Avicenna Journal of Phytomedicine
Volume:9 Issue: 6, Oct 2019

Oxidative stress is associated with the pathogenesis of brain ischemia and other neurodegenerative disorders. Previous researches have shown the antioxidant activity of Viola odorata L. In this project, we studied neuro-protective and reactive oxygen species (ROS) scavenging activities of methanol  (MeOH) extract and other fractions isolated from V. odorata in PC12 cell line in serum/glucose deprivation  (SGD) condition.

The PC12 neuronal cells were pretreated for 6 hr with MeOH extract and fractions of V. odorata (1 to 25 μg/ml) followed by 24 hr incubation under SGD condition. Cell viability was measured by Alamar Blue® assay. The level of ROS was calculated using DCFH-DA. Also, Bax/Bcl-2 protein ratio was analyzed by western blot assay.

SGD condition significantly decreased cells viability (p<0.001). Pretreatment with EtOAc (12.5 and 25 µg/ml), BuOH (12, 25, 50 µg/ml) and CH2Cl2 (1.5 µg/ml) fractions of V. odorata reduced SGD-induced cytotoxicity. MeOH extract could not increase the viability significantly. All four semi polar fractions (EtOAc, BuOH, CH2Cl2 and MeOH) decreased SGD-induced ROS production and changed Bax/Bcl-2 ratio.

V. odorata showed promising effects against SGD condition; further mechanistic and clinical studies are warranted before application of V. odorata as a neuro-protective agent.

Pumpkin (Cucurbita pepo L.) seed oil mainly consists of saturated and unsaturated fatty acids. Previously, it was reported that oral administration of pumpkin seed oil (PSO) improved hair growth in male pattern alopecia. This study aimed to evaluate hair promoting activity of topical PSO in an animal model. Male Swiss mice (25-30 g) were used. Dorsal hair of mice (2 x 2.5 cm) was gently removed. Groups were treated as follows: (A) Intact control (did not receive testosterone) (B) Testosterone solution only (5% w/v); (C) Testosterone (5%) + PSO (5%); (D) Testosterone (5%) + PSO (10%) (E) Testosterone (5%) + minoxidil (2%). Application of drugs (100 µl) was done for six days a week, for 3 weeks. Observational and microscopic examinations were performed and results of different groups were compared. Topical application of testosterone significantly (p<0.01) prevented hair growth (compared with intact control). PSO (10%) increased hair growth score after 3 weeks and histopathological findings confirmed these results. After 3 weeks of treatment, the percentage of follicles in anagen phase was 95±4.6 and 44.4±15 for intact control and testosterone-onlytreated group, respectively. These percentages for PSO (10%) and minoxidil were 75±5.3 and 91.3±4.4, respectively and they could significantly (p<0.001) reverse the effects of testosterone. In conclusion, as topical application of PSO showed hair growth promotion, it might be regarded as a promising alternative for treatment of male pattern alopecia. Also, considering its composition, free fatty acids and minor components like phytoestrogens and vitamin E may have contributed to this effect.

Burning mouth syndrome (BMS) is a debilitating disorder with few limited treatment modalities. Because of the proven association between BMS symptoms, and depression and anxiety, treatment modalities that alleviate the two latter etiologic factors can be clinically effective. Thus, owing to the antidepressant and potential analgesic effects of crocin (as an active constituent of saffron), the present study was performed to compare the effect of crocin and citalopram (as control) on BMS symptoms and depression/anxiety in patients with BMS. The present double-blind randomized clinical trial was carried out on BMS patients. Patients were randomly divided into citalopram (n=21) and crocin (n=26) groups and treated for 11 weeks. BMS symptoms (based on Visual Analysis Scale (VAS)), as well as anxiety and depression (based on Hamilton questionnaire) were evaluated at baseline and during the treatment period.  Mann-Whitney, Chi-Square test, Independent t-test, Friedman, and Spearman correlation were employed for statistical analysis. Our findings showed a significant effect for crocin on the severity of BMS symptoms, anxiety and depression in BMS patients. Crocin can be considered for treatment of BMS subjects with concurrent anxiety and/or depression.

