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Background: The anti-inflammatory effects of Cynodon dactylon have been determined in some previous studies. Nevertheless, there is no comprehensive study about immunomodulatory effects of C. dactylon. The present study was performed to investigate the modulatory effects of C. dactylonon macrophages functions of NMRI mice.

Methods: This survey was an experimental intervention study. The  study  population  consisted  of  14  male  NMRI  mice  randomly  categorized  into  3 treatment groups and one control group (each group was contained 10 mice). Mice in treatment groups received hydroalcoholic extract of C. dactylon for 3 constitutive weeksin different doses (100,200 and 400 mg/Kg- every day, orally). Control mice received PBS at the same volume. At the end of study, macrophages isolated from peritoneal cavity of mice and the neutral red uptake, respiratory burst after challenge with tetra phorbol acetate and respiratory burst after challenge with opsonized yeast were evaluated in these population. Data were analyzed using the one-way ANOVA plus Turkey’s test. P values of less than 0.05 were considered statistically significant.

Results: The results of stimulation of macrophages with ester of phorbol showed a significant increase in respiratory burst of macrophages isolated from treatment groups compared to control mice (p<0.01). Nevertheless, when macrophages challenged with opsonized yeast, there was no significant difference between groups .Moreover, the results of neutral red uptake by macrophages didn’t show any significant difference between macrophages of control and treatment groups.

Conclusion: However, the hydroalcoholic extract of C. dactylon caused a significant decrease in the production of the potentially harmful free radical, but it could not interfere with the function of macrophages after challenge with a real infection.

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Background & aim: Lately, it has been demonstrated that the signaling by the α7 nicotinic receptors produces the anti-inflammatory condition in both macrophages and T cells. Moreover, activation of macrophages and T cells play an important role in multiple sclerosis (MS).  In the present study, the therapeutic effect of nicotine on experimental autoimmune encephalomyelitis (EAE), an animal model of MS, and its effects on T-helper cells responses was evaluated.

Methods: In the present experimental study, EAE was induced by homogenised guinea pig spinal cord and complete Freund’s adjuvant in wistar rats. Animals were allocated in two therapeutic groups (n=7 per group). Treatment with nicotine (2.5 mg/kg-daily) was started in treatment group when the treatment group developed a disability score (at day 12). At the same time, the control group received only the solvent with the same program. Signs of disease were recorded daily until the day 36 when animals were sacrificed. The Splenocytes were checked for proliferation by MTT test and cytokine production by ELISA. The level of nitric oxide in serum was checked by griess test. The data was analyzed using the Student t test and Mann-Whitney U.

Results: Nicotine administration in the treatment group significantly reduced the clinical symptoms after the onset of symptoms. Simultaneously with the decrease of the level of serum nitric oxide, nicotine significantly decreased the pro-inflammatory cytokine IL-17 and IFN-γ. The levels of anti-inflammatory IL-10 were not changed significantly. Lymphocyte proliferation was significantly decreased in treatment group compared to control group

Conclusion: The results of this study indicated that nicotine had immune modulatory effects and could be used to control MS disease.

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Background & aim: Recently, the interaction of lipopolysaccharide activated by mesenchymal stem cells and neutrophils has been proven. The aim of the present study was to evaluate the effect(s) of the Green tea extract, as a widely consumed beverage on the interaction of lipopolysaccharide activated MSCs on neutrophils.

Methods: In the present experimental study, after the isolation of mesenchymal stem cells from bone marrow of rats was conducted, these cells were stimulated with 10 ng/mL LPS for 1. Then, the supernatant was discarded and washed out to remove LPS cells.  Afterwards, the cells were incubated with different concentrations of green tea (10, 100 and 500 micrograms per ml) for 24 hours. Subsequently, the mesenchymal stem cells were co-cultured with neutrophils and incubated for other 4 h. Finally, the neutrophil function was evaluated. The data were analyzed by Kruskal wallis test. P values less than 0.05 were considered statistically significant.

Results: Pro-inflammatory mesenchymal stem cells (Challenged with LPS) caused a decrease in vitality, Neutral Red-uptake and respiratory burst of activated neutrophils. Treatment of pro-inflammatory mesenchymal stem cells with higher concentration of green tea extract potentiated the decrease rate of Neutral Red-uptake and respiratory burst of activated neutrophils. Moreover, the vitality of neutrophils increased in a dose-independent manner by green tea extract. 

Conclusion:It seemed that the treatment of mesenchymal stem cells in inflammatory condition with Green tea extract may potentiate the role of mesenchymal stem cells to increase phagocytosis and the respiratory burst of neutrophils cells.

