The dose-dependent neuroprotective effect of norepinephrine in improving memory retrieval in an experimental model of multiple sclerosis, experimental autoimmune encephalomyelitis.

Multiple sclerosis (MS) is considered an immune-mediated inflammatory disorder that causes cognitive impairments by damaging the hippocampal tissue. Conversely, norepinephrine (NEP) has anti-inflammatory and re-myelinating properties, which improve cognitive impairments. The aim of this study was to assess the neuroprotective effects of NEP on learning and memory disorders in an experimental animal model of MS. Two guide cannulas were bilaterally implanted in the rat hippocampal CA1 regions. After recovery, the animals received 3 µl of 0.01% ethidium bromide (EtB) in each of both hippocampal regions. After three days, the rats were randomly divided into 6 groups (8 rats/group), including control, sham 1, sham 2, and three groups of NEP 0.25, 0.5, and 1 mg/kg by intrahippocampal injection. Behavioral tests (e.g. shuttle box test and open-field test) were then performed. Finally, ROS, MDA, GSH, TNF-α, IL-6, and IL-1ß concentrations in the left CA1 area, as well as using western-blot analysis, p-p38, p-JNK, p-AKT, p-ERK1/2, p-NMDA, p-AMPA, p-CREB, and BDNF proteins in the right CA1 region evaluated. The EtB injection increased ROS, MDA, TNF-α, IL-6, and IL-1ß levels, as well as p-JNK and p-P38, except all other proteins, while decreasing GSH content, as well as step-through latency and locomotor activity in sham groups compared to the control group. Conversely, NEP (0.5 and 1 mg/kg, particularly at the dose of 1 mg/kg) counterbalanced all the alterations mentioned above in comparison to the sham groups. The EtB induced learning and memory impairment; however, NEP dose-dependently restored these impairments to normal levels.

Comparison of antithyroid effects and hepatic complications of methimazole with catechin and its nanoencapsulation form in adult male rats.

OBJECTIVES: Methimazole is an antithyroid drug and is used clinically in hyperthyroidism. Liver dysfunction is one of the side effects of methimazole. Catechins are natural flavonoids and have antioxidant, antithyroid, and liver protection effects. Despite the wide range of biological properties of catechins, their effective use is limited due to poor water solubility, low stability, and low bioavailability. Catechin niosomal nanoencapsulation improves the properties of catechin and increases its antioxidant activities. METHODS: Niosomal vesicles were synthesized by the Thin Film Hydration method and their physicochemical characteristics, morphology, and percentage of trapped catechin in them were determined by dynamic light scattering (DLS), transmission electron microscopy (TEM), and spectrophotometry, respectively. In this study, 32 adult male rats were divided into 4 groups: control, 50 mg/kg methimazole, 100 mg/kg catechin, and 100 mg/kg nanocapsule niosomal form of catechin. The drugs were administered orally and the duration of treatment was 8 weeks. Then, the serum concentration of thyroid hormones and thyroid stimulating hormone (TSH) by enzyme-linked immunosorbent assay (ELISA) method, and serum liver function tests were performed using an autoanalyzer. The activities of hepatic oxidative enzymes were measured spectrophotometrically. RESULTS: Our study showed that the percentage of catechin encapsulation in the niosome was calculated to be 51%. A significant difference was observed in the catechin and encapsulated catechin treatment groups compared to the methimazole group (p <0.0001). In all three treatment groups of methimazole, catechin, and niosomal nanocapsule catechin, serum levels of TT3, TT4, FT3, FT4, body weight and daily consumption of water and food were significantly reduced compared to the control group (p <0.0001). CONCLUSIONS: The antithyroid effects of catechin and its encapsulated form were comparable to methimazole. Also, the encapsulation improved the hepatoprotective effects of catechin.

Biosynthesis and characterisation of solid lipid nanoparticles and investigation of toxicity against breast cancer cell line.

