Effects of standardized Cannabis sativa extract and ionizing radiation in melanoma cells in vitro.

CONTEXT: Melanoma causes the highest number of skin cancer-related deaths worldwide. New treatment methods are essential for the management of this life-threatening disease. AIMS: In this study, we investigated the efficacy of a standardized Cannabis sativa extract alone or in combination with single radiation dose (6 Gy) in B16F10 mouse melanoma cells in an extract dose-dependent manner. MATERIALS AND METHODS: C. sativa extract at three concentrations (25, 12.5, and 6.25 µg/mL) alone for 72 h or in combination with radiation (24 h incubation after the extract treatment + 48 h incubation after exposure to radiation) were evaluated for cell viability of melanoma cells using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Cells were also treated with 6.25 µg/mL extract alone for 72 h before analyzing C. sativa-induced cell death by flow cytometry. RESULTS: Administration of the extract alone or alongside radiation substantially inhibited melanoma cell viability and proliferation in the extract dose response-dependent manner. The inhibition of melanoma cell viability was paralleled by an increase in necrosis but not apoptosis when melanoma cells were treated with the extract alone. Radiation alone did not have any antiproliferative effects, and radiation also did not synergize antiproliferative effects of the extract when the extract and radiation were combined. CONCLUSION: Our data suggest that C. sativa extract may have significant health and physiological implications for the treatment of melanoma. The results of this study also indicate that B16F10 mouse melanoma cells are radioresistant. Taken together, these findings may lead to the identification of new therapeutic strategy for the management of melanoma.

Standardized Cannabis sativa extract attenuates tau and stathmin gene expression in the melanoma cell line.

OBJECTIVES: Metastasis is the main cause of death in patients with melanoma. Cannabis-based medicines are effective adjunctive drugs in cancer patients. Tau and Stathmin proteins are the key proteins in cancer metastasis. Here we have investigated the effect of a standardized Cannabis sativa extract on cell migration and Tau and Stathmin gene expression in the melanoma cell line. MATERIALS AND METHODS: In the treatment group, melanoma (B1617) was treated 48 hr with various concentrations of standardized C. sativa extract. Cells with no treatment were considered as the control group, then study was followed by Quantitative RT-Real Time PCR assay. Relative gene expression was calculated by the ΔΔct method. Migration assay was used to evaluate cancer metastasis. RESULTS: Tau and stathmin gene expression was significantly decreased compared to the control group. Cell migration was also significantly reduced compared to controls. CONCLUSION: C. sativa decreased tau and stathmin gene expression and cancer metastasis. The results may have some clinical relevance for the use of cannabis-based medicines in patients with metastatic melanoma.

Protective effect of Melissa officinalis extract against H2O2-induced oxidative stress in human vascular endothelial cells.

Melissa officinalis L. is a medicinal plant with a large variety of pharmacological effects and traditional applications. This study aimed to evaluate the protective and antioxidant activities of the extract of M. officinalis aerial parts on human umbilical vein endothelial cells (HUVECs) under oxidative stress induced by H2O2. Cells were incubated with H2O2 (0.5 mM, 2 h) after pretreatment with M. officinalis extract (25-500 µg/mL). Cell viability was evaluated by 3-(4, 5- Dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay. The concentration of hydroperoxides and ferric reducing antioxidant power (FRAP) were measured in intra- and extra-cellular fluids. Pretreatment of HUVECs with M. officinalis extract at the concentrations of 100-500 µg/mL improved the cell viability after exposure to H2O2 significantly. It also decreased hydroperoxides concentration and increased FRAP value in both intra- and extra-cellular fluids. The results revealed antioxidant and cytoprotective effects of M. officinalis against H2O2-induced oxidative stress in HUVECs. Due to the valuable antioxidant activity, this plant extract may have potential benefits for the prevention of cardiovascular diseases associated with oxidative stress.

Effects of vitamin D-fortified low fat yogurt on glycemic status, anthropometric indexes, inflammation, and bone turnover in diabetic postmenopausal women: A randomised controlled clinical trial.