 

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Background & aim: In traditional medicine, medicinal plants containing flavonoid compounds were used for several years in the treatment of various diseases. Quercetin is one of the flavonoid composition family that has a maximum antioxidant flavonoid content. The aim of this study was to investigate the physiological features of rats peripheral blood neutrophils treated with quercetin.

 

Methods: In the present experimental study, the rats peripheral blood neutrophils was isolated and then teated with various concentrations of quercetin (0, 5, 10 and 20 µg/ml). The metabolic activity, phagocytosis capabilities, germicidal and respiratory burst of neutrophils were measured. Kruskal-Wallis test was used to compare the analysis.

 

Results: The results demonestrated that the respiratory burst of neutrophils treated by quercitine was significantly increased at minimum concentrations of 10 micrograms per ml. Simmilary, the phagocytic ability of neutrophils was significantly increased at minimum concentrations of 10 micrograms per ml. Albith, the phagocytic ability of neutrophils was increasesd in 5 micrograms per ml, howevet this finding didn’t show any significant change.Moreover, the application of quercetin at minimum concentrations of 10 micrograms per ml lead to a significant reduction in killing and survival of neutrophils in the treatment group compared to that of the control group (P<0/05).

 

Conclusion: Quercetin could be considered as a natural immunomodulator of innate responses. This might be due to the inflammation control of neutrophils.

 

 

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Background & aim: Diabetes is a metabolic disorder characterized by hyperglycemia. Hyperglycemia through enzymatic and non-enzymatic processes causes induction of spontaneous oxidation of glucose and, by stimulating the production of active oxygen and nitrogen components, leading to oxidative stress. Thus, according to the antioxidant effects of atorvastatin and zinc oxide, the aim of this study  was to investigate the combined effect of atorvastatin and zinc oxide on oxidative stress and antioxidants in diabetic rats.

Methods: In the present experimental study, 50 female Wistar rats were selected randomly and divided into five groups of 10 (n=10) including : normal control (NC), diabetic control (DC), diabetic rats treated with, atorvastatin (20mg/kg daily-orally) (DA), zinc oxide (30mg/kg daily-rally) (DZ) and combination of each drug in half-dose (daily-orally) (DAZ), were each treated separately. Diabetic rats were induced by injection of 60 mg per kg of body weight of streptozotocin (STZ) intraperitoneally.All treatments were dissolved in distilled water for four weeks. After completion of treatment (forth week), weight and blood sugar were measured and then compared with data measured  on weight and blood sugar in the weeks before the start  and second week of the study. The lipid peroxidation level (MDA), the activity of catalase (CAT) and total antioxidant capacity (TAC) as an indicator of oxidative stress were measured in the hippocampus. Data were analyzed using ANOVA and Tukey tests.

Results: Zinc oxide and atorvastatin alone rather than decrease blood sugar, reduced the complications of diabetes, including oxidative damage and combination  of both reduced levels of diabetic complications led to the significant decrease in blood glucose levels and inhibiting the animal lose weight.

Conclusion: It seemed that the combination of atorvastatin and zinc oxide have synergistic benefits to control blood sugar levels and oxidative stress, and also resulting in control  of diabetes.

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Background and aim: Zymosan-induced peritonitis model can use to study the recruitment of monocytes and neutrophils into the peritoneal cavity and to study the effects of existing and novel anti-inflammatory drugs. This study was conducted to evaluate the effects of hydroalcoholic extract of Hypericum Perforatum on acute peritonitis induced by Zymosan in NMRI mice.

Methods: Fifty male NMRI mice were randomly allocated in 10 equal gropes and treated with 0 ,100, 200 or 400 mg/kg of hydroalcoholic extract of H. Perforatum and or 10mg/kg diclofenac 1 hours before the induction of peritonitis. To induce peritonitis, each mouse intraperitoneally received 10 µg of zymosan in 0.4 ml of saline. After 48 h, the peritoneal cavity was lavaged by 5 ml of cold PBS and the isolated cells were used  to evaluate cell differential count, nitric oxide production and severity of respiratory burst. Statistical dada tests were analyzed using the Kruskal-Wallis test, Mann-Whitney and Bonferroni adjustment.

Results: The data showed that the nitric oxide and respiratory burst which was produced from exudate cells by  peritoneal lavage in mice that received H. Perforatum at  doses of 200 and 400 mg/kg or diclofenac compared to mice received normal saline were reduced. Total cell number in peritoneal cavity significantly decreased in all treatment group. However, no significant difference was observed between treatment groups with Hypericum perforatum extract. Using diclofenac or hydroalcoholic extract of H. Perforatum caused a significant decrease in the levels of TNF-α, IL-1 and IL-6. Diclofenac caused more profound reduction in the levels of TNF-α, IL-1 compared to extract. Nevertheless, the level of IL-6 was indicated a significant decrease in mice with peritonitis received hydroalcoholic extract especially in dose 400 mg/kg compared to mice with peritonitis received diclofenac.