Chronic constipation is frequently observed in postmenopausal women. An herbal combination including clover plants, Roman anis or Anisone, green anis or fennel, green raisins, Alhagi maurorum, violets, Terminalia chebula, senna and golqand has been introduced in traditional books as an effective laxative. Thus, the present study aimed to assess the effectiveness of the combined herbal capsule on chronic constipation in postmenopausal women. This triple blinded, randomized placebo-controlled trial was conducted on 64 postmenopausal women. Individuals were randomly assigned to combined the herbal capsule and placebo groups. The herbal capsule included cloves (4 units), Anise and Anison (6 units each), violet flowers (12 units), Terminalia Chebula and fresh green raisins (25 units each), senna leaves, Alhagi maurorum, and Golqand (50 units each). Constipation questionnaire and the individual’s assessment of constipation symptoms scale were used for data collection. The mean frequency of bowel movements in the herbal capsule group was significantly higher than the placebo group (mean difference=4.2; 95% confidence interval: 0.3 to 4.5; p<0.001). Straining during defecation, stool amount, incomplete defecation, sensation of obstruction during defecation, and need for manipulation to facilitate removal of stools were significantly reduced in the herbal capsule group compared to the placebo group (p<0.001). The amount and consistency of defecation was also improved in the two study groups, but a significant difference was observed between the groups (p<0.001). The mean score of constipation symptoms in the herbal capsule group was significantly reduced compared to the placebo group (-15.4; -11.5 to -19.29; p<0.001). Consumption of herbal capsules improved chronic constipation in postmenopausal women.

The present study was conducted to investigate antibacterial properties of fruit and flower of Peganum harmala. Column chromatography, followed by preparative thin layer chromatography (TLC) was used for final purification. The structure of pure alkaloids was determined using spectroscopic methods (1H-NMR, 13C-NMR, UV and MS). Smoke and extract of total alkaloids were investigated for antimicrobial activity against five different microorganisms (standards and hospital isolates). The antibacterial activity was evaluated using disc diffusion assay and minimum inhibitory concentration (MIC) was determined by serial dilution methods. Chemical investigation of the chloroform extract of ripe fruit and flower of P. harmala led to identification of three alkaloids in ripe fruit and two alkaloids in the flower and leaves of this plant. Alkaloids identified in ripe fruit were harmine, peganine (vasicine) and harmaline. Two alkaloids, harmine and peganine, were detected in the flower of P. harmala, while harmaline was only found in the ripe fruit. The total alkaloids of flower were compared with total alkaloids of ripe fruit by TLC method.  Fruits and flowers had 3.12 and 3.27% alkaloid contents, respectively. Our results showed that the alkaloids and smoke were specifically more effective on Candida albicans and Gram- positive bacteria (Micrococcus luteus and Staphylococcus aureus), while Gram- negative bacteria, especially Pseudomonas aeruginosa,were less sensitive.

Thymol, a natural aromatic monoterpene phenol derived from thymus, possesses anti-inflammatory benefits. Here, we evaluated the potential of thymol therapy in improving an animal model of ulcerative colitis. Luminal instillation of acetic acid was used to induce colitis in male Wistar rats. Treatment groups daily received prednisolone (2 mg/kg, orally) or thymol (100 mg/kg, orally) for 10 consecutive days. Then, the rats were euthanized and tissue specimens were collected for evaluation of cyclooxygenase-2 (COX-2) expression by immunohistochemistry. Furthermore, the levels of total protein, nitric oxide, myeloperoxidase, malondialdehyde, IL-1, IL-6, and TNF-α were monitored in colonic homogenates. Eventually, the relative mRNA expression of IκBα and NF-κBp65 was investigated using reverse-transcriptase PCR (RT-PCR) in colonic homogenates. Both medications could reduce the mortality rate and the clinical scores of ulcerative colitis. The COX-2 expression was significantly reduced in the colons of thymol-treated animals compared to the prednisolone group. Also, the myeloperoxidase activity, nitric oxide level and malondialdehyde intensity were decreased in the colons of thymol-treated animals to a greater extent compared to the prednisolone group. Moreover, the total protein content of guts showed significant increases in the guts of thymol-treated animals in comparison to the prednisolone group. Nonetheless, thymol significantly reduced the levels of IL-6, and IL-1 compared to prednisolone. Both medications caused a significant decrease in the mRNA level of NF-κBp65, though the mRNA level of IκBα did not show significant changes between the groups. Thymol may be a promising agent to ameliorate ulcerative colitis.

