Amelioration of amyloid beta (Aß1-40) neurotoxicity by administration of silibinin; a behavioral and biochemical assessment.

Objectives: Alzheimer's disease (AD), the most common cause of dementia, is one of the leading causes of morbidity and death in the world. Currently, treatment mostly used to slow down the disease progression. Herbal remedies are considered by many in the community as a natural and safe treatment with fewer side effects. Silibinin, the active ingredient of Silybum marionum, has anti-oxidant, neurotrophic and neuroprotective characteristics. Therefore, here, the effect of different doses of Silibinin extract on oxidative stress and expression of neurotrophic factors was investigated. Materials and Methods: Forty eight male Wistar rats were randomly divided into sham, lesion; Aß1-40 injection, lesion-treatment; Aß1-40 injection followed by different doses of silibinin (50, 100, 200 mg / kg) through gavage and lesion-vehicle group; Aß1-40 injection + vehicle of silibinin. Morris water Maze (MWM) was done 28 days after the last treatment. Hippocampal tissue was removed for biochemical analysis. Production of nitric oxide (NO) and reactive oxygen species (ROS), expression of BDNF/VEGF and cell viability were measured using Griess, fluorimetry, Western blotting and MTT techniques. Results: Different concentrations of silibinin improved behavioral performance in animals. Higher doses of Silibinin could improve memory and learning function through MWM. Also, increasing the concentration of silibinin resulted in decreased ROS and NO production in a dose-dependent manner. Conclusion: Consequently, silibinin may act as a potential candidate for alleviating symptoms of AD.

Melatonin in cryopreservation media improves transplantation efficiency of frozen-thawed spermatogonial stem cells into testes of azoospermic mice.

BACKGROUND: Cryostorage of spermatogonial stem cells (SSCs) is an appropriate procedure for long-term storage of SSCs for fertility preservation. However, it causes damage to cellular structures through overproduction of ROS and oxidative stress. In this study, we examined the protective effect of melatonin as a potent antioxidant in the basic freezing medium to establish an optimal cryopreservation method for SSCs. METHODS: SSCs were obtained from the testes of neonatal male mice aged 3-6 days. Then, 100 µM melatonin was added to the basic freezing medium containing DMSO for cryopreservation of SSCs. Viability, apoptosis-related markers (BAX and BCL2), and intracellular ROS generation level were measured in frozen-thawed SSCs before transplantation using the MTT assay, immunocytochemistry, and flow cytometry, respectively. In addition, Western blotting and immunofluorescence were used to evaluate the expression of proliferation (PLZF and GFRα1) and differentiation (Stra8 and SCP3) proteins in frozen-thawed SSCs after transplantation into recipient testes. RESULTS: The data showed that adding melatonin to the cryopreservation medium markedly increased the viability and reduced intracellular ROS generation and apoptosis (by decreasing BAX and increasing BCL2) in the frozen-thawed SSCs (p < 0.05). The expression levels of proliferation (PLZF and GFRα1) and differentiation (Stra8 and SCP3) proteins and resumption of spermatogenesis from frozen-thawed SSCs followed the same pattern after transplantation. CONCLUSIONS: The results of this study revealed that adding melatonin as an antioxidant to the cryopreservation medium containing DMSO could be a promising strategy for cryopreservation of SSCs to maintain fertility in prepubertal male children who suffer from cancer.

The Effect of Prosopis farcta and Its Bioactive Luteolin on the Hippocampus of Mice after Induced Ischemia Reperfusion.

BACKGROUND: Ischemia plays an important role in increasing damage to the nervous system. This study aimed to evaluate the effect of Prosopis farcta (PFE) and its bioactive luteolin (Lu) and forced swimming exercise on the hippocampus of mice after induced ischemia reperfusion. METHODS: The bioactive component of PFE (Lu) was identified by HPLC. Fifty-six male mice were divided into different groups. Ischemia was induced by ligation of the common carotid artery. After mice training (swimming exercise, 8 weeks) and consuming PFE and Lu, the mice's memory ability was evaluated in the shuttle box. Histological examination was performed by Nissel staining and immunohistochemistry. RESULTS: Results showed that the ischemic mice exercised and treated with PFE and Lu had higher step-through latency (STL) compared with the nonexercised mice, and this was confirmed with time spent in the dark compartment (TDC). The number of dark cells in the ischemic group exercising and receiving PFE and Lu decreased compared to that of the other groups in the hippocampus. DCX protein expression was increased in nonexercised groups compared to that of the exercised groups and those treated with PFE and Lu, while NeuN decreased. CONCLUSIONS: Forced swimming exercise following ischemia, as well as consumption of PFE and Lu, has reduced cell death and increased neurogenesis in the hippocampus and thus may help improve memory in ischemia.

In vitro differentiation of neural stem cells derived from human olfactory bulb into dopaminergic-like neurons.

This study describes a new accessible source of neuronal stem cells that can be used in Parkinson's disease cell transplant. The human olfactory bulb contains neural stem cells (NSCs) that are responsible for neurogenesis in the brain and the replacement of damaged cellular components throughout life. NSCs are capable of differentiating into neuronal and glial cells. We isolated NSCs from the olfactory bulb of brain-death donors and differentiated them into dopaminergic neurons. The olfactory bulb tissues obtained were cultured in Dulbecco's modified Eagle's medium/nutrient mixture F12, B27 supplemented with basic fibroblast growth factor, epidermal growth factor and leukemia inhibitory factor. The NSCs and proliferation markers were assessed. The multipotentiality of olfactory bulb NSCs was demonstrated by their capacity to differentiate into neurons, oligodendrocytes and astrocytes. To generate dopaminergic neurons, olfactory bulb NSCs were differentiated in neurobasal medium, supplemented with B27, and treated with sonic hedgehog, fibroblast growth factor 8 and glial cell-derived neurotrophic factor from the 7th to the 21st day, followed by detection of dopaminergic neuronal markers including tyrosine hydroxylase and aromatic l-amino acid decarboxylase. The cells were expanded, established in continuous cell lines and differentiated into the two classical neuronal phenotypes. The percentage of co-positive cells (microtubule-associated protein 2 and tyrosine hydroxylase; aromatic l-amino acid decarboxylase and tyrosine hydroxylase) in the treated cells was significantly higher than in the untreated cells. These results illustrate the existence of multipotent NSCs in the adult human olfactory bulb that are capable of differentiating toward putative dopaminergic neurons in the presence of trophic factors. Taken together, our data encourage further investigations of the possible use of olfactory bulb NSCs as a promising cell-based therapeutic strategy for Parkinson's disease.