BACKGROUND & AIMS: Low levels of serum 25-hydroxy vitamin D (25(OH)D) are common in type 2 diabetic patients and cause several complications particularly, in postmenopausal women due to their senile and physiological conditions. This study aimed to assess the effects of vitamin D-fortified low fat yogurt on glycemic status, anthropometric indexes, inflammation, and bone turnover in diabetic postmenopausal women. METHODS: In a randomized, placebo-controlled, double-blind parallel-group clinical trial, 59 postmenopausal women with type 2 diabetes received fortified yogurt (FY; 2000 IU vitamin D in 100 g/day) or plain yogurt (PY) for 12 weeks. Glycemic markers, anthropometric indexes, inflammatory, and bone turnover markers were assessed at baseline and after 12 weeks. RESULTS: After intervention, in FY group (vs PY group), were observed: significant increase in serum 25(OH)D and decrease of PTH (stable values in PY); significant improvement in serum fasting insulin, HOMA-IR, HOMA-B, QUICKI, and no changes in serum fasting glucose and HbA1c (significant worsening of all indexes in PY); significant improvement in WC, WHR, FM, and no change in weight and BMI (stable values in PY); significant increase of omentin (stable in PY) and decrease of sNTX (significant increase in PY). Final values of glycemic markers (except HbA1c), omentin, and bone turnover markers significantly improved in FY group compared to PY group. Regarding final values of serum 25(OH)D in FY group, subjects were classified in insufficient and sufficient categories. Glycemic status improved more significantly in the insufficient rather than sufficient category; whereas the other parameters had more amelioration in the sufficient category. CONCLUSIONS: Daily consumption of 2000 IU vitamin D-fortified yogurt for 12 weeks improved glycemic markers (except HbA1c), anthropometric indexes, inflammation, and bone turnover markers in postmenopausal women with type 2 diabetes. TRIAL REGISTRATION: www.irct.ir (IRCT2013110515294N1).

The Effects of Adiantum capillus-veneris on Wound Healing: An Experimental In Vitro Evaluation.

BACKGROUND: A lot of remedies, mostly plant based, were mentioned in the Persian old pharmacopoeias for promoting of burn and wound healing and tissue repairing. The efficacy of most of these old remedies is unexplored till now. Adiantum capillus-veneris from Adiantaceae family is one of them that was used to treating of some kinds of chronic wounds. METHODS: Methanol extract was fractionated to four different partitions that is, hexane, ethyl acetate, n-butanol, and aqueous. The potential of A. capillus-veneris fractions in wound healing or prevention of chronic wounds were evaluated through angiogenesis and fibroblast proliferation, in addition to in vitro tests for protection against damage to fibroblasts by oxygen free radicals. RESULTS: The aqueous part of A. capillus-veneris promoted significant angiogenesis (P < 0.05) through both capillary-like tubular formations and proliferation of endothelial cells in vitro. In addition, in the tests for protection against damage to fibroblasts by oxygen free radicals, aqueous and butanol fractions showed significant protective effects in the concentrations 50, and 500 µg/ml (P < 0.05) in comparison with a control group. In the toxicity testing, it showed weak irritation in the Hen's egg test chorioallantoic membrane (CAM) bioassay at the vascular level on the CAM of the chicken and no significant cytotoxicity in the MTT assays on normal human dermal fibroblasts. CONCLUSIONS: Angiogenic effects and protective effects against oxygen free radicals suggested aqueous partition of A. capillus-veneris local application for prevention of late-radiation-induced injuries after radiation therapy and healing of external wounds similar to bedsores and burns.

Preventive Effects of a Three-month Yoga Intervention on Endothelial Function in Patients with Migraine.

BACKGROUND: Migraine is a neurovascular disorder and any interventions improving endothelial function may contribute to its treatment and prevention of vascular complications like ischemic stroke. Yoga has been shown to have several beneficial effects on cardiovascular systems. However, no randomized controlled studies to date have investigated its effects on endothelial function of migraineurs. METHODS: A total of 42 women patients with migraine were enrolled and randomized into either a Yoga exercise group or a control group. The control group received only medication for 12 weeks and the Yoga group was placed in yoga training program in addition to the same medical treatment. Blood test was given from all patients in order to measure plasma levels intercellular adhesion molecule (ICAM) and vascular cell adhesion molecule (VCAM) after yoga training program. RESULTS: Totally 32 patients were participated in the final analyses (yoga: n = 18, control: n = 14). By analyzing data between yoga and control groups after the treatment period, there was a significant decreased in plasma level of VCAM in yoga group compare with the control group (15.29 ± 2.1 ng/ml vs. 21.70 ± 3.0 ng/ml, P < 0.05), whereas there was no significant difference in ICAM level between groups (19.1 ± 1.8 ng/ml vs. 20.97 ± 1.9 ng/ml P > 0.05). CONCLUSIONS: It seems that yoga exercises, as a complementary treatment beside pharmacological treatments, can be potentially an effective way of improving vascular functions in migraineurs.