In this study, the anti-fertility effect of Corchorus olitorius leaf extract (COLE) was evaluated in adult male rats. Forty rats assigned to 4 groups of 10 rats each, were orally given COLE for 28 days. Group 1 was the control group but groups 2, 3 and 4 were administered with 250, 500 and 1000 mg/kg body weight, respectively and were considered the test groups. Blood collection from the animals was performed at the end of treatments and blood samples were used for reproductive hormone assays. Also, sperm sample quality was ascertained, and basic organs were evaluated histologically for all groups. A significant fall in relative organ weights for the testes and prostate was observed following high-dose treatment (p<0.05). Sperm sample pH, and individual sperm motility, viability, progressiveness and concentration decreased, while total abnormalities increased following high-dose (1000 mg/kg) treatment (p<0.05). Serum concentration of FSH significantly increased at 500 and 1000 mg/kg dose levels while LH and testosterone concentrations were significantly higher than control at all dose levels (p<0.05) except estrogen which was higher than control at 250 and 500 mg/kg but lower at 1000 mg/kg dose level. Control testes showed intact histological architecture with mature spermatid density of 300 cells per tubule and well differentiated Leydig cells, while those animals treated with 250 and 500 mg/kg of the extract, were without significant pathology but had average spermatid densities of 200 and 280 cells per tubule, respectively. Significant azoospermia and spermatid density of only 30 cells per tubule and prostatic degeneration were seen in the group treated with 1000 mg/kg body weight of the extract. Consumption of high amounts of C. olitorius may inhibit reproductive functions in males.

Paraquat is a herbicide with potent toxicity in humans and animals. This study aimed to evaluate the protective effects of Origanum vulgare (O. vulgare) leaf extract on the acute nephrotoxicity and renal oxidative stress caused by paraquat.

We randomly assigned forty male rats into five groups (G1-G5). The G1 was used as control; G2 only received paraquat (25 mg/kg body weight (bw)/day, po); and G3, G4 and G5 received 25 mg/kg b.w/day oral doses of paraquat and O. vulgare hydroethanolic leaf extract (200, 400, 800 mg/kg bw/day, po, respectively). After 2 weeks, superoxide dismutase (SOD), renal catalase (CAT), vitamin C levels, histopathological changes, and tumor necrosis factor-α (TNF-α) gene expression as well as serum levels of urea, creatinine (Cr), and protein carbonyl (PC) were determined.

In G2, oral administration of paraquat significantly increased (p<0.05) serum Cr, urea, PC, and renal TNF-α gene expression relative to those of the control group. Renal catalase, superoxide dismutase, and vitamin C levels were decreased significantly (p<0.05) in G2 as compared to G1. Administration of O. vulgare leaf extract not only increased the renal vitamin C, CAT, and SOD but also decreased the renal TNF-α gene expression, malondialdehyde (MDA), serum urea and creatinine in paraquat-induced nephrotoxicity in rats.

Our results show that O. vulgareleaf extract has protective effects against nephrotoxicity induced by paraquat in rats. It seems that the nephroprotective effects of O. vulgare extract may be related to its antioxidant and anti-inflammatory effects.

