Anticancer effect of buddlejasaponin IV and buddlejasaponin IVa from Clinopodium umbrosum on oral cancer cells (HN-5).

PURPOSE: Due to the existence of bioactive compounds with different biological activity in the genus Clinopodium; C. umbrosum was selected to evaluate its cell toxicity. METHODS: C. umbrosum aerial parts were solvent extracted and extracts were fractionated via various chromatographic techniques, so as to obtain two pure saponins, buddlejasaponin IVa and buddlejasaponin IV. The cytotoxicity activity of the extracts and the two pure compounds on oral cancer cells (HN-5) and human umbilical vein endothelial cells (HUVECs) were investigated by 3-(4,5-Dimethylthiazol-2-Yl)-2,5-Diphenyltetrazolium Bromide (MTT) method. To evaluate the effect on apoptosis induction, HN-5 cells were treated for 24 h and studied by FITC Annexin V/PI staining using flow cytometry. Subsequently, the apoptosis pathway was studied through real-time RT PCR. Besides, the scratch test was performed to study the cell migration. RESULTS: The cytotoxic activity of petroleum ether, chloroform and methanol extracts on HN-5 oral cancer cells after 24 h of treatment was calculated with IC50 values of >250, >167 and 239.5 µg/mL, respectively. The cytotoxic findings for buddlejasaponin IVa and buddlejasaponin IV showed that buddlejasaponin IV possessed superior cytotoxicity whilst both compounds showed their cytotoxicity through the apoptotic pathway with increasing Bax/Bcl2 ratio and the level of caspase 9. Also, HN-5 cells migration was reduced with two saponin compounds. CONCLUSION: C. umbrosum possessed significant cytotoxicity on HN-5 cells and the mechanism of cytotoxicity for its two major compounds, buddlejasaponin IVa and buddlejasaponin IV, was identified as the mitochondrial pathway of apoptosis reducing the invasive potential of HN-5 cells.

Evaluation of Phytochemical, Antioxidant and Antibacterial Activity on Astragalus Chrysostachys Boiss. Roots.

Astragalus is a well-known genus in Leguminosae family that represented more than 800 species growing in Iran. Nevertheless, there are a few reports on Astragalus plants endemic to Iran. The roots of Astragalus plants are rich in saponins, flavonoids and polysaccharides that possess various pharmacological activities. In the present study, chemical components, antioxidant and antibacterial activity of Astragalus chrysostachys Boiss. roots were evaluated. For determination of phytochemicals in Astragalus chrysostachys Boiss. roots, total hydroalcoholic extract was fractionated with ethyl acetate and n-butanol. Ethyl acetate extract as a flavonoid rich extract was analyzed using vacuum liquid chromatography and preparative TLC and consequently a major flavonoid was isolated. The structure of the obtained compound was elucidated with 1D and 2D NMR experiments. Additionally, the essential oil of the roots was analyzed by GC-MS. Antioxidant activity of all extracts was evaluated by different assays. Moreover, antibacterial activities of the extracts were also investigated against 2 Gram-positive and 2 Gram-negative bacteria using Micro-dilution Broth method. Apigenin-6, 8-di-C-glucoside was detected in ethyl acetate extract for the first time in genus Astragalus. In addition, m-tolualdehyde, acetophenone, croweacin were found to be characteristics of the volatile oil of roots. Ethyl acetate extracts revealed notable antioxidant activity in DPPH scavenging assay with IC50 value of 14.6 µg/mL. Evaluation of antibacterial activity on the tested extracts showed mild activity against Gram-positive bacteria. Since there have been no reports on Astragalus chrysostachys Boiss. to date, the present data might be promising for application of this plant derivatives in phytotherapeutic practice.

An in vitro ethnopharmacological study on Prangos ferulacea: a wound healing agent.