Solid lipid nanoparticles (SLNs) comprise non-toxic surface-active lipidic agents combined with appropriate ratios of drugs or essential oils. The goal of this research was to investigate the effects of the SLN synthesised using essential oils of Foeniculum vulgare on the MCF-7 breast cancer cell line. SLNs were prepared by homogenisation and ultrasound techniques and characterised by dynamic light scattering (DLS), zeta potential assessment, and transmission electron microscopy (TEM). 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay (MTT assay), flow-cytometry, and Acridine-Orange assay were employed for assessing the biological activities of the SLNs. The average particle size was 55.43 nm and the net surface charge was -29.54 ± 11.67 mV. TEM showed that the mean particle size was 33.55 nm and the synthesised SLNs had a uniform round morphology. The MTT assay showed that the prepared SLNs had high toxicity against MCF-7 cells and low toxicity against normal HUVECs cells. Flow-cytometry revealed a noteworthy rise in the subG1 peak of the cell cycle in the cancer cells treated with SLNs compared to the controls, indicating apoptosis in cancer cells. The results also showed discolouration in SLNs-treated cells, which further confirmed the induction of apoptosis and the toxicity of the SLNs against MCF-7 cells.

Vitamin B12 and estradiol benzoate improve memory retrieval through activation of the hippocampal AKT, BDNF, and CREB proteins in a rat model of multiple sclerosis.

OBJECTIVES: Multiple sclerosis (MS) causes extensive damage in the hippocampus. Vitamin B12 (vit B12) and estradiol benzoate (EB) have anti-inflammatory and re-myelination properties that make them proper in improvement of cognitive impairment. This study aimed to evaluate the effects of these compounds on learning and memory disturbances. MATERIALS AND METHODS: 77 adult male rats were implanted with stainless steel guide cannula bilaterally into the hippocampal area. The animals received 3 µl intrahippocampal EtB 0.01% and were randomly divided into eleven groups (7 rats/group). The groups included control, peanut oil (sham1), distilled water (sham 2), vit B12 (0.25, 0.5, 1 mg/kg), EB (25 and 50 mg/kg), vit B12 (0.25 mg/kg) plus EB (25 mg/kg), vit B12 (0.5 mg/kg) plus EB (25 mg/kg), and vit B12 (1 mg/kg) plus EB (50 mg/kg). The control group received intrahippocampal saline (as solvent). The locomotor activity and learning and memory functions were evaluated by open-field and shuttle-box tests, respectively. AKT, CREB, and BDNF levels were analyzed by Western blotting. RESULTS: This study has found significant deficit in passive avoidance learning, locomotor activity, as well as decrease in the levels of phosphorylated AKT, BDNF, and CREB in groups that received EtB. Vit B12 (1 mg/kg), EB (50 mg/kg), and their combination markedly improved these side effects. CONCLUSION: This study demonstrated that vit B12 and estradiol benzoate, especially in combination therapy, can be helpful in treatment of memory problems and MS-induced dysfunction through activation of the hippocampal AKT, BDNF, and CREB proteins.

Age and Dose-Dependent Effects of Alpha-Lipoic Acid on Human Microtubule- Associated Protein Tau-Induced Endoplasmic Reticulum Unfolded Protein Response: Implications for Alzheimer's Disease.

BACKGROUND: In human tauopathies, pathological aggregation of misfolded/unfolded proteins, particularly microtubule-associated protein tau (MAPT, tau) is considered to be an essential mechanism that triggers the induction of endoplasmic reticulum (ER) stress. OBJECTIVE: Here, we assessed the molecular effects of natural antioxidant alpha-lipoic acid (ALA) in human tauR406W (hTau)-induced ER unfolded protein response (ERUPR) in fruit flies. METHODS: In order to reduce hTau neurotoxicity during brain development, we used a transgenic model of tauopathy where the maximum toxicity was observed in adult flies. Then, the effects of ALA (0.001, 0.005, and 0.025% w/w of diet) in htau-induced ERUPR and behavioral dysfunctions in the ages 20 and 30 days were evaluated in Drosophila melanogaster. RESULTS: Data from expression (mRNA and protein) patterns of htau, analysis of eyes external morphology as well as larvae olfactory memory were confirmed by our tauopathy model. Moreover, the expression of ERUPR-related proteins involving Activating Transcription Factor 6 (ATF6), inositol regulating enzyme 1 (IRE1), and protein kinase RNA-like ER kinase (PERK) wase upregulated and locomotor function decreased in both ages of the model flies. Remarkably, the lower dose of ALA modified ERUPR and supported the reduction of behavioral deficits in youngest adults through the enhancement of GRP87/Bip, reduction of ATF6, downregulation of PERK-ATF4 pathway, and activation of the IRE1-XBP1 pathway. On the other hand, only a higher dose of ALA affected the ERUPR via moderation of PERK-ATF4 signaling in the oldest adults. As ALA also exerts higher protective effects on the locomotor function of younger adults when htauR406Wis expressed in all neurons (htau-elav) and mushroom body neurons (htau-ok), we proposed that ALA has age-dependent effects in this model. CONCLUSION: Taken together, based on our results, we conclude that aging potentially influences the ALA effective dose and mechanism of action on tau-induced ERUPR. Further molecular studies will warrant possible therapeutic applications of ALA in age-related tauopathies.