Phenolic compounds have been considered inhibitors of various cancers.

In this study, caffeic acid and gallic acid were appraised for their possible effects on apoptotic genes expression in a breast cancer cell line in vitro. We also evaluated ligand interaction and ligand binding with estrogen receptor alpha by molecular docking. To determine half maximal inhibitory concentration, MCF-7 cells were treated with different concentrations of caffeic acid and gallic acid by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide assay. Furthermore, morphological changes in cells and alterations in P53, Mcl-1 and P21 gene expression were studied by real-time RT-PCR. Also, protein network and different interactions between the desired genes were analyzed using GeneMANIA database.

Evaluation of cell survival by MTT assay revealed that the half-maximal inhibitory concentration values for caffeic acid and gallic acid against MCF-7 cells, were 159 and 18 µg/ml, respectively. These compounds were found to affect P53, Mcl-1 and P21 gene expression; this alteration in gene expression probably occurred along with the activation of intrinsic apoptotic signaling pathway.

Via apoptosis induction, caffeic acid and gallic acid have induce toxic effects and morphological changes in breast cancer cells, suggesting their possible future application as antitumor agents.

Zataria multiflora (Zm) has been proposed for memory enhancing in Persian traditional medicine; but to now, no study has been carried out in this field yet. The aim of this research was to study the plant effect on spatial memory in scopolamine-induced amnesia and investigate in vivo anticholinesterase effect of Zm. Aerial parts of the plant were extracted with methanol and standardized on the basis of rutin content. Male rats received three doses of Zm extract (100, 200 and 400 mg/kg, intraperitoneal (ip) for 7 days) and 30 min after the latest dose, scopolamine (1 mg/kg) was administered to animals. Learning capacity and spatial memory were studied using morris water maze (MWM) and passive avoidance test (PAT) methods. Anticholinesterase activity was studied using Ellman’s method. Physostigmine (0.3 mg/kg) and piracetam (200 mg/kg) were used as positive controls. All doses of Zm extract significantly decreased the distance and time spent to find the platform in MWM and increased the time latency in PAT test. In both MWM and PAT tests, the highest effect of Zm was observed at 200 mg/kg which was in accordance with AChE inhibitory effect of the plant. Our findings indicate that Zm has anti-amnesic effect and might improve memory deficit through anticholinesterase activity.

In the present study, we aimed to examine the effect of troxerutin treatment on levels of brain-derived neurotrophic factor (BDNF), and tumor necrosis factor-alpha (TNF-α) and interleukin-6 (IL-6), as pro-inflammatory cytokines, in the blood and hippocampus of high-fat diet (HFD) fed dams and their male offspring.

Forty virgin female Wistar rats, 3 weeks old, were divided into two groups (n=20/per group) and fed control diet (CD), or HFD for 8 weeks. After mating, pregnant animals were assigned to four subgroups including: control (CD), control+troxerutin (CD+T), high-fat diet (HFD), and HFD+troxerutin (HFD+T) groups. HFD was continued during pregnancy and lactation. Troxerutin (150 mg/kg/day, P.O.) was administered during pregnancy in the CD+T and HFD+T groups. On postnatal day (PND) 21, male offspring were separated and fed a normal diet until PND 90. Inflammatory cytokines (TNF-α, and IL-6) and BDNF levels were measured in the serum and hippocampal samples using rat-specific enzyme-linked immunosorbent assay (ELISA) kits.

Our findings showed a significant increase in the serum levels of TNF-α and IL-6, but a decrease in BDNF levels in the serum of HFD-fed dams in comparison with CD group, which was reversed by troxerutin. Moreover, troxerutin treatment, during pregnancy, significantly decreased TNF-α and IL-6 levels, but increased BDNF level in the serum and hippocampus of HFD+T offspring in comparison with HFD offspring.

These results showed that troxerutin could prevent the harmful effects of maternal HFD on their offspring through inhibition of pro-inflammatory cytokines and elevation of BDNF levels.