Conclusion: In total it seems that the hydroalcholic extract of H. Perforatum may be a suitable as natural source to control inflammation caused by acute peritonitis.

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Abstract
 
Background & aim: Easy access, rapid recovery and high potency of monocyte cell therapy have led to special attention in cell therapy research. Monocytes are considered as sticky cells to the flask. Therefore, finding the appropriate isolation method that has the least damage to the cell and its function is of particular importance. The purpose of this study was to compare the functional capabilities of monocytes after isolation with three methods: lidocaine / EDTA, trypsin and cold PBS / EDTA.
 
Methods: In this experimental study, after extraction of peripheral blood mononuclear cells from Balb / c mice, cells (107 × 1 cells / ml) were incubated in RPMI culture medium in T25 culture flask for 4 hours. After incubation time, non-adherent cells (mainly lymphocytes) were separated by two rinsing and removed from the flask. Three different methods of trypsin, lidocaine and phosphate buffer saline were used for isolation of monocyte cells. After isolating the cells with each method, the functional capabilities of the monocytes were measured and compared with each other. The collected data were analyzed using one-way ANOVA.
 
Results: The amount of extraction, survival, metabolic activity, phagocytosis percentage, respiratory explosion, nitric oxide levels, and yeast potential in cells isolated by lidocaine were significantly higher than other groups. Although the use of trypsin, although it results in the removal of more cells, is cooled to the PBS method, but these cells exhibit significant physiological impairment in comparison with the Lidocaine / EDTA or PBS / EDTA method. Neutral red uptake was detected by trypsin isolated monocyte cells in comparison to the other two methods at lower levels. In comparison between cold PBS and lidocaine, it seems that there is no significant difference between the monocytes obtained from the two methods in terms of neutralization.
 
Conclusion: Compared to trypsin and PBS / EDTA, the Lidocaine / EDTA method is an appropriate method for isolating monocytes adherent to the flask, due to the extraction of more efficient and efficient cells.
 
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Abstract

 

Background & Aim: Melatonin is produced from different sources in the body. In some of the previous studies, the effects of oncostatic melatonin have been reported. The aim of this study was to evaluate the effects of melatonin on K562 cells as a model for chronic myeloid leukemia.

 

Methods: In this experimental study, 6 × 10⁴ cells of K562 cell line were treated with melatonin for 24 hours with different concentrations of 0, 50, 75, 100, 150 and 200 μM. The morphological changes of the treated cells were evaluated by inverse optical microscopy in comparison with the control sample. The effects of inhibition of melatonin growth and fecundity were measured using MTT (dimethyl thiazole diphenyl tetrazolium bromide) and neutrally. The Wright-Gimsa staining was used to evaluate the distinction. Data were analyzed using Kruskal-Wallis statistical test.

 

Results: There was with other concentrations in this cell line. The highest inhibitory effect of cellular growth was observed in high concentrations of melatonin. Maximum cytotoxic effect of melatonin on K562 cell line was observed at 200 μm. However, at 50 μm concentration, the effect of cytotoxicity of melatonin was minimal (p <0.05).

 

Conclusion: Melatonin has the potential to inhibit proliferation of K562 cancer cells in vitro.

 

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Background & aim: The anti-inflammatory effects of cabergoline have been documented in various studies. The purpose of the present study was to evaluate the effects of cabergoline administration (as a potent D2 agonist) on clinical aspects and immune responses in rheumatoid arthritis (RA) induced in Wistar rats.

Methods: In the present experimental study, the population consisted of 40 male Wistar rats with a weight range of 160 to 180 g. Animals were randomly allocated into four groups of 10, including the control group (healthy), RA group treated with PBS (100 mg/kg orally), RA group treated with cabergoline (50 µg/kg-orally) and ultimately RA group treated with Prednisolone (10 mg/kg orally). Changes in severity of disease and changes in temperature were recorded every three days. All treatments were initiated at day 7, after induction and observation of the first symptoms of foot inflammation in all rats. Finally, the serum of rats was isolated to evaluate the levels of nitric oxide (Griess assay) and myeloperoxidase (evaluation of the ability to the reduction of hydrogen peroxide). Spleen cells were isolated in sterile conditions in order to evaluate the lymphocyte proliferation rate (MTT reduction assay), the intensity of the respiratory burst (NBT reduction assay), and the potential of neutral red phagocytosis. Data were analyzed using Kruskal-Wallis statistical tests and one-way ANOVA and Tukey tests.

