The effects of methanolic extract of Glycyrrhiza glabra on the prevention and treatment of bleomycin-induced pulmonary fibrosis in rat: experimental study.

Idiopathic pulmonary fibrosis (IPF) is a chronic lung disease characterized by dyspnea and progressive loss of lung function. In this study, the preventive and therapeutic effects of methanolic extract of Glycyrrhiza glabra on pulmonary fibrosis were investigated. Pulmonary fibrosis was induced by administration of bleomycin (BLM) into the left lung of rats. Methyl-prednisolone (M-pred, 4 mg/kg) and methanolic extract of G. glabra (500 mg/kg) were injected intraperitoneally from the 1st to 14th days in the preventive group and from the 14th to 28th days in the therapeutic group once every day. Pulmonary inflammatory and fibrotic indices were evaluated by hematoxylin and eosin (H&E) and Masson's trichrome, respectively. The level of hydroxyproline as an index of pulmonary fibrosis and malondialdehyde (MDA) as an oxidative stress biomarker and catalase were measured by the related ELISA Kits. Pulmonary inflammatory and fibrotic indices in the G. glabra and M-pred groups significantly reduced compared with BLM group. G. glabra decreased the level of hydroxyproline in pulmonary tissue similar to M-pred. MDA reduced in G. glabra and M-pred groups compared with BLM group. The activity of catalase increased in the G. glabra preventive group. According to the results, G. glabra prevented and treated pulmonary fibrosis and inflammation in rats. Therefore, G. glabra may be suggested for the prevention and treatment of pulmonary fibrosis and inflammation.

Cytotoxicity evaluation of curcumin-loaded affibody-decorated liposomes against breast cancerous cell lines.

Curcumin is known as an effective anticancer herbal medicine but unfortunately, its bioavailability is poor which necessitate efforts for developing more efficient and specific delivery systems. Human epidermal growth factor receptor 2 (HER 2) due to its overexpression in various types of cancers, is demonstrated to be a good candidate as a target for anticancer therapy. In this study, cytotoxicity of curcumin encapsulated in ZHER2:342 Affibody-decorated liposome was investigated against SKBR3 and MCF-7 cancerous cell lines. Curcumin-containing liposomes were prepared from soybeans lecetin and cholesterol by thin-film hydration method. Affibody ZHER2:342 molecules via C-terminal cysteine residue were conjugated covalently to the prepared liposomes. Particle size analysis was performed using atomic force microscopy (AFM) and dynamic light scattering (DLS). Curcumin loading was measured using UV-Vis spectrophotometry and cytotoxic activity of curcumin formulations against cancerous cell lines was investigated by MTT assay. Induction of apoptosis was investigated using flow cytometry through Annexin V staining. Particle analysis showed the formation of spherical liposomes with a mean diameter of about 150 nm. Cytotoxic activity of curcumin was improved by its encapsulation in both liposomes and affibody-decorated liposomes. The Annexin V staining indicated the induction of apoptosis by affibody-decorated liposomes in both MCF-7 and SKBR3 cells. Decoration of curcumin-loaded liposomes with affibody ZHER2:342 may improve curcumin apoptotic function independently of HER2 expression level.

Nigella sativa L., Supplementary Plant with Anticholinesterase Effect for Cognition Problems: A Kinetic Study.

BACKGROUND: The average lifespan and the aging population are rising worldwide. So Neurodegenerative Disease (ND) will be one of the most common challenges associated with this population and would be more prevalent in future. The use of Acetylcholinesterase (AChE) inhibitors is one of the most important strategies for memory impairment. Medicinal plants are the most known natural source for accessing the new therapeutic agents. OBJECTIVE: In this work, we aimed to study in vitro anticholinesterase effect of different concentrations (10, 100, 250, 500, 750 and 1000 µg/ml) of total extract of N. sativa (NTE) and its separated fractions and to study the kinetic of AChE enzyme in the presence of two concentrations of NTE (10 and 100 µg/ml). METHODS: Maceration method was used for NTE preparation and different fractions of Petroleum Ether (PTE), Chloroform (CHF) and Methanol (MF). NTE, fractions and the main component of the plant, Thymoquinone (TQ), were assayed for AChE inhibition, using Ellman's method. Kinetic study of the AChE enzyme was studied in the presence of NTE at 10 and 100 µg/ml using Linweaver- Burk plot too. RESULTS: NTE and all the separated fractions inhibited AChE enzyme in a concentration-dependent manner. The greatest inhibition was shown by CHF and PEF fractions (86.97% and 79.99% at 1000 µg/ml, respectively). With less intensity, NTE, TQ and MF exhibited 76.32%, 68.98 % and 48.39% enzyme inhibition at 1000µg/ml, respectively. The least IC50 value was due to CHF fraction in AChE inhibition (98.28 ± 6.74 µg/ml). Kinetic profile exhibited the mixed mode of AChE inhibition by NTE. This indicates that a particular substance could not be responsible for AChE inhibition, and probably a collection of phytochemicals are involved in this process. CONCLUSIONS: N. sativa is a good candidate for seeking the new anticholinesterase agent and could be considered as a good supplement for the health of the elderly.

