Anticonvulsant activities of Sambucus nigra.

OBJECTIVE: Sambucus (S) spp. is reported to possess a variety of activities and has been used in traditional medicine for many years. In spite of CNS activity of this genus, nothing is known about the anticonvulsant activity of S. nigra. MATERIALS AND METHODS: Anticonvulsant activities of methanolic extracts of bark, fruit and leaf of S. nigra at doses of 250, 500 and 1000 mg kg-1 were determined by pentylenetetrazole (PTZ) induced and maximal electroshock (MES) induced convulsions in mice. RESULTS: Normal saline treated mice showed tonic hind limb extension for a duration of 6.58 ± 1.24 s in MES model. Administration of extracts significantly and dose-dependently increased the delay of the onset of seizures and decreased significantly the duration of tonic hind limb extension. Bark extract at 500 and leaf extract at 1000 mg kg-1 gave 100% protection against seizures. They inhibited induction of convulsion and gave 100% protections against mortality. PTZ (100 mg kg-1) induced tonic seizures in all of the control mice. Pretreatment with leaf extract at 500 and 1000 mg kg-1 significantly decreased duration of tonic hind limb extension (p < 0.05 and p < 0.001). CONCLUSIONS: GABA receptors were involved in epilepsy. Reduction of mortality and increase the onset of convulsion in MES model was comparable with that of diazepam. Extracts might possibly be producing an antiepileptic action by increasing the level of GABA.

Hepatoprotective effects of zataria multiflora ethanolic extract on liver toxicity induced by cyclophosphamide in mice.

Although cyclophosphamide (CP), an alkylating agent, has been extensively used in chemotherapy, it possesses a wide spectrum of adverse effects including hepatotoxicity. This study was aimed to evaluate the protective effects of Zataria multiflora against hepatic damage induced by CP in mice.Mice were orally (gavages) pretreated with the ethanolic extract aerial parts of Zataria at doses of 50, 100, 200, or 400 mg/kg for 7 consecutive days before a single intraperitoneal injection of 200 mg/kg CP. After 24 h, animals were anesthetized, blood samples and hepatic tissues were collected and used for biochemical and histological examination.Serum levels of hepatic markers were significantly increased after only CP treated animals but restored in Zataria pretreated groups. A single dose of CP administration also markedly induced abnormality in the levels of several biomarkers associated with oxidative stress in liver tissues homogenates. However, pretreatment with Zataria significantly inhibited the abnormality of antioxidant enzymes defense system in the liver tissues. In addition, histopathological studies proved that CP causes damage to the liver, and this was evidenced by the induced dilated and congested sinusoidal space, lymphocytic infiltration between hepatocytes, portal space with moderate to severe inflammation and necrotic hepatocyte with absence of nuclei. Zataria effectively protected animals against CP-induced hepatic tissue damages.Our results reveal that Zataria produces a potent hepatoprotective role and could be a potent candidate to use concomitantly as a supplement agent against hepatotoxicity of CP for the patients undergoing chemotherapy.

Protective Effects of Melatonin against Cyclophosphamide-induced Oxidative Lung Toxicity in Mice.

This study was undertaken to evaluate the protective effects of melatonin against cyclophosphamide (CP)-induced oxidative lung toxicity in mice. Mice were pre-treated with various doses of melatonin for 7 consecutive days and were then injected with CP (200 mg/kg b. w.) 1 h after last melatonin injection. After 24 h, the mice were euthanized and their lungs were immediately harvested. Several biomarkers associated with oxidative stress in lung homogenates, such as thiobarbituric acid reactive substances (TBARs) and reduced glutathione (GSH) levels and the activity of superoxide dismutase (SOD) and catalase (CAT) were measured spectrophotometrically. A single dose of CP markedly altered the levels of these oxidative stress biomarkers in lung homogenates. However, increased lipid peroxidation, measured as TBARs, was significantly inhibited in the lung tissues of the melatonin-pretreated mice compared to the CP alone-injected group. In addition, pretreatment with melatonin also significantly restored GSH levels and SOD and CAT activities. Melatonin also effectively protected animals from CP-induced histological abnormalities in lung tissue. In conclusion, the increase in oxidative stress markers and concomitant adaptations by the antioxidant defense system indicates that oxidative stress plays an important role in CP-induced damage to the lung. Moreover, melatonin is a potent natural antioxidant that helps prevent CP-induced oxidative toxicity in mouse lung tissues. Thus, because melatonin is regarded to be a safe pineal secretory product, it may be used concomitantly as a supplement to reduce lung damage in patients undergoing chemotherapy.

