Reza Sedaghat

Design of a New Line in Treatment of Experimental Rheumatoid Arthritis by Artesunate

This study was aimed to evaluate the therapeutic potency of a new antimalarial drug, artesunate, in an experimental model of rheumatoid arthritis. Collagen-induced arthritis (CIA) was induced in Lewis rats.The intraperitoneally administration of artesunate (ARS) and methotrexate (MTX) were started on day 25 postimmunization and continued until final assessment on day 35. During this period, clinical examination was intermittent. The anticollagen type II antibody (CII Ab) and nitric oxide synthesis were measured. The paws and kness were then removed for histopathology and radiography assay. The biocompatibility of ARS and MTX were assessed using fibrosarcoma cell line. Our results showed that i.p. injection of artesunate to arthritic rats induced a significant reduction in paw edema. This beneficial effect was associated with a significant decrease in anti-CII antibody response compared with untreated rats. Histopathological assessment showed reduced inflammatory cells infiltrate in joints of treated rats, and tissue edema and bone erosion in the paws were markedly reduced following ARS therapy. Moreover, our radiographic results paralleled histological findings. Cytotoxicity analysis of ARS showed greater tolerability compared with MTX. Treatment with ARS significantly diminished nitric oxide formation in treated rats compared with untreated controls. Our findings revealed the therapeutic efficacy of artesunate in experimental rheumatoid arthritis compared with a choice drug (methotrexate). This result may recommend it as a second-line drug in the treatment of rheumatoid arthritis.

Preclinical assessment of β-d-mannuronic acid (M2000) as a non-steroidal anti-inflammatory drug

Abstract Context: β-d-Mannuronic acid (M2000) has shown its therapeutic effects with the greatest tolerability and efficacy in various experimental models such as experimental autoimmune encephalomyelitis (EAE), adjuvant induced arthritis (AIA), nephrotic syndrome, and acute glomerulonephritis. Despite pharmacological effects of β-D-mannuronic acid, there have been no systematic toxicological studies on its safety so far. Objective: The study was designed to determine the acute and subchronic toxicity of β-D-mannuronic acid, an anti-inflammatory agent, in healthy male NMRI mice and Wistar rats, respectively. Materials and methods: For the acute toxicity study, the animals received orally five different single doses of β-D-mannuronic acid and were kept under observation for 14 d. In the subchronic study, 24 Wistar male rats were divided into four groups and were treated orally (gavage) once daily with test substance preparation at dose levels of 0, 50, 250, and 1250 mg/kg body weight for at least 63 consecutive days (9 weeks). Mortality, clinical signs, body weight changes, hematological and biochemical parameters, gross findings, organ weights, and histopathological determinations were monitored during the study. Results: The results of acute toxicity indicated that the LD50 of β-D-mannuronic acid is 4.6 g/kg. We found no mortality and no abnormality in clinical signs, body weight, relative organ weights, or necropsy in any of the animals in the subchronic study. Additionally, the results showed no significant difference in hematological, biochemical, and histopathological parameters in rats. Conclusions: Our results suggest that β-D-mannuronic acid is relatively safe when administered orally in animals.

Soy protein and genistein improves renal antioxidant status in experimental nephrotic syndrome