Zataria multiflora Boiss. extract improves spatial memory and learning capacity in scopolamine-induced amnesic rats.

OBJECTIVE: 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. MATERIAL AND METHODS: 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. RESULTS: 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. CONCLUSION: Our findings indicate that Zm has anti-amnesic effect and might improve memory deficit through anticholinesterase activity.

Oxidative stress in opium users after using lead-adulterated opium: The role of genetic polymorphism.

Use of lead-adulterated opium has become one of the major sources of lead poisoning in Iran. This study was designed to assess clinical effects and oxidative stress and its association with GSTM1, GSTT1, NQO1, and ALAD genes polymorphisms and blood lead level (BLL) in lead-adulterated opium users. The oxidative stress status in 192 opium users with lead poisoning symptoms measured and compared with 102 healthy individuals. Gluthatione S-transferase (GST)-M1 and -T1 genes deletion, NQO1 rs1800566, and δ-aminolevulinic acid dehydratase (ALAD) rs1800435 polymorphisms were determined using PCR and PCR-RFLP. The relation between the polymorphisms, BLL, and oxidative stress parameters were analysed using multivariate linear regressions. The common symptoms of lead toxicity were gastrointestinal and neurologic complications. Oxidative stress was significantly higher in opium addicts and lipid peroxidation significantly correlated with BLL. There was significant association between ALAD rs1800435 and BLL, and the BLL was significantly lower in the patients with ALAD 1-2 genotype. Use of lead-adulterated opium causes high frequency of lead toxicity symptoms, hematological and biochemical abnormalities, and oxidative stress which are associated with BLL. Route of opioid use and the polymorphism of rs1800435 in ALAD gene are the major determinants of BLL in lead-adulterated opium users.

Protective effect of standardized extract of Myrtus communis L. (myrtle) on experimentally bleomycin-induced pulmonary fibrosis: biochemical and histopathological study.

CONTEXT: Myrtle (Myrtus communis L) has been used widely in traditional medicine for different respiratory disorders. Idiopathic pulmonary fibrosis (IPF) is an inflammatory disease characterized by progressive loss of lung function with poor prognosis. The pathogenesis of disease has not been completely elucidated, but probably persistent epithelial damages are involved. OBJECTIVE: Evaluation of biochemical and histopathological effect of preventive and therapeutic doses of myrtle against bleomycin (BLM)-induced pulmonary fibrosis (PF) in animal model. MATERIALS AND METHODS: Methanolic extract of M. communis was prepared by maceration method. Total flavonoid content was determined and experimentally PF was induced in rat with intratracheal instillation of a single dose of BLM (5 mg/kg) only on day 0. Myrtle antifibrotic effect was evaluated as preventive (50 mg/kg/day, intraperitoneal (i.p.) injection, from day 0-13) and therapeutic agent (50 mg/kg, i.p., from day 14-27) in comparison with methyl prednisolone (M-pred) (4 mg/kg, i.p. for 14 days). RESULTS: Parenchymal inflammation and fibrotic changes significantly were reduced by myrtle and M-pred. Significant decrease in hydroxyproline content and lipid peroxidation were observed in animals receiving myrtle extract while catalase activity was increased by myrtle. Improvement in inflammation and fibrosis was observed in myrtle group especially in the early phase of fibrosis (preventive regime). DISCUSSION AND CONCLUSION: Myrtle extract effectively inhibited the inflammation and fibrosis of lung parenchyma in both preventive and therapeutic methods. This effect might be due to the reduction of tissue inflammation and inhibition of oxidative stress. More studies are being carried out to find main mechanisms and separation of active compounds.

Thymoquinone effectively alleviates lung fibrosis induced by paraquat herbicide through down-regulation of pro-fibrotic genes and inhibition of oxidative stress.

The potential preventive and therapeutic effects of thymoquinone (TQ) and its molecular mechanism were evaluated in paraquat (PQ)-induced pulmonary fibrosis in mice. TQ was administered orally at the doses of 20 and 40mg/kg during the course and after development of fibrosis. Pathological changes, expressions of genes involved in fibrogenesis, hydroxyproline (HP) and oxidative stress parameters were determined in the lung tissues. TQ dose-dependently recovered the pathological changes induced by PQ. TQ decreased hydroxyproline content, lipid peroxidation and restored the antioxidant enzymes to the normal values. In molecular level, expressions of TGF-ß1, α-SMA, collagen 1a1 and collagen 4a1 genes were also returned to the control level by TQ. This study indicated that TQ has the preventive and therapeutic potentials for the treatment of lung fibrosis by inhibition of oxidative stress and down-regulation of profibrotic genes.

Pharmacological safety evaluation of a traditional herbal medicine "Zereshk-e-Saghir" and assessment of its hepatoprotective effects on carbon tetrachloride induced hepatic damage in rats.

ETHNOPHARMACOLOGICAL RELEVANCE: "Zereshk-e-Saghir" (ZES), one of the traditional herbal medicines in old manuscripts of Persian hakims, has been used for the treatment of liver disorders. This current study is aimed to evaluate ZES effects on animal model to investigate its safety and hepatoprotective activity. MATERIALS AND METHODