was promoted with dietary 2-acetylaminofluorene (2-AAF) (0.02%, w/w) for 2 weeks. Serum alpha-fetoprotein (AFP) concentration, serum alanine transaminase (ALT), aspartate transaminase (AST), and alkaline phosphatase (ALP) activities were also determined for confirmation of hepatocellular carcinoma induction. Then rat hepatocytes were isolated with collagen perfusion technique and tumoral hepatocytes were sorted by flow cytometry. Finally isolated mitochondria obtained from both tumoral and nontumoral hepatocytes were used for any probable toxic effect of Dracocephalum kotschyi ethanolic extract. Our results showed that D. kotschyi extract (250 µg/mL) induced reactive oxygen species (ROS) formation, mitochondrial membrane permeabilization (MMP), and mitochondrial swelling and cytochrome c release only in tumoral but not nontumoral hepatocyte. These findings propose Dracocephalum kotschyi as a promising candidate for future anticancer research.

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.

Protective effects of fungal ß-(1→3)-D-glucan against oxidative stress cytotoxicity induced by depleted uranium in isolated rat hepatocytes.

Previous reports suggested that certain carbohydrate polymers, such as ß-(1→3)-D-glucan, may possess free radical scavenging activity. The present study examined the free radical scavenging activity of a carbohydrate polymer, ß-(1→3)-D-glucan against oxidative stress induced by depleted uranium in isolated rat hepatocytes. Addition of U (VI) (uranyl acetate) to isolated rat hepatocytes results in reactive oxygen species (ROS) formation, rapid glutathione depletion, mitochondrial membrane potential collapse and lysosomal membrane rupture before hepatocyte lysis occurred. Our results showed that quite similar to silymarin, which is a known antioxidant and radical scavenger, tiny concentration of ß-glucan (138 nM) very successfully protected the hepatocytes against cell lysis and all oxidative stress cytotoxicity endpoints caused by depleted uranium including ROS formation, glutathione depletion, decreased mitochondrial membrane potential, lysosomal membrane rupture and caspase 3 activity increase. In conclusion, our results confirmed the antioxidant and radical scavenging activity of ß-(1→3)-D-glucan and suggested this compound and silymarin as possible drug candidates for prophylaxis and treatment against depleted uranium toxic effects.

A search for hepatoprotective activity of fruit extract of Mangifera indica L. against oxidative stress cytotoxicity.

Mango (Mangifera indica L.) and their components are commonly used in folk medicine for many curative effects. The protective effects of different concentrations of aqueous extract of Mangifera indica L. fruit (Mango Extract) (20, 50 and 100 microg/ml) and also gallic acid (100 microM) as a pure compound in the extract were examined against oxidative stress toxicity induced by cumene hydroperoxide (CHP) in isolated rat hepatocytes. The extracts and gallic acid (100 microM) protected the hepatocyte against all oxidative stress markers including cell lysis, ROS generation, lipid peroxidation, glutathione depletion, mitochondrial membrane potential decrease, lysosomal membrane oxidative damage and cellular proteolysis. Mango Extracts (20, 50 and 100 microg/ml) were more effective than gallic acid (100 microM) in protecting hepatocytes against CHP induced lipid peroxidation. On the other hand gallic acid (100 microM) acted more effective than Mango Extracts (20, 50 and 100 microg/ml) at preventing lysosomal membrane damage. In addition H(2)O(2) scavenging effect of all extracts were determined in hepatocytes and compared with gallic acid (100 microM). There were no significance differences (P<0.05) between all plant extracts and gallic acid (100 microM) in H(2)O(2) scavenging activity. These results suggest a hepatoprotective role for Mango Extract against liver injury associated with oxidative stress.

A search for hepatoprotective activity of aqueous extract of Rhus coriaria L. against oxidative stress cytotoxicity.

The protective effects of different concentrations of aqueous extract of Rhus coriaria L. fruit (75 and 100 microg/ml) and also gallic acid (100 microM) as one of its main components were examined against oxidative stress toxicity induced by cumene hydroperoxide (CHP) in isolated rat hepatocytes. Both extract concentrations and gallic acid (100 microM) significantly (P<0.05) protected the hepatocyte against all oxidative stress markers including cell lysis, ROS generation, lipid peroxidation, glutathione depletion, mitochondrial membrane potential decrease, lysosomal membrane oxidative damage and cellular proteolysis. Aqueous extracts of Rhus coriaria L. (75 and 100 microg/ml) were more effective than gallic acid (100 microM) in protecting hepatocytes against CHP induced lipid peroxidation (P<0.05). On the other hand gallic acid (100 microM) acted more effective than aqueous extracts of Rhus coriaria L. (75 and 100 microg/ml) at preventing hepatocyte membrane lysis (P<0.05). In addition H(2)O(2) scavenging effect of both extract concentrations (75 and 100 microg/ml) were determined in hepatocytes and compared with gallic acid (100 microM). Gallic acid (100 microM) was more effective than aqueous extracts of Rhus coriaria L. (75 and 100 microg/ml) at H(2)O(2) scavenging activity (P<0.05).