BACKGROUND AND OBJECTIVES Nephrotic syndrome is a chronic disease especially common in the childhood and adolescence. Reactive oxygen species (ROS) and free radicals have significant role in the pathogenesis of nephrotic syndrome. The aim of this study was to evaluate the effect of soy protein and genistein (main isoflavone of soybean) on renal antioxidant status of nephrotic rats. METHODS This study was done for 8 weeks on 40 adult male Sprague-Dawley rats divided into four groups of 10 rats each. Study groups included: 1-Control, 2-Nephrotic syndrome, 3-Nephrotic syndrome+soy protein diet and 4-Nephrotic syndrome+soy protein diet+genistein. Urine protein and urine creatinine were measured. After homogenization of kidney, total antioxidant capacities (TAC), activities of catalase enzyme, the concentration of malondialdehydes (MDA) and carbolynated proteins were determined spectrophotometrically. Pathological examination was done on kidneys with light microscope. Cell viability was evaluated with MTT assay on WEHI-164 fibro sarcoma cell line. The MMP2 enzyme activity was evaluated in different concentrations of genistein. RESULTS Total antioxidant capacity was significantly increased in soy genistein. Catalase activity was significantly increased in soy and soy genistein groups. Protein carbonyl and MDA were significantly lower in soy and soy genistein groups. The scores of pathological examination showed significant improvement in soy and soy genistein groups. Genistein decreased the proliferation of the WEHI-164 fibrosarcoma cell line. CONCLUSION It seems that soy protein decreases kidney damages in nephrotic syndrome. Adding genistein to soy protein causes improvements in antioxidant status of kidney tissue. Genistein decreases proliferation of cell.

Antihyperglycemic and antihyperlipidemic effect of Rumex patientia seed preparation in streptozotocin-diabetic rats

BACKGROUND AND OBJECTIVE Rumex patientia (RP) could exert beneficial health effects to ameliorate metabolic diseases. The effect of subchronic feeding of RP seeds was evaluated on serum glucose and lipid profile in streptozotocin (STZ)-diabetic rats. METHODS Wistar rats were divided into control, RP-treated control, diabetic, glibenclamide-treated diabetic, and RP-treated diabetic groups. For induction of diabetes, streptozotcin was administered at a dose of 60mg/kg. Meanwhile, RP-treated groups received RP seed powder mixed with standard pelleted food at a weight ratio of 6% for 4 weeks. Serum glucose and lipid levels were determined before the study and at 2nd and 4th weeks after the study in addition to the oxidative stress markers in hepatic tissue. RESULTS Serum glucose was significantly lower in RP-treated diabetic rats at 2nd and 4th weeks as compared to untreated diabetics (p<0.05 and p<0.01, respectively). Serum total cholesterol and triglyceride levels did not show significant reductions in RP-treated diabetic rats as compared to untreated diabetics. Serum HDL-cholesterol, however, significantly increased (p<0.05) and LDL-cholesterol showed a significant reduction (p<0.05) in RP-treated diabetic rats as compared to untreated diabetics (p<0.05). RP also attenuated the increased malondialdehyde (MDA) content and reduced activity of superoxide dismutase (SOD) in hepatic tissue. CONCLUSION Subchronic treatment of diabetic rats with RP could lessen the abnormal changes in blood glucose level and to improve lipid profile regarding HDL- and LDL-cholesterol in part due to its attenuation of lipid peroxidation in hepatic tissue.

Berberine ameliorates intrahippocampal kainate-induced status epilepticus and consequent epileptogenic process in the rat: Underlying mechanisms

Status epilepticus (SE) is a life-threatening neurologic condition, instigating epileptogenesis to transform normal brain to an epileptic condition. SE is followed by spontaneous recurrent seizures (SRS) and final development of temporal lobe epilepsy (TLE) that is resistant to treatment. Neuroprotective strategies are increasingly put forward as a promising therapy to prevent and/or manage epileptic conditions. In this study, we investigated whether berberis alkaloid, i.e. berberine (BBR), could ameliorate intrahippocampal kainate-induced SE and its consequent epileptogenic process and to explore some underlying mechanisms. BBR was daily administered at doses of 25 or 50mg/kg. Results showed that BBR treatment of kainate-microinjected rats at a dose of 50mg/kg lowered the incidence of SE and SRS. It also significantly restored hippocampal level of reactive oxygen species (ROS), glutathione (GSH), nuclear factor (erythroid-derived 2)-like 2 (Nrf2), activity of catalase and caspase 3, nuclear factor-B (NF-κB), toll-like receptor 4 (TLR4), tumor necrosis factor α (TNFα), interleukin-1β (IL-1β), neural cell adhesion molecule (NCAM), glial fibrillary acidic protein (GFAP), cathepsin D, and heme oxygenase 1 (HO-1). Additionally, BBR protected against hippocampal CA3 neuronal loss and prevented development of aberrant mossy fiber sprouting (MFS) as an essential element of chronic epileptogenic circuit. These data suggest that BBR could mitigate SE and SRS in intrahippocampal kainate model of epilepsy and exert neuroprotective effect and its influence is mainly mediated via suppression of oxidative stress, neuroinflammation, and possibly apoptosis.