Alpha-lipoic acid ameliorates tauopathy-induced oxidative stress, apoptosis, and behavioral deficits through the balance of DIAP1/DrICE ratio and redox homeostasis: Age is a determinant factor.

Tauopathies belong to a heterogeneous class of neuronal diseases resulting in the metabolic disturbance. A disulfide natural compound of Alpha-Lipoic acid (ALA) has shown numerous pharmacologic, antioxidant, and neuroprotective activities under neuropathological conditions. The aim of this study was to investigate the neuroprotective effects of ALA on the tauopathy-induced oxidative disturbance and behavioral deficits. The transgenic Drosophila model of tauopathy induced by human tauR406W using GAL4/UAS system and effects of ALA (0.001, 0.005, and 0.025 % w/w of diet) on the neuropathology of tau in younger (20 days) and older (30 days) adults were investigated via biochemical, molecular, behavioral and in-situ tissue analyses. Expression of apoptosis-related proteins involving Drosophila Cyt-c-d (trigger of intrinsic apoptosis) and DrICE (effector caspase) were upregulated in both ages (20 and 30 days) and DIAP1 (caspase inhibitor) has reduced only in older model flies compared to the controls. Remarkably, all doses of ALA increased DIAP1 and glutathione (GSH) as well as reducing Cyt-c-d and lipid peroxidation (LPO) in the younger flies compared to the model flies. Moreover, the higher doses of ALA were able to decrease thiol concentrations, to increase total antioxidant capacity, and to improve the behavioral deficits (locomotor function, olfactory memory, and ethanol sensitivity) in the younger flies. On the other hand, only a higher dose of ALA was able to decrease DrICE, Cyt-c-d, LPO, and thiol as well as increasing antioxidant capacity and decreasing ethanol sensitivity (ST50, RT50) in the older flies. TUNEL assay showed that all doses of ALA could potentially increase the DIAP1/DrICE ratio and exert anti-apoptotic effects on younger, but not on the older adults. Furthermore, data obtained from the in-situ ROS assay confirmed that only a higher dose of ALA significantly decreased the ROS level at both ages. Our data showed that an effective neuroprotective dose of ALA and its mechanism of action on this model of tauopathy could potentially be influenced by longevity. Moreover, it was shown that ALA prevents apoptosis and decreases the redox homeostasis, and this partially explains the mechanism by which ALA diminishes behavioral deficits.

The role of Artemisia turanica extract on renal oxidative and biochemical markers in STZ-induced diabetes in rat.

OBJECTIVE: The aim of the current study was to investigate the protective effect of Artemisia turanica (AT) against diabetes- induced renal oxidative stress in rats. MATERIALS AND METHODS: Fifty male Wistar rats were randomly divided into five groups: control, STZ-induced diabetic rats, diabetic rats+ metformin, diabetic rats + AT extract, diabetic rats+ metformin+ AT extract. In the present study, diabetes was induced by a single-dose (55 mg/kg, ip) injection of streptozotocin (STZ). Diabetic rats were daily treated with metformin (300 mg/kg), AT extract (70 mg/kg) and metformin+ AT extract for 4 consecutive weeks. Tissue activities of superoxide dismutase (SOD) and catalase and the levels of malondialdehyde (MDA) and total thiol content were measured in kidney tissue. Serum concentrations of glucose, creatinine, and urea, as well as, lipid profile were also measured. RESULTS: STZ significantly increased the levels of glucose, triglyceride, urea and MDA compared to the control group. Total thiol content, as well as, catalase and SOD activities showed significant decreases in diabetic group when compared with the control animals. Serum glucose, triglyceride, cholesterol and renal MDA showed a significant decrease and renal total thiol and the activities of antioxidant enzymes showed significant increases in AT+STZ group compared with the diabetic group. In diabetic rats received AT+ metformin, serum LDL and HDL, renal MDA and SOD and catalase activities significantly improved compared with the diabetic rats. CONCLUSION: These findings suggested that AT extract has therapeutic effects on renal oxidative damage and lipid profile in diabetes, that possibly may be due to its antioxidant and hypolipidemic effects.

A Randomized, Triple-blind Placebo-controlled Trial to Determine the Effect of Saffron on the Serum Levels of MMP-9 and TIMP-1 in Patients with Multiple Sclerosis.

Matrix metalloproteinases (MMP)-9 facilitates the migration of T-cells to central nervous system (CNS), while tissue inhibitor of metalloproteinases-1(TIMP-1) inhibits the function of MMP-9. This study aimed to determine the appropriate treatment option for multiple sclerosis (MS). Forty-three relapsing-remitting MS (RRMS) patients were randomly divided into two groups of 22 (group A, placebo) and 21 (group B, Saffron pill) individuals. Serum samples were collected from patients' blood before using the Saffron pills/placebo pills and then after 12 months. The serum level of MMP-9 and its inhibitor, as well as TIMP-1, were measured by ELISA kits. MMP-9 serum levels noticeably decreased in patients with MS following 12 months of treatment with Saffron pills (p=0.006) while the changes were not significant before and after 12 months of treatment with placebo pills. Although the levels of TIMP-1 increased significantly after one year treating with Saffron pills (p=0.0002), a considerable difference was not observed before and after taking the placebo pills. The study finding revealed that 12-months treatment with Saffron could have a significant role in reducing the serum level of MMP-9 and increasing the serum level of TIMP-1 in RRMS patients. Therefore, modulating the serum levels of MMP-9 as an important regulator of T cell trafficking to the CNS might be a promising strategy in the treatment of MS patients.

Recent Updates in the Alzheimer's Disease Etiopathology and Possible Treatment Approaches: A Narrative Review of Current Clinical Trials.

BACKGROUND: Alzheimer's disease (AD) is the most frequent subtype of incurable neurodegenerative dementias and its etiopathology is still not clearly elucidated. OBJECTIVE: Outline the ongoing clinical trials (CTs) in the field of AD, in order to find novel master regulators. METHODS: We strictly reviewed all scientific reports from Clinicaltrials.gov and PubMed databases from January 2010 to January 2019. The search terms were "Alzheimer's disease" or "dementia" and "medicine" or "drug" or "treatment" and "clinical trials" and "interventions". Manuscripts that met the objective of this study were included for further evaluations. RESULTS: Drug candidates have been categorized into two main groups including antibodies, peptides or hormones (such as Ponezumab, Interferon ß-1a, Solanezumab, Filgrastim, Levemir, Apidra, and Estrogen), and naturally-derived ingredients or small molecules (such as Paracetamol, Ginkgo, Escitalopram, Simvastatin, Cilostazo, and Ritalin-SR). The majority of natural candidates acted as anti-inflammatory or/and anti-oxidant and antibodies exert their actions via increasing amyloid-beta (Aß) clearance or decreasing Tau aggregation. Among small molecules, most of them that are present in the last phases act as specific antagonists (Suvorexant, Idalopirdine, Intepirdine, Trazodone, Carvedilol, and Risperidone) or agonists (Dextromethorphan, Resveratrol, Brexpiprazole) and frequently ameliorate cognitive dysfunctions. CONCLUSION: The presences of a small number of candidates in the last phase suggest that a large number of candidates have had an undesirable side effect or were unable to pass essential eligibility for future phases. Among successful treatment approaches, clearance of Aß, recovery of cognitive deficits, and control of acute neuroinflammation are widely chosen. It is predicted that some FDA-approved drugs, such as Paracetamol, Risperidone, Escitalopram, Simvastatin, Cilostazoand, and Ritalin-SR, could also be used in off-label ways for AD. This review improves our ability to recognize novel treatments for AD and suggests approaches for the clinical trial design for this devastating disease in the near future.

Study on the interaction of triaryl-dihydro-1,2,4-oxadiazoles with α-glucosidase.