Introduction: Traditionally Prangos ferulacea root is being used as an effective wound healing agent especially for pus-filled wounds both in human and stocks in the western north of Iran. Regarding the subject we decided to study P. ferulacea roots essential oil (PFE) for its antimicrobial and wound healing activities. Methods: The in vitro wound healing activity of PFE was evaluated in the mouse fibroblast cell line L929 using MTT assay of cell viability and cytotoxicity indices. Scratch assay as an in vitro model of wound healing assay was also conducted in this study. Moreover, the type I collagen content was used as an indicator of progress in wound healing process using Sircol collagen assay. Besides, PFE was subjected to GC/MS to identify the chemical constituents, and antimicrobical property was also evaluated against S. aureus, S. epidermidis, E. coli, P. aeruginosa,S. paratyphi and C. albicans using agar dilution method. Results: GC/MS analysis showed that the monoterpene hydrocarbones dominated in PFE, amounting to a total percentage of 95.1% with the major constituents: ß-Phellandrene (32.1%), m-Tolualdehyde (26.2%), and δ-3-carene (25.8%). PFE inhibited the growth of S. aureus and P. aeruginusa with the MIC value of 20 µg/mL. In addition, at the second day of treatment, PFE at concentrations of 4 and 16 µg/mL significantly (P<0.001) enhanced the migration rate of L929 cells by 87.05±2.4 and 63.5±0.08 %, respectively. Moreover, the collagen production by L929 cells was increased greatly (P<0.001). Conclusion: It is proposed that the excellent antimicrobial activity along with the significant increase of migration rate and collagen production by fibroblast cells might be associated with the high content and synergistic effect of the monoterpens, corroborating the traditional usage of this plant as a wound healing agent.

Evaluating Antiproliferative and Antioxidant Activity of Marrubium crassidens.

PURPOSE: Naturally occurring substances as novel drugs in cancer therapy, at all times, represent a challenge to science since medicinal plants are proving to be brilliant sources of new chemopreventive agents. METHODS: In the present study, methanol extract from aerial parts of Marrubium crassidens was assessed for its antiproliferative activity in the breast cancer cell line MCF-7 through MTT bioassay using cell viability and cytotoxicity indices. The antioxidant property of M. crassidens extract together with its phenolic and flavonoids content were evaluated, as well. RESULTS: According to data obtained in the study, M. crassidens exhibited antiproliferative activity with a gradual rise in cytotoxicty effect setting out on 240µg/mL concentration of the extract. Moreover, the RC50 value for antioxidant activity of the extract was determined as 40µg/mL and values for the total phenolic and flavonoids were calculated as 512.64mg gallic acid equivalent and 212.73mg quercetin equivalent per 100g of dry plant material. CONCLUSION: Generally, the observed antiproliferative and antioxidant properties of M. crassidens could be certified to the high amounts of phenolic and flavonoid content detected in the extract.

Treatment of Paederus Dermatitis with Sambucus ebulus Lotion.

Paederus dermatitis is an irritant contact dermatitis due to accidental contact by a beetle belonging to the genus paederus. In this study, clinical efficacies of S. ebulus fruit extract solution in patients affected by paederus dermatitis were evaluated. A randomized double-blind, prospective, placebo-controlled clinical trial was performed in 62 patients with clinical symptoms and sings of dermatitis due to paederus beetles. The patients received either a topical solution of palemolin (a 5% S. ebulus fruit extract in ethanol 70%) or ethanol 70% topical solution thrice a day. Topical hydrocortisone ointment was prescribed for all patients. Palemolin was statistically more effective in controlling of burning, pain, inflammation, drying the wound, infections and acceleration of healing than control group (p ≤ 0.05). Specially in controlling of inflammation, palemolin had more significant efficacy (p < 0.001) than control group. About 63.6% of patients in palemolin group cured during first 24 h (versus 27.4% in control groups). The problems related to lesions in 93.9% of patients were eliminated completely during 48 hours after the beginning of the treatment by palemolin (versus 65.4% in control groups). Topical 5% solution of S. ebulus fruit extract is an effective pharmaceutical preparation in treatment of paederus dermatitis.

Chemical composition and in vitro antioxidant and antibacterial activity of Heracleum transcaucasicum and Heracleum anisactis roots essential oil.

INTRODUCTION: In vitro antioxidant and antibacterial activity and volatile compositions of two Heracleum species (Apiaceae) including Heracleum transcaucasicum and Heracleum anisactis roots Essential Oil (EO) were investigated. METHODS: The volatile compositions of EOs were analyzed by GC/Mass spectroscopy. To detect the antioxidant activity of essential oils TLC-bioautography and DPPH radical scavenging assay by spectrophotometry was performed. Additionally, the antibacterial activity of two essential oils were studied and compared against four pathogenic bacteria by agar disc diffusion method and MIC values of the EOs were determined using the broth dilution method. RESULTS: Myristicin was the dominant component in both EOs. It was identified as 96.87% and 95.15% of the essential oil composition of H. transcaucasicum and H. anisactis roots, respectively. The TLC-bioautography showed antioxidant spots in both EOs and IC50 of H. anisactis and H. transcaucasicum EO was found to be 54 µg × ml (-1) and 77 µg × ml (-1), respectively. Regarding the antimicrobial assay, H. anisactis EO exhibited weak to moderate antibacterial activity against gram-positive bacteria and also Escherichia coli, whereas the essential oil from H. transcaucasicum was inactive. CONCLUSION: Based on the results from this study, both tested EOs mainly consist of myristicin. Despite the presence of myristicin with known antibacterial property, the EO from H. transcacausicum showed no antibacterial activity. Thus it is supposed that the biological activity of plants is remarkably linked to the extracts' chemical profile and intercomponents' synergistic or antagonistic effect could play a crucial role in bioactivity of EOs and other plant extracts.

Marrubium vulgare L. methanolic extract inhibits inflammatory response and prevents cardiomyocyte fibrosis in isoproterenol-induced acute myocardial infarction in rats.

INTRODUCTION: Nowadays, finding new therapeutic compounds from natural products for treatment and prevention of a variety of diseases including cardiovascular disorders is getting a great deal of attention. This approach would result in finding new drugs which are more effective and have fewer side effects than the conventional medicines. The present study was designed to investigate the anti-inflammatory effect of the methanolic extract of Marrubiumvulgare, a popular traditional medicinal herb, on isoproterenol-induced myocardial infarction (MI) in rat model. METHODS: Male Wistar rats were assigned to 6 groups of control, sham, isoproterenol, and treatment with 10, 20, and 40 mg/kg/12h of the extract given orally concurrent with MI induction. A subcutaneous injection of isoproterenol (100 mg/kg/day) for two consecutive days was used to induce MI. Then, histopathological changes and inflammatory markers were evaluated. RESULTS: Isoproterenol injection increased inflammatory response, as shown by a significant increase in peripheral neutrophil count, myocardial myeloperoxidase (MPO) activity and serum levels of creatinine kinase-MB (CK-MB) and TNF-α (p<0.001). In the groups treated with 10, 20 and 40 mg/kg of M.vulgare extract serum CK-MB was subsided by 55.4%, 52.2% and 69%, respectively. Also treatment with the extract (40 mg/kg) significantly reduced (p<0.001) MPO activity in MI group. The levels of TNF-α was also considerably declined in the serums of MI group (p<0.001). In addition, peripheral neutrophil count, was significantly lowered by all doses of the extract (p<0.001). Interstitial fibrosis significantly was attenuated in treated groups compared with control MI group. CONCLUSION: The results of study demonstrate that the M. vulgare extract has strong protective effects against isoproterenol-induced myocardial infarction and it seems possible that this protection is due to its anti-inflammatory effects.

Hypoglycemic Activity of Fumaria parviflora in Streptozotocin-Induced Diabetic Rats.

PURPOSE: Fumaria parviflora Lam (Fumariaceae) has been used in traditional medicine in the treatment of several diseases such as diabetes. The present work was designed to evaluate the hypoglycaemic effects of methanolic extract (ME) of F. parviflora in normal and streptozotocin-induced diabetic rats. METHODS: The rats used were allocated in six (I, II, III, IV, V and VI) experimental groups (n=5). Group I rats served as 'normal control' animals received distilled water and group II rats served as 'diabetic control' animals. Diabetes mellitus was induced in groups II, V and VI rats by intraperitoneal single injection of streptozotocin (STZ, 55 mg kg-1). Group V and VI rats were addi-tionally treated with ME (150 mg kg-1 day-1 and 250 mg kg-1 day-1, i.p. respectively) 24 hour post STZ injection, for seven consecutive days. Groups III and IV rats received only ME 150 mg kg-1 day-1 and 250 mg kg-1 day-1, i.p. respectively for seven days. The levels of blood glucose were determined using a Glucometer. RESULTS: Administra-tion of F. parviflora extract showed a potent glucose lowering effect only on streptozo-tocin (STZ) induced diabetic rats below 100 mg/dl (P<0.001). However, no significant differences in the blood glucose levels were recorded between diabetic rats received 125 or 250 mg/kg of plant extracts. CONCLUSION: The findings of the study indicated that F. parviflora has significant hypoglycemic effect on STZ-induced diabetic rats with no effects on blood glucose levels of normal rats.

Antiproliferative activity of Marrubium persicum extract in the MCF-7 human breast cancer cell line.

AIM: Developing antitumor drugs from natural products is receiving increasing interest worldwide due to limitations and side effects of therapy strategies for the second leading cause of disease related mortality, cancer. METHODS: The antiproliferative activity of a methanolic extract from the aerial parts of Marrubium persicum extract was assessed with the MCF-7 breast cancer cell line using the MTT test for cell viability and cytotoxicity indices. In addition, antioxidant properties of the extract were evaluated by measuring its ability to scavenge free DPPH radicals. Moreover, the total phenolic and flavonoid content of the extract was determined based on Folin-Ciocalteu and colorimetric aluminum chloride methods. RESULTS: The findings of the study for the antiproliferative activity of the methanolic extract of M. persicum showed that growth of MCF-7 cells was inhibited by the extract in a dose and time dependent manner, where a gradual increase of cytotoxicity effect has been achieved setting out on 200 µg/mL concentration of the plant extract. The antioxidant assay revealed that the extract was a strong scavenger of DPPH radicals with an RC50 value of 52 µg/mL. The total phenolic and flavonoids content of the plant extract was 409.3 mg gallic acid equivalent and 168.9 mg quercetin equivalent per 100g of dry plant material. CONCLUSION: Overall, M. persicum possesses potential antiproliferative and antioxidant activities on the malignant MCF-7 cell line that could be attributed to the high content of phenolics and flavonoids, and therefore warrants further exploration.

Anti-oxidative role of quercetin derived from Allium cepa on aldehyde oxidase (OX-LDL) and hepatocytes apoptosis in streptozotocin-induced diabetic rat.

OBJECTIVE: To study the role of Quercetin in streptozotocin-induced diabetes in rats. METHODS: Wistar male rat (n=40) were allocated into three groups, control group (n=10) and Quercetin (QR) group received 15 mg/kg (IP) QR, (n=10), and diabetic group that received 55 mg/kg (IP) streptozotocin (STZ) (n=20) which was subdivided to two groups of 10; STZ group and treatment group. Treatment group received 55 mg/kg (IP) STZ plus 15 mg/kg QR, daily for 4 weeks, respectively; however, the control group just received an equal volume of distilled water daily (IP). Diabetes was induced by a single (IP) injection of streptozotocin (55 mg/kg). Animals were kept in standard condition. Twenty-eight days after inducing diabetic, 5 mL blood were collected for TAC, MDA and Ox-LDL levels and liver tissues of rat in whole groups were removed then prepared for apoptosis analysis by Tunel method. RESULTS: Apoptotic cells significantly decreased in group that has received 15 mg/kg (IP) Quercetin (P<0.05) in comparison to experimental groups (P<0.05). CONCLUSIONS: Since in our study 15 mg/kg (IP) Quercetin have significantly Preventive effect on liver cells damages by reducing number of apoptotic cells in Liver, so it seems that using it can be effective for treatment in diabetic rat.

Protective effect of ginger on gentamicin-induced apoptosis in testis of rats.

PURPOSE: Ginger, the rhizome of Zingiber officinale, is one of the most widely used spices for various foods and as an herbal medicine in Asian countries. It has been shown that ginger has antioxidant power. Gentamicin is an aminoglycoside antibiotic with a very broad spectrum against microbial pathogens, especially the gram-negative. Many studies revealed that gentamicin induces an oxidative stress-status in the testis by increasing free radical formation and lipid peroxidation. The present study was designed to investigate on the effects of Ginger as a natural anioxidant on testis apoptosis after treatment with gentamicin in rats. METHODS: In order to study the recovery effects of ginger on testis apoptosis after treatment with gentamicin 40 adult Wistar male rats were selected and randomly divided into four groups. Normal salin control (group I) (n=10), gentamicin control (group II), ginger control (group III) and gentamicin + ginger (group IV) each 10 rats. There was observation of negative effect of Gentamicin on testis histology in rats. RESULTS: The results revealed that there was a significant increase in apoptosis in group III when compared with other groups (P<0.05).However, ginger could decrease apoptosis in group IV that received 100mg/kg/rat of Ginger. CONCLUSION: Regarding the results, it is recommended that administration of ginger with gentamicin might be beneficial in men who receive gentamicin to treat infections.

Beneficial effects of quercetin on sperm parameters in streptozotocin-induced diabetic male rats.