Pistacia atlantica Resin Has a Dose-Dependent Effect on Angiogenesis and Skin Burn Wound Healing in Rat.

Objectives. The aim of the present study was to evaluate the effect of Pistacia atlantica resin extract on the rat skin burn wound healing. Methods. Thirty-two Wistar rats were divided into four groups and treated by vehicle, 5%, 10%, and 20% concentration of Pistacia atlantica resin extract for 14 days (G1, G2, G3, and G4, resp.). The efficacy of treatment was assessed based on reduction of burn wound size and histological and molecular characteristics. Results. α -Pinene (46.57%) was the main content of essential oil of resin. There were no statistically significant differences between groups according to wound size analysis. The mean histological wound healing scores were not statistically different. Capillary counts of G2 and G3 were significantly higher than those of the G1 (P = 0.042 and 0.032, resp.). NO concentration in wound fluids on the 5th day of study was not significantly different between groups (P = 0.468). But bFGF concentration in G2 and G3 and PDGF concentration in G3 were significantly higher in comparison to G1 (P = 0.043, 0.017, and 0.019, resp.). Conclusion. Our results revealed that Pistacia atlantica resin extract has a concentration-dependent effect on the healing of burn wounds after 14 days of treatment by increasing the concentration of bFGF and PDGF and also through improving the angiogenesis.

New mirsinane-type diterpenes from Euphorbia microsciadia Boiss. with inhibitory effect on VEGF-induced angiogenesis.

Euphorbia microsciadia (Euphorbiaceae) is a perennial plant growing in Iran. Two new cyclomyrsinol esters, 3-O-propionyl-5, 10, 14-O-triacetyl-8-O-(2'-methyl-butanoyl)-cyclomyrsinol (1) and 3, 5, 10, 14, 15-O-pentaacetyl-8-O-isobutanoyl-cyclomyrsinol (2) were isolated from the methanolic extract of its dried aerial parts. The structures were elucidated based on (13)C- and (1)H-NMR as well as 2D-NMR, IR and different MS spectra. Anti-angiogenic activity was also evaluated on vascular endothelium growth factor (VEGF)-induced angiogenesis in cultured human umbilical vein endothelial cells in vitro by assessing capillary-like tube network formation.

The effects of folic acid on markers of endothelial function in patients with type 1 diabetes mellitus.

BACKGROUND: Endothelial dysfunction is considered as a fundamental and also preventable factor in the progression of vascular complications among type 1 diabetic patient. It occurs before the clinical manifestation of the mentioned complications. The aim of this study was to evaluate the effects of folic acid on endothelial function by measurements of adhesion molecules and von Willebrand factor (vWF) in patients with type 1 diabetes in Isfahan, Iran. METHODS: This double blind, placebo-controlled crossover trial included type 1 diabetic patients aged 5-20 years old. Selected patients were randomized into two groups of A and B to receive folic acid 5 mg daily or placebo for 8 weeks. After a 2-week washout period, patients in the two groups were swapped to receive placebo or folic acid, respectively, for another 8 weeks. Blood and urine samples were taken to evaluate glycosylated hemoglobin (HbAlc), folic acid, vWF, intracellular adhesion molecule (ICAM), vascular cell adhesion molecule (VCAM), and microalbuminuria levels. Results of these measurements were compared in the two groups before and after folic acid and placebo administrations. RESULTS: Fifty five type 1 diabetic patients aged 12.1 +/- 3.4 years with diabetes duration of 3.9 +/- 2.1 years were enrolled. Mean of folic acid level in the two studied groups was increased significantly (10.1 +/- 3.8 vs. 21.2 +/- 1.1 in group A and 15.5 +/- 1.9 vs. 19.9 +/- 2.8 in group B, p < 0.05). Mean of VCAM and microalbuminuria was decreased significantly after folic acid administration in the two groups (p < 0.05). Mean of HbA1c, ICAM and vWF did not significantly change after folic acid administration in the two groups (p > 0.05). CONCLUSION: Folic acid administration decreased the level of endothelial dysfunction measured by adhesion molecules, especially VCAM and microalbuminuria. However, it did not significantly affect vWF. Further studies with larger sample size and long-term administration of folic acid are necessary for making precise decisions in this field. Key words: endothelial function, Diabetes Mellitus, folic acid.

Effect of L-arginine on circulating endothelial progenitor cells in hypercholesterolemic rabbits.