PURPOSE: One of the therapeutic approaches in the management of Type 2 diabetes is delaying the absorption of glucose through α-glucosidase enzymes inhibition, which can reduce the incidence of postprandial hyperglycemia. The existence of chronic postprandial hyperglycemia impaired the endogenous antioxidant defense due to inducing oxidative stress induced pancreatic ß-cell destruction through uncontrolled free radicals generation such as ROS, which in turn, leads to various macrovascular and microvascular complications. This study aimed to synthesize 2-aryl-4,6-diarylpyrimidine derivatives, screen their α-glucosidase inhibitory activity, perform kinetic and molecular docking studies. METHODS: A series of 3,4,5-triphenyl-4,5-dihydro-1,2,4-oxadiazole derivatives were synthesized and their α-glucosidase inhibitory activity was screened in vitro. Compounds 6a-k were synthesized via a two-step reaction with a yield between 65 and 88%. The structural elucidation of the synthesized derivatives was performed by different spectroscopic techniques. α-Glucosidase inhibitory activity of the oxadiazole derivatives 6a-k was evaluated against Saccharomyces cerevisiae α-glucosidase. RESULTS: Most of the synthesized compounds demonstrated α-glucosidase inhibitory action. Particularly compounds 6c, 6d and 6 k were the most active compounds with IC50 values 215 ± 3, 256 ± 3, and 295 ± 4 µM respectively. A kinetic study performed for compound 6c revealed that the compound is a competitive inhibitor of Saccharomyces cerevisiae α-glucosidase with Ki of 122 µM. The docking study also revealed that the two compounds, 6c and 6 k, have important binding interactions with the enzyme active site. CONCLUSION: The overall results of our study reveal that the synthesized compounds could be a potential candidate in the search for novel α-glucosidase inhibitors to manage the postprandial hyperglycemia incidence. Graphical abstract.

The effect of hydroalcoholic extract of Ziziphora clinopodioides L. on spatial memory and neuronal density of hippocampal CA1 region in rats with sporadic Alzheimer's disease.

OBJECTIVE: Alzheimer's disease is a neurodegenerative disorder associated with gradual loss of cognitive and memory abilities. It was shown that the hippocampus is one of the first structures in the brain that is affected by the disease. Ziziphora clinopodioides (Z. clinopodioides) is a member of Lamiaceae family and contains various substances. MATERIALS AND METHODS: In this experimental study, 72 adult male Wistar rats were used for behavioral and histopathologic studies. They were divided into nine groups included: control, negative control (Alzheimer), positive control (Alzheimer's treated with rivastigmine), aCSF (artificial cerebrospinal fluid) + ziziphora extract with doses of 200,400, and 600 mg/kg, and STZ (stereptozotocine)+ziziphora extract in 200,400,600 mg/kg doses. The injury was created with bilaterally intraventricular injection. The spatial memory was studied by passive avoidance test and neuronal density was evaluated by dissector method. To examine the histopathological lesions, Congo red and toluidine blue staining were done. Data were analyzed by ANOVA Minitab software. RESULTS: The memory index (neuronal density and passive avoidance test results) showed a significant decrease in negative control group compared to control (p≤0.001). Treatment with the hydroalcoholic extract at the doses of 400 and 600 mg/kg showed a significant increase in memory index in rats with Alzheimer's disease (p≤0.001). The effect of 200 mg/kg extract was not significantly different from that of the negative control group. The results of histological analysis indicated beta-amyloid plaques formation in the control group as compared to the negative control group while treatment with the extract at the doses of 400 and 600 mg/kg, significantly reduced beta-amyloid plaques formation. CONCLUSION: These findings suggest that the extract of Z. clinopodioides can improve Alzheimer's condition and alleviate memory and histopathologic damages; also, it decreases beta-amyloid plaques and apoptosis in CA1 region of the hippocampus.

Liver Dysfunction and Oxidative Stress in Streptozotocin-Induced Diabetic Rats: Protective Role of Artemisia Turanica.

OBJECTIVES: Oxidative stress plays a central role in diabetes-induced complications. In the present study, the protevtive effect of Artemisia turanica (A. turanica) was evaluated against diabetes-induced liver oxidative stress and dysfunction. METHODS: Fifty male Wistar rats were randomly divided into five groups: control, diabetic, diabetic + metformin, diabetic + A. turanica extract, and diabetic + A. turanica extract + metformin. Experimental diabetes was induced by a single-dose (55 mg/kg, intraperitoneally (ip)) injection of streptozotocin (STZ). Metformin (300 mg/kg) and A. turanica extract (70 mg/kg) were orally administrated three days after STZ injection for four weeks. The levels of malondialdehyde (MDA), total thiol content and superoxide dismutase (SOD) and catalase activities were measured in the liver tissue. Serum glucose concentration, aspartate aminotransferase (AST) and alanine aminotransferase (ALT) activities were also determined. RESULTS: In the diabetic group, serum glucose concentration, serum AST and ALT activities and liver MDA level were significantly higher while tissue total thiol content as well as catalase and SOD activities were lower, compared to the control group. Serum glucose in diabetic rats treated with metformin + A. turanica extract showed a significant decrease compared with the diabetic group. In all the A. turanica extract and metformin treated groups, serum ALT, tissue MDA level, total thiol content and SOD activity significantly improved compared with the diabetic rats. However, treatment of the diabetic rats only with metformin could not significantly change the activities of catalase and AST compared with the diabetic group. CONCLUSION: These findings suggested that A. turanica extract had a therapeutic effect on liver dysfuncyion and oxidative stress induced by diabetes, that may be probably due to its antioxidant and antiinflammatory effects.

The Involvement of D1 and D2 Dopamine Receptors in the Restoration Effect of Left Frontal Anodal, but not Cathodal, tDCS on Streptozocin-Induced Amnesia.

BACKGROUND: Transcranial direct current stimulation (tDCS) is a non-invasive method that improves learning and memory. In this study, the effect of tDCS on streptozotocin (STZ) induced amnesia in the presence or absence of SCH23390 (D1 dopamine receptor antagonist) and sulpiride (dopamine D2 receptor antagonist) has been investigated in male Wistar rats. METHODS: Passive avoidance memory, locomotor activity and pain perception have been assessed by step-through, open-field and hot-plate instruments, respectively. Anodal and cathodal tDCS were exerted on the left frontal cortex with an intensity of 0.2 milliamps for 20 minutes twice a day in 2 successive days. RESULTS: Our study showed that STZ at doses of 30 and 60 mg/ml/kg caused amnesia, while they did not alter locomotor activity and a higher dose of STZ induced analgesia 14 days after injection. SCH23390 (0.001 mg/mL/kg) and sulpiride (0.1 mg/mL/kg) did not alter memory formation by themselves and amnesia induced by STZ (30 and 60 mg/mL/kg), while SCH23390 restored the analgesia induced by STZ (60 mg/mL/kg). Moreover, left frontal anodal and cathodal tDCS restored memory impairment induced by STZ (30 and 60 mg/mL/kg). Also, SCH23390 and sulpiride could prohibit the anodic stimulating effect on memory impairment induced by a dose of 60 mg/ml/kg, but they did not hinder the effect of the cathodal stimulation on this phenomenon. CONCLUSION: The study showed that D1 and D2 dopamine receptors are involved in the restoration effect of left frontal anodal- but not cathodal-tDCS in STZ-induced amnesia.

Berberine hydrochloride attenuates voluntary methamphetamine consumption and anxiety-like behaviors via modulation of oxytocin receptors in methamphetamine addicted rats.

OBJECTIVE: Methamphetamine (METH) addiction is recognized as one of the major public health concerns, with no approved pharmacological agents for treatment. Berberine hydrochloride, an isoquinoline alkaloid in plants, induces antipsychotic and anxiolytic effects. Hence, we hypothesized that berberine may modulate the METH-induced rewarding effects. MATERIALS AND METHODS: In this study, three groups of rat including control (N = 10), METH + vehicle (N = 10), and METH + berberine (N = 10) were kept in separate cages one day before expriments. METH (20 mg/L) was dissolved in tap water inside a bottle, while there was only tap water in the control bottle. Two groups received free METH solutions for two weeks (up to 12 mg/kg). Afterwards, they were abstianced for three weeks. Only one group received 100 mg/kg/day of berberine. After three weeks, locomotor activity and anxiety (elevated plus maze test) were evaluated, then the two-bottles choice model was used for one week to evaluate drug preferences. Finally, the brain of rats was removed for evaluation of oxytocin receptor expression via immunofluorescence staining method. RESULTS: The results showed that METH preference was lower in the berberine + METH group during drug intake compared to the METH group (P < .05). During withdrawal, berberine reduced anxiety-like behaviors (P < .05) and decreased locomotor activity versus the METH group (P < .001). Also, berberine increased numbers of oxytocin receptors in comparison with the METH group (P < .01). CONCLUSION: Considering the modulation of oxytocin receptors, berberine may be considered as a potential therapeutic agent for METH addiction.

The potassium channel blocker, dalfampridine diminishes ouabain-induced arrhythmia in isolated rat atria.