Quercetin (QR) is a strong antioxidant and has been shown to reduce oxidative stress in the long-term treatment of streptozotocin (STZ)-induced diabetes in animal models. Antioxidants have significant effects on spermatogenesis, sperm biology and oxidative stress, and changes in antioxidant capacity are considered to be involved in the pathogenesis of chronic diabetes mellitus. The present study aims to examine the influence of QR on spermatogenesis in STZ-induced diabetes in male Wistar rats. Animals (n = 50) were allocated into five groups: Group 1: Control rats given 0.5 ml of 20% glycerol in 0.9% normal saline. Group 2: Control rats given buffer (pH4.0).Group 3: diabetic controls. Group 4: rats given QR 15 mg/kg/day (i.p.). Group 5: STZ + QR rats. Animals were kept in standard conditions. At the end of the experiment (28th day), blood samples were taken for determination of testosterone, total antioxidant capacity, and levels of malondialdehyde and oxidized low-density lipoprotein. All rats were euthanized, testes were dissected out and spermatozoa were collected from the epididymis for analysis. Sperm numbers, percentages of sperm viability and motility, and total serum testosterone increased significantly in QR-treated diabetic rats (P < 0.05) compared with control groups. In histopathology, degeneration and inflammation in testes cells associated with diabetes were improved and testes weights in the QR-treated diabetic group decreased significantly in comparison with controls (P < 0.05). We conclude that QR has significant beneficial effects on the sperm viability, motility, and serum total testosterone and could be effective for maintaining healthy sperm parameters and male reproductive function in diabetic rats.

The effect of total extract of Securigera securidaca L. seeds on serum lipid profiles, antioxidant status, and vascular function in hypercholesterolemic rats.

AIM OF THE STUDY: Seeds of Securigera securidaca are used for the treatment of disorders such as hyperlipidemia, diabetes, and epilepsy in Iranian folk medicine. The possible hypolipidemic and antioxidative effects of hydroalcoholic extract of S. securidaca seeds as well as the effect of the extract on vascular reactivity were investigated in hypercholesterolemic rats. MATERIALS AND METHODS: High-fat fed wistar rats received orally different doses of the extract for 20 days. At the end of the experiment vein blood and liver were collected to measure the lipid profile, lipid peroxidation, and antioxidative enzyme activities. The thoracic aorta was excised and used for isolated vessel preparation and histological study. RESULTS: The extract produced significant reductions (p<0.05) in the level of low-density lipoprotein (LDL) and triglyceride with concomitant reduction in lipid deposition in the liver. The extract also suppressed markedly (p<0.001) the hypercholesterolemia-induced elevation of malondialdehyde levels both in serum and liver. In hypercholesterolemic rats the endothelium-dependent vasodilatation was improved significantly (p<0.05) by 100mg/kg/day of the extract. However, in histological study no atherosclerotic lesion was observed. CONCLUSION: These results suggest that S. securidaca seed in addition to decrease lipid levels and peroxidation, is able to improve vascular endothelium-dependent relaxation in hypercholesterolemia.

Antinociceptive properties of extracts and two flavonoids isolated from leaves of Danae racemosa.

The antinociceptive properties of the hydro-methanolic extract (HME) and two flavonoids isolated from Danae racemosa have been investigated in several nociceptive rat models. The HME from D. racemosa (100-400 mgkg(-1), i.p.) produced significant dose-related inhibition of acetic acid-induced abdominal constriction. In the same dose range, the HME produced dose-related inhibition in both phases of a formalin-test. Treatment of animals with naloxone (5 mgkg(-1), i.p.) completely reversed the antinociceptive effect caused by morphine (5 mgkg(-1), s.c.) and the HME (200 mgkg(-1), i.p.) when assessed against the first phase of the formalin-test, but this effect was less significant for the HME in the second phase. Furthermore, when assessed via a hot-plate test, the HME (100-400 mgkg(-1), i.p.) caused a significant increase in response latency. The HME, given daily for to 7 consecutive days, develop tolerance, but did not induce cross-tolerance to morphine. These data demonstrate that the HME elicites pronounced antinociception against several pain models. The actions of the HME involve, at least in part, an interaction with the opioid system, but does not seem to be related with non-specific peripheral or central depressant actions. Finally, the active principle(s) responsible for the antinociceptive action of D. racemosa is likely to be partially related to the presence of quercetin and kaempferol.