The cardiovascular disease risk factors result in endothelial cells apoptosis and injury. Recently, endothelial progenitor cells (EPCs) have been shown to participate in ongoing endothelial repair. Nitric Oxide (NO) is essential for mobilization and functional activity of EPCs. We hypothesized that L-arginine (NO precursor) supplementation may increase EPCs number in hypercholesterolemic rabbits. EPCs, plasma levels of nitrite, von Willebrand Factor (vWF) and Intima-Media thickness ratio were determined in rabbits feeding high-cholesterol diet or high-cholesterol diet with L-arginine(3% in drinking water). L-arginine supplementation had a significant effect on the nitrite levels, and EPC numbers, and inhibited vWF increment and atherosclerosis progression (p<0.05). Our results suggest that the beneficial effect of L-arginine in prevention of atherosclerosis may be due to the modulation of EPC levels.

Early prevention by L-Arginine attenuates coronary atherosclerosis in a model of hypercholesterolemic animals; no positive results for treatment.

BACKGROUND: Endothelial dysfunction (ED) is an independent predictor of cardiovascular events. ED is also a reversible disorder, and nitric oxide donors like L-arginine may promote this process. Despite the positive results from several studies, there are some studies that have shown that L-arginine administration did not improve endothelium-dependent dilation or the inflammatory state of patients. In this study the early and the late effects of L-arginine on coronary fatty streak formation and ED biomarkers were considered in hypercholesterolemic rabbits. METHODS: 36 white male rabbits randomly assigned in 3 groups. Rabbits were fed 1% high-cholesterol diet (LP group, n = 15), or high-cholesterol diet with oral L-arginine (3% in drinking water) (EP group, n = 15) or standard diet (control group, n = 6) for 4 weeks (phase I). Afterward, all animals were fed normal diet for 4 weeks (phase II). In the second phase, L-arginine was discontinued for EP group and was begun for LP group. The plasma levels of lipids, von Willebrand factor (vWF), and nitrite were compared before and after 4 and 8 weeks of experiment. Coronary fatty streak formation was measure after 4 and 8 weeks of experiment. RESULTS: The plasma levels of lipids were increased significantly in both groups of LP and EP after phase I. The hypercholesterolemia induced significant increased vWF release in LP group. The L-arginine supplementation led to significant plasma nitrite increment in EP group. The vWF in LP group was higher than other groups (p < 0.05). By the end of phase II, despite of start of L-arginine supplementation for LP group and L-arginine discontinuation in EP group, there were significantly more fatty streaks lesions in LP group coronary arteries than EP group. Furthermore, L-arginine supplementation did not result in significant nitrite increment in LP group. CONCLUSION: Early prevention by L-arginine may be helpful to prevent the ED, but our study did not suggest the treatment. It seems reasonable to consider ED-aside from control the cardiovascular risk factors in primary prevention of atherosclerosis and its clinical outcomes before development of irreversible vascular damage.

l-Arginine supplementation enhances eNOS expression in experimental model of hypercholesterolemic rabbits aorta.

INTRODUCTION: Diminished bioavailability of nitric oxide is crucial in endothelial dysfunction and the development of atherosclerosis. Several studies have found that l-arginine as a nitric oxide (NO) donor has beneficial effect in prevention of atherosclerosis, but the mechanism is not completely known. We hypothesized that increased endothelial nitric oxide synthase (eNOS) and/or decreased inducible NOS (iNOS) expressions might be involved in the preventive effects of l-arginine in hypercholesterolemic rabbits. METHODS: Seventeen male rabbits were divided randomly in two groups. They received rabbits chow supplemented with 1% cholesterol (group 1, n=8) and the other group received also l-arginine (3% in drinking water) (group 2, n=9) for 1 month. Blood samples were obtained before and after the experiment. At the end of experiment, the aortas were harvested. The serum levels of cholesterol and low-density lipoproteins (LDL) were measured. The intima/media thickness (IMT) ratio was measured and the determination of fatty streak formation was done with the aid of light microscopy. eNOS and iNOS expression in aorta were studied with immuohistochemistery. RESULTS: The IMT ratio in first group having fatty streaks was 0.287±0.15. No fatty streak lesion was detected in l-arginine-treated group. The results also indicated that eNOS expression (intensity) in aortas was significantly higher in l-arginine-treated group (group 1: 13.62±2.7 and group 2: 21.77±2.8; p<0.05), but no significant difference was observed for iNOS expression between the groups. CONCLUSION: The expression of eNOS plays an important role in the protection of the vessel wall from atherosclerosis. l-Arginine in drinking water has a beneficial effect in the enhancement of eNOS protein expression.