The aim of the present experiment was to investigate the possible antiarrhythmic effects of dalfampridine in ouabain-induced arrhythmia in rats. Twenty-four male rats including the control and dalfampridine-incubated (100 µM to 10 mM) ouabain-stimulated (40 µM) groups were used. After induction of anesthesia, the atria were isolated and the time of onset of arrhythmia and asystole were recorded. The contractile force of atria was also measured. Dalfampridine at concentration of 1 mM significantly postponed the onset of arrhythmia and asystole compared to control group (p ≤ .05). Ouabain significantly increased the atrial beating rate in control group (p ≤ .05), while pretreatment of isolated atria with dalfampridine reversed this effect. Incubation of isolated atria with ouabain did not alter the contractile force in both control- and dalfampridine-treated groups (p > .05). It is concluded that dalfampridine might possess antiarrhythmic properties in reducing the atrial arrhythmias.

Myelin Protection by Ursolic Acid in Cuprizone-Induced Demyelination in Mice.

Neuronal survival in multiple sclerosis (MS) and other demyelinating diseases depends on the preservation of myelin and remyelination of axons. Myelin protection is the main purpose to decrease myelin damage in the central nervous system (CNS). Ursolic acid (UA) as a natural product in apple is suggested to protect neural cells. This study is the first to demonstrate an effect for UA on CNS myelin loss induced by cuprizone toxin. In the current study, we hypothesized that daily treatment with UA in drinking water (1 mg/mL) prevents myelin damage by 6 weeks administration of CPZ in mice pellet which lead to corpus callosum axonal demyelination. We assessed the myelin content and the number of myelinating cells in corpus callosum by FluoroMyelin and luxol fast blue staining as well as by immunostaining against MBP and Olig2. Our finding indicated that UA could decrease the extent of demyelination area and enhanced myelin stain intensity within CC and protected oligodendrocyte lineage cells against cuprizone toxin. We could conclude that myelinated structures could be protected by UA in corpus callosum, which provide favorable evidence for the possibility of application of UA in demyelinating diseases and traumatic injuries.

A Randomized Control Trial Study to Determine the Effect of Melatonin on Serum Levels of IL-1ß and TNF-α in Patients with Multiple Sclerosis.

Multiple sclerosis (MS) is the most common neurological disease that happens at a young age. MS is an inflammatory disease; associated with the demyelination of the central nervous system. Therefore, some inflammatory factors are effective in the mechanism and progression of the disease. Melatonin, as a multi-effect substance including anti-inflammatory effects, can reduce symptoms of MS in patients with a change in their inflammatory factors level. In this study, 50 MS patients who were referred to the MS Society of Markazi Province were randomly selected. All patients were treated with routine MS treatment (interferon) and were divided into control (25 placebo recipients) and treatment (25 recipients of 3 mg melatonin per day for 24 weeks) groups. Anthropometric data of patients including height, weight, and age were determined. Blood samples were collected after fasting in order to determine serum levels of interleukin 1 beta (IL-1ß) and tumor necrosis factor-alpha (TNF-α). Then, samples were immediately centrifuged for serum separation and sera were transferred to a freezer at -80°C and serum levels of these factors were determined; using ELISA kit. The results of this study showed that there was no significant difference between the control and treatment groups in terms of serum levels of TNF-α. However, the level of IL-1ß was significantly reduced in the treatment group compared to the control group, indicating that melatonin decreases this inflammatory substance. Our findings suggest a valuable strategy in the treatment of patients who suffer from MS.

Preventive effects of the aqueous extract of Cichorium intybus L. flower on ethylene glycol-induced renal calculi in rats.