The potential ameliorative effects of melatonin against cyclophosphamide-induced DNA damage in murine bone marrow cells.

OBJECTIVES: The protection afforded by melatonin, a pineal secretory product, against cyclophosphamide (CP)-induced genotoxicity in murine bone marrow cells was tested using micronuclei as an index of induced chromosomal damage. MATERIALS AND METHODS: Mice were pretreated with four different doses of melatonin (2.5, 5, 10 and 20 mg/kg by weight, b.w.) via intraperitoneal injection for five consecutive days followed by injection with CP (60 mg/kg b.w.) 1 hr after the last injection of melatonin on the fifth day. After 24 hr, mice were euthanized by cervical dislocation to evaluate micronucleated polychromatic erythrocytes (MnPCEs) and the ratio of polychromatic erythrocyte/polychromatic erythrocyte+normochromatic erythrocyte [PCE/(PCE+NCE)]. Histological examination of the bone marrow was also performed. RESULTS: Treatment with melatonin significantly reduced the number of MnPCEs induced by CP at all doses (p < 0.0001). At 20 mg/kg, melatonin had a maximum chemoprotective effect and reduced the number of MnPCEs by 6.93 fold and completely normalized the PCE/ (PCE+NCE) ratio. Administration of 20 mg/kg of melatonin led to marked proliferation and hypercellularity of immature myeloid elements after mice were treated with CP, as well as mitigated bone marrow suppression induced by CP. CONCLUSIONS: Our study revealed that melatonin has a potent antigenotoxic effect against CP-induced toxicity in mice, which may be due to the scavenging of free radicals and increased antioxidant status. Because melatonin is a safe, natural compound, it could be used concomitantly as a supplement to protect people undergoing chemotherapy.

Melatonin ameliorates oxidative stress and reproductive toxicity induced by cyclophosphamide in male mice.

The present study evaluated the efficacy of melatonin in cyclophosphamide (CP)-induced testicular injury, lipid peroxidative damage, and antioxidant enzymes status of the mice testis on the basis of biochemical and histological studies. Mice were pretreated with four different doses of melatonin (2.5, 5, 10, and, 20 mg/kg by body weight (b.w.)) via intraperitoneal injection for five consecutive days followed by injection with CP (200 mg/kg b.w.) 1 h after the last injection of melatonin on the 5th day. After 24 h, mice were euthanized, testes were immediately removed, and biochemical and histological studies were conducted. Treatment with melatonin significantly mitigates lipid peroxidation, superoxide dismutase, and catalase activity and the level of reduced glutathione content abnormality induced by CP in mice testis. Histological examination clearly demonstrates that pretreatment of melatonin prevented CP-induced spermatogenesis toxicity and spermatogenic cells reduction in mice testis. The protective effect of melatonin is likely due to the antioxidative properties of the indolamine existed in the chemical structure. Because melatonin is a safe, natural compound, it could be used concomitantly as a supplement to protect people undergoing chemotherapy against reproductive toxicity.

Potential chemoprotective effects of selenium on diazinon-induced DNA damage in rat peripheral blood lymphocyte.

The purpose of this study was to investigate the protective effects of selenium (Se) against genotoxicity induced by diazinon (DZN) in rat peripheral blood lymphocytes by micronucleus (MN) test. Animals were concurrently administered intraperitoneally with DZN in proper solvent (20 mg/kg body weight (b.w.)) and Se at three different doses (0.5, 1, and 2 mg/kg b.w.) for 30 consecutive days. The positive control group received DZN at the same dose without Se. After 24 h of last injection, 0.5 ml blood of each rat was received and cultured in culture medium for 44 h. The lymphocyte cultures were mitogenically stimulated with cytochalasin B to allow the evaluation of number of MNs in cytokinesis-blocked binucleated cells. Incubation of lymphocytes with DZN induced additional genotoxicity and is shown by increase in MNs frequency in human lymphocytes. Se at low dose of 0.5 mg/kg had a maximum effect and significantly reduced the MNs frequency in cultured lymphocytes (p < 0.0001) that reduced the frequency of MN from 12.78 ± 0.24% for DZN group to 4.40 ± 0.36. The present study revealed that Se particularly at low doses has a potent antigenotoxic effect against DZN -induced toxicity in rats, which may be due to the scavenging of free radicals and increased antioxidant status.