Therapeutic effects of dasatinib in mouse model of multiple sclerosis

Abstract Background: Experimental autoimmune encephalomyelitis (EAE) is a mouse model for multiple sclerosis (MS). EAE is mainly mediated by adaptive and innate immune responses that lead to an inflammatory demyelination and axonal damage. Dasatinib (Sprycel) is a selective protein tyrosine kinase inhibitor with immunomodulatory properties that abrogates multiple signal transduction pathways in immune cells. In the present research, our aim was to test the therapeutic efficacy of dasatinib in experimental model of MS. Methods: We performed EAE induction in female C57BL/6 mice by myelin oligodendrocyte glycoprotein35–55 (MOG35–55) in Complete Freund’s Adjuvant (CFA) emulsion, and used dasatinib for the treatment of EAE. During the course of study, clinical evaluation was assessed, and on day 21 post-immunization blood samples were taken from the heart of mice for tumor necrosis factor-alpha (TNF-α), nitric oxide (NO) and antioxidants capacity evaluation. The mice were sacrificed and brains and cerebellums of mice were removed for histological analysis. Also for in vitro analysis, we used C6 astrocytoma cell line to evaluate the inhibitory effects of dasatinib in cell proliferation and matrix metalloproteinase-2 (MMP-2) activity. Results: Our findings demonstrated that dasatinib had beneficial effects on EAE by lower incidence, attenuation in the severity and a delay in the onset of disease. The serum level of NO and TNF-α in dasatinib treated mice was significantly lower than control mice. In vitro, dasatinib inhibited cell proliferation and MMP-2 activity. Conclusion: Dasatinib with its potential therapeutic effects and immunomodulatory properties may be recommended, after additional necessary tests and trials, for the treatment of MS.

Acetyl-l-carnitine protects dopaminergic nigrostriatal pathway in 6-hydroxydopamine-induced model of Parkinson’s disease in the rat

Parkinsons disease (PD) is a movement disorder and the second most common neurodegenerative disease worldwide in which nigrostriatal dopaminergic neurons within substantia nigra pars compacta (SNC) are lost, with clinical motor and non-motor symptoms including bradykinesia, resting tremor, rigidity, stooping posture and cognitive deficits. This study was undertaken to evaluate the neuroprotective potential of acetyl-l-carnitine (ALC) against unilateral striatal 6-hydroxydopamine (6-OHDA)-induced model of PD and to explore some involved mechanisms. In this experimental study, intrastriatal 6-OHDA-lesioned rats received ALC at doses of 100 or 200mg/kg/day for 1 week. ALC (200mg/kg) lowered apomorphine-induced rotational asymmetry and reduced the latency to initiate and the total time in the narrow beam test, reduced striatal malondialdehyde (MDA), increased catalase activity and glutathione (GSH) level, prevented reduction of nigral tyrosine hydroxylase (TH)-positive neurons and striatal TH-immunoreactivity, and lowered striatal glial fibrillary acidic protein (GFAP) and its immunoreactivity as an indicator of astrogliosis, and nuclear factor NF-kappa B and Toll-like receptor 4 (TLR4) as reliable markers of neuroinflammation. Meanwhile, ALC at both doses mitigated nigral DNA fragmentation as a valuable marker of apoptosis. The results of this study clearly suggest the neuroprotective effect of ALC in 6-OHDA-induced model of PD through abrogation of neuroinflammation, apoptosis, astrogliosis, and oxidative stress and it may be put forward as an ancillary therapeutic candidate for controlling PD.