OBJECTIVE: Urolithiasis remains a global problem. Despite the availability of numerous methods, no definite therapeutic agent has been yet introduced for the prevention or treatment of kidney stones. In this study, we evaluated the possible preventive effects of aqueous extract of Cichorium intybus L. (chicory) flowers on ethylene glycol-induced renal calculi in rats. MATERIALS AND METHODS: A total of 24 Wistar rats were randomly divided into four groups and were treated for 30 days. Group A received drinking tap water, while groups B, C, and D were administered with 1% ethylene glycol for induction of calcium oxalate stone formation. Rats in groups C and D received intraperitoneal injections of the aqueous extract of chicory flowers (50 and 200 mg/kg, respectively) since the first day of the experiment. The urine volume, urine pH, and urinary levels of oxalate, citrate, calcium, uric acid, and creatinine as well as serum levels of calcium, uric acid, and creatinine were measured. After 30 days, the rats' kidneys were removed and prepared for histological evaluation of calcium oxalate deposits. One-way analysis of variance (ANOVA), followed by Tukey's test, was performed, using SPSS version 20. RESULTS: The number of calcium oxalate crystals was significantly higher in group B (ethylene glycol-only treated animals), compared to group A (control), group C (50 mg/kg of aqueous extract), and group D (200 mg/kg of aqueous extract) (p<0.05). On day 30, the urine level of citrate, oxalate (p>0.05), and creatinine (p<0.05), as well as urine pH (p<0.05) decreased in groups C and D, compared to group B. Also, urine calcium level, urine uric acid (p>0.05), and urine volume (p<0.05) were higher in group D, compared to group B. In addition, the serum level of calcium, creatinine (p<0.05), and uric acid (p<0.001) decreased in groups C and D. CONCLUSION: The aqueous extract of chicory flower (50 mg/kg) could reduce the number of calcium oxalate deposits in the urine and reduce the level of serum parameters.

Involvement of IL-1ß and IL-6 in antiarrhythmic properties of atorvastatin in ouabain-induced arrhythmia in rats.

PURPOSE: Evidence show that statins possess wide beneficial cardioprotective and anti-inflammatory effects; therefore, in the present experiment, we investigated the antiarrhythmic properties of atorvastatin in ouabain-induced arrhythmia in isolated rat atria and the role of several inflammatory cytokines in this effect. MATERIALS AND METHODS: Male rats were pretreated with either of atorvastatin (10 mg/kg) or vehicle, orally once daily for 6 weeks. After induction of anesthesia, we isolated the atria and after incubation with ouabain, time of onset of arrhythmia and asystole as well as atrial beating rate and contractile force were recorded. We also measured the atrial levels of IL-1ß, IL-6, and TNF-α after the injection of ouabain to animals. RESULTS: Pretreatment with atorvastatin significantly delayed the onset of arrhythmia and asystole compared with vehicle-treated group (p < .01, p < .001, respectively). Incubation of ouabain boosted both atrial beating rate and contractile force in vehicle-treated group (p < .05), while these responses in atorvastatin-treated group were not significant (p > .05). Injection of ouabain elevated the atrial levels of IL-1ß, IL-6, and TNF-α, while pretreatment of animals with atorvastatin could reverse the ouabain-induced increase in atrial IL-1ß and IL-6 (p < .01 and p < .05, respectively). CONCLUSIONS: It is concluded that observed antiarrhythmic effects of atorvastatin might be attributed to modulation of some inflammatory cytokines, at least IL-1ß and IL-6.

Study of the Role of Dopamine Receptors in Streptozotocin-Induced Depressive-Like Behavior Using the Forced Swim Test Model.

BACKGROUND: Diabetes is one of the most common endocrine diseases characterized by hyperglycemia. It is caused by an absolute or relative insulin deficiency or an insulin function deficiency. It is one of the major risk factors of depression, with the rate of depression in diabetic patients amounting to as high as 30%. This study examined the role of dopamine receptors in streptozotocin (STZ)-induced depressive-like behavior using the forced swim test (FST). MATERIALS AND METHODS: This study was performed on 56 Wistar male rats. STZ at doses of 30 and 60 mg/kg body weight was administered via intraperitoneal (IP) route to induce diabetes and depression in rats. Thereafter, by using halobenzazepine (SCH23390) (D1 dopamine receptor antagonist) and sulpiride (D2 receptor dopamine receptor antagonist), the role of dopamine receptors in STZ-induced depression was studied. The one-way analysis of variance technique, Tukey's range test, and t-test were used to analyze the data. The P-value less than 0.05 was regarded as statistically significant. RESULTS: Our study showed that STZ at doses of 30 and 60 mg/kg, two weeks after injection, caused prolonged immobility in FST, indicating depressive-like behavior (P<0.05 and P<0.01, respectively). SCH23390 (0.001 mg/mL/kg) and sulpiride (0.1 mg/mL/kg) did not change the variables of depression in animals that received STZ (at doses of 30 and 60 mg/mL/kg) two weeks before (P>0.05). CONCLUSION: According to our study, STZ has a depressive-like behavior two weeks after injection, and dopamine receptors do not play a role in depression associated with STZ use.