Diospyros lotus L. fruit extract protects G6PD-deficient erythrocytes from hemolytic injury in vitro and in vivo: prevention of favism disorder.

OBJECTIVE: The aim of this study was to evaluate the protective effect of Diospyros lotus L. fruit extract against the hemolytic damage induced by Vicia faba beans extract in both G6PD enzyme-deficient human and rat erythrocyte in vitro and in vivo. MATERIALS AND METHODS: In the former model, venous blood samples were obtained from five subjects with known G6PD deficiency and erythrocyte hemolysis induced by Vicia faba L. bean extract was asessed spectrophotometrically in the presence and absence of Diospyros lotus L. fruits extract. In the in vivo model, G6PD-deficient rats (induced by intraperitoneal injection of dehydroepiandrosterone for 35 days) pre-treated with different doses of Diospyros lotus L. (500, 750, 1000, and 1500 mg/kg, p.o for 7 days) were challenged with Vicia faba beans extract and the protective effect of the fruit extract against hemolysis was evaluated as above. RESULTS AND CONCLUSIONS: The results have shown that Diospyros lotus L. fruits extract has antioxidant activity that may protect against hemolytic damage induced by Vicia faba bean extract in both G6PD-deficient human and rat erythrocytes. The study gives a scientific basis for the efficacy of the fruit extract as used in Iran. The fact that this was shown in human erythrocytes in vitro is significant and provides a rationale for further testing in vivo in G6PD-deficient human populations.

Cytotoxicity of hydro-alcoholic extracts of Cucurbitapepo and Solanum nigrum on HepG2 and CT26 cancer cell lines.

Plants are used worldwide for the treatment of diseases, and novel drugs continue to be developed through research from plants. There are more than 20,000 species of plants used in traditional medicines, and these are all potential reservoirs for new drugs. Cucurbitapepo has been used in traditional folk medicine to treat cold and alleviate ache. Previous pharmacological tests have shown that it possesses antiviral, anti-inflammatory, and analgesic effects. Also, Solanum nigrum has been used as a diuretic and an antipyretic agent and it has also been used to cure inflammation, edema, mastitis and hepatic cancer. In this investigation, cytotoxicity of specific concentrations of hydro-alcoholic extracts of C. pepo and S. nigrum was studied on normal [Chinese hamster ovarian cells (CHO) and rat fibroblast] and cancer (HepG2 and CT26) cell lines. The cytotoxic effects and IC(50) of the extracts on the selected cell lines were studied followed by colonogenic assay method. The results showed that IC(50) of S. nigrum extract was significantly lower than that of the C. pepo extract on all four cell lines (P < 0.05). On the other hand, IC(50) of S. nigrum extract was significantly higher than the extract of Taxus baccata and Cisplatin, herbal and chemical control positive anticancer compounds, respectively, on all four cell lines (P < 0.05). As a result, it is concluded that the extract of S. nigrum has almost similar cytotoxicity to the extract of T. baccata on cancer cells.

The relationship between maternal serum magnesium level and preterm birth.

The aim of this study was to evaluate the relationship between maternal serum magnesium levels and preterm birth. This Nested case-control study carried out on 20 with preterm birth and 20 women at term birth at Imam Khomeini Hospital in Sari/Iran in 2008. The women with singleton gestation and intact fetal membrane suspected to preterm labor (case group), 10 cc blood samples were drawn into syringes and sent to laboratory of the hospital immediately. Sampling for control group was same as the case group. These samples recognized as control group just as birth occurring after week 37. Finally, serum magnesium level measured. Data analyzed using chi2, t-test and OR (Odd's Ratio). There was a relationship between the number of prenatal visits (p = 0.008) and stressful events associated with preterm birth (p < 0.02). Serum magnesium level was associated with preterm birth OR = 4.75, CI 95% = (0.48-46.91), Sensitivity, specificity, positive and negative predictive value of serum magnesium for preterm birth was 95, 50, 66.5 and 83.33%, respectively. Although, there was a correlation between serum magnesium levels and preterm birth, due to methodology of the study, a cohort study with the same cut off point and supplementation of magnesium in RTC studies is recommended.

Comparison of the cytotoxic effects of Juniperus sabina and Zataria multiflora extracts with Taxus baccata extract and Cisplatin on normal and cancer cell lines.