Effect of Arnebia euchroma ointment on post-laser wound healing in rats

Abstract Introduction: Arnebia euchroma ointment has been used in Iranian Traditional Medicine for burn wound healing. The aim of this study is to evaluate wound healing efficacy of A. euchroma ointment on wounds induced after fractional CO2 laser in rats. Material and methods: In this study, after anesthetizing two bilateral burn wounds were induced on dorsal skin of the rat using fractional ablative CO2 laser. After applying laser, A. euchroma ointment, petrolatum, and silver sulfadiazine cream were used topically on wounds twice daily for 10 days. Digital photographs were captured from the wound surfaces every day. At the end of the study, two blinded dermatologists observed the photograph of 3rd, 5th, 7th, and 9th days after laser injury and assessed erythema, crusting/scabbing, epithelial confluence, and general wound appearance to determine the efficacy of wound healing. These wound-healing parameters were assessed using the 5-point scales. Results: This study showed significantly less erythema and crusting (P = 0.024 and P = 0.004, respectively) on 9th day and higher epithelial confluence and general wound appearance scores on 7th (P = 0.037 and p = 0.016, respectively) and 9th days (P = 0.008 and P = 0.016, respectively) in A. euchroma ointment compared with other groups. Conclusion: This study showed A. euchroma ointment has good healing effects on post-laser wounds in rats.

Preclinical and pharmacotoxicology evaluation of α-l-guluronic acid (G2013) as a non-steroidal anti-inflammatory drug with immunomodulatory property

Abstract Context: Therapeutic effects of α-l-guluronic acid with the greatest tolerability and efficacy (G2013) have been shown in experimental model of multiple sclerosis and other in vitro and in vivo examinations regarding α-l-guluronic acid; there are no toxicological researches on its safety although the pharmacological impacts have been recorded. Objective: This study was designed to determine the acute and sub chronic toxicity of α-l-guluronic acid in healthy male and female BALB/c mice. Materials and methods: For the acute toxicity study, the animals orally received five different single doses of α-l-guluronic acid and were kept under observation for 14 d. In the sub-chronic study, 24 male and female BALB/c mice were divided into four groups and treated daily with test substance preparation at dose levels of 0, 50, 250, and 1250 mg/kg body weight for at least 90 consecutive days. The mortality, body weight changes, clinical signs, hematological and biochemical parameters, gross findings, histopathological, and organs weight determinants were monitored during this study. Results: The results of acute toxicity indicated that the LD50 of α-l-guluronic acid is 4.8 g/kg. We found no mortality or abnormality in clinical signs, body weight, relative organs weight, or necropsy in any of the animals in the subchronic study. Additionally, the results showed no significant difference in hematological, biochemical, and histopathological parameters in rats. Conclusions: Our results suggest that α-l-guluronic acid has high safety when administered orally in animals.

The immunomodulatory effects of Pleurotus florida on cell-mediated immunity and secondary lymphoid tissues in Balb/c mice.

This study aimed to evaluate the immunomodulatory effects of Pleurotus florida on cell-mediated immunity in Balb/c mice. Aqueous extract of P. florida was isolated, lyophilized, and stored in refrigerator until application. Forty female Balb/c mice were randomly divided into seven test and one control groups. Four groups received 10, 20, 50, or 100 mg/kg/day of P. florida extract intraperitoneally, whereas three others received 200, 500, or 1000 mg/kg/day by oral administration. Moreover, the control group received the same volume of normal saline intraperitoneally. The treatments were continued once a day for 2 weeks. Then, delayed type hypersensitivity (DTH), viability of splenic cells, and histology of secondary lymphoid tissues were examined. The results showed that DTH responses were not affected by various doses of the P. florida in different routes. However, a significant decrease in splenic cells viability was observed in groups treated with intraperitoneal injection of 10, 20, and 50 mg/kg of the P. florida extract. In contrast, oral administration of 1000 mg/kg of the extract caused a significant increase in viability of splenic cells. Although atrophic changes in spleen and lymph nodes were observed in some peritoneal injected groups, some doses of oral administration were lead to hyperplastic changes. The results of this study indicated that the effects of P. florida on cellular responses depends on dose and route of administration.