Results: The cabergoline drug was similar to prednisolone, which led to a reduction in the severity of the disease and the swelling of soles of the feet (p=0.15). The serum levels of nitric oxide (p=0.0001) and myeloperoxidase (p=0.0001), the intensity of the respiratory burst of splenic phagocytic cells (p=0.002) and the proliferation rate of splenic lymphocytes (p=0.02) were significantly decreased in both treatment groups. Prednisolone indicated a more profound effect than cabergoline in reducing the lymphocyte proliferative index (p=0.001), while cabergoline effectively reduced respiratory burst activity (p=0.002). Moreover, cabergoline significantly revealed a more profound effect than prednisolone in reducing the increased levels of neutral red uptake by splenic phagocytic cells (p=0.01).

Conclusion: Considering the appropriate results in the animal model, the use of cabergoline may be considered as a useful approach to control RA.

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Abstract                

Background & aim: Glycyrrhizin is one of the most important pharmacological compounds of the Licorice plant which has anti-inflammatory properties. Ethanol is one of the most damaging substances in the brain, Increasing use of this substance has increases the amount of nerve damage. Therefore the purpose of this study was to investigate the protective effects of glycyrrhizin on ethanol-damaged B92 glial cells.

 
Methods: The present experimental study was performed in 2018. B92 cells were obtained from the Pasteur institute cell bank of Iran, (1×10⁶ cell/ml) were transferred to 96 plates and incubated with various glycyrrhizin concentration (5, 10 and 25 µM) for 24 hours. At that point, the cells were exposed to absolute ethanol for 80 minutes at concentrations of 86 mM. Finally, the viability of the cells was evaluated with neutral red (NR) uptake and MTT tests.

Results: Glycyrrhizin reduced the level of the cell membrane and mitochondrial damages induced by ethanol in a dose-dependent manner, so that 10 and 25 µM concentration of glycyrrhizin reduced the membrane damage and the mitochondrial performance damage, significantly. Nevertheless, Glycyrrhizin at concentration of 5 µM didn’t show any protective role so that at this concentration the growth and proliferation of cells reduced. 

Conclusion: The results of the present study indicated that licorice extract (glycyrrhizin) had a positive effect on survival rate of the cells and improves the vital capabilities of ethanol-damaged glial cells. Hence, glycyrrhizin dose dependently can protect the B92 cells against ethanol-induced damage.

 
 
 

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Abstract                
Background & aim: Previous studies have shown that cerium oxide nanoparticles (CeO2-NPs) have high pharmacological capacity due to their antioxidant and anti-inflammatory properties. Rheumatoid arthritis is a chronic and systemic autoimmune disease of unknown etiology. The aim of this study was to determine and effect of cerium oxide nanoparticles in rheumatoid arthritis model.
 
Methods: In the present experimental study, 40 Wistar rats weighing 90 to 110 g were collected from the laboratory animal center of the Faculty of Veterinary Medicine, Urmia University. Rats were randomly divided into four equal groups of healthy, RA affected and treated with Serium Oxide Nanoparticles(30 mg/kg oral, daily) and treated with Methotrexate(1 mg/kg oral, weekly). Rheumatoid arthritis was induced by Freund's complete adjuvant injection(0.1ml). Treatment was started when the rats exposed inflammatory symptoms in the tharsus joint (day 8) and continued until day 28 of the rats’ slaughter. Data were analyzed using one-way ANOVA and Tukey tests.
 
Results: Cerium oxide nanoparticles had a good anti-inflammatory effect to reduce the severity of foot-pad inflammation in a pattern similar to methotrexate (p=0.27). The level of neutral red uptake in the peripheral blood phagocytic cell population and the blood level of myeloperoxidase in the treated and cerium oxide groups were 0.76 0 0.08 and 10.39 ± 1.99 mmol / ml, respectively. Significantly lower levels were observed in the methotrexate group (0.98 0 0.07 and 19.2 2 2.59 mmol / ml) (p<0.05). Conversely, blood levels of nitric oxide in the methotrexate treated and recipient group (137.81±12.18 μM)) exhibited a greater decrease than that of the cerium oxide nanoparticles (165.9 13 13.29 mmol). / 0> p). There was no significant difference in severity of respiratory burst (p = 0.09) and CRP (p=0.13) in both treatment groups. Most importantly, unlike methotrexate, the intensity of lymphocyte proliferation in rats with arthritis treated with cerium oxide did not decrease significantly (p=0.13).
 
Conclusion: Due to the improved clinical and experimental appearance of affected rats, it seemed that, treatment with CeO2-NPs is a promising strategy to improve the inflammation in a rat model of RA.