Isolation and identification of some potent anti-tumor compounds from medicinal plants has motivated researchers to screen different parts of plant species for the determination of anti-tumor effects. In this study, cytotoxic effects and IC(50) of specific concentrations of hydro-alcoholic extracts of fruits of Juniperus sabina and leaves of Zataria multiflora were compared with hydro-alcoholic extract of bark of Taxus baccata and Cisplatin, well-known anticancer compounds, on normal (CHO and rat fibroblast) and cancer (HepG2 and SKOV3) cell lines. The hydro-alcoholic extracts of the plants were prepared by percolation. The cytotoxic effects and IC(50) of the extracts on the cell lines were studied followed by colonogenic assay after 72 h incubation. The results showed that the extract of Juniperus sabina possesses lower IC(50) in comparison with Zataria multiflora extract on all 4 normal and cancer cell lines (P<0.05); but, IC(50) of the Juniperus sabina extract was significantly higher than the Taxus baccata extract and Cisplatin on all 4 normal and cancer cell lines (P<0.05). As a result, it is concluded that the extract of J. sabina has almost similar cytotoxicity with the extract of Taxus baccata on cancer cells.

Zinc and copper plasma concentrations in rheumatoid arthritis patients from a selected population in Iran.

The importance of trace elements in chronic inflammatory diseases is related to their cofactor role in immune system functions and in different metabolic processes in articular tissues. The aim of this study was to compare serum levels of Cu, Zn and Zn/Cu ratio in Rheumatoid Arthritis (RA) patients with healthy volunteers in Sari Rheumatology clinic, 2007. Zn and Cu plasma concentrations were assayed using atomic absorption spectrophotometery in 40 selected RA patients sera based on sex and age compared with healthy volunteers. Statistical analysis was performed by SPSS 10 software using independent sample t-test. Zn plasma content in patient group was significantly lower (p = 0.02) than that in healthy group. Also, Cu plasma content showed no differences in comparison with healthy group (p = 0.15). Results showed no correlation between Cu and Zn plasma concentrations in patient group (p = 0.946). In contrast, significant positive correlation was found between Zn and Zn/Cu ratio (p = 0.000); but decreased Zn/Cu ratio was more influenced by diminished Zn concentration. The study showed that spreading of RA in Iranian Society is related to age, sex, career and nutrition of the patients. However, consumption of Zn and Cu supplements in RA patients may be suggested by future investigations.

The study of Cu and Zn serum levels in idiopathic dilated cardiomyopathy (IDCMP) patients and its comparison with healthy volunteers.

Changes in the cupper (Cu) and zinc (Zn) concentrations have been reported previously in idiopathic dilated cardiomyopathy (IDCMP). As a result of controversial results, the aim of this study was to compare the Zn and Cu concentrations and Zn/Cu ratio of IDCMP patients to healthy volunteers. In addition, the correlation of Cu and Zn levels with age has been evaluated. The study population consisted of 18 IDCMP patients and 27 healthy volunteers. IDCMP patients had normal angiography with echocardiography supporting cardiomyopathy without pericardial and valvular diseases. Exclusion criteria were renal or hepatic insufficiency, alcohol usage, and intake of supplements containing Cu or Zn within 1 week ago. Cu and Zn levels have been assayed with atomic absorption spectrophotometry. Statistical analysis was performed with SPSS 10 software with independent sample t test for comparing the level of Cu and Zn of IDCMP patients with normal subjects and Pearson correlation to determine the correlation between numeric data. P < 0.05 was considered as significant differences. There was a trend for a lower Zn level in IDCMP patients compared to healthy volunteers. (0.97 +/- 0.25 mg/l vs. 1.12 +/- 0.42 mg/l, respectively). The mean Cu levels of IDCMP and normal subjects were 1.33 +/- 0.20 mg/l and 1.31 +/- 0.23 mg/l, respectively. There was a significant difference in Zn/Cu ratio among patients based on the NYHA classification of heart failure (P = 0.003). Age was negatively correlated with Zn levels in IDCMP group (P = 0.037) and positively with Cu levels in healthy volunteers (P = 0.012). A lower Zn level in IDCMP patients compared to healthy volunteers and specially a significant difference in Zn/Cu ratio of patients based on their NYHA classification would suggest a critical role of zinc and Cu imbalance in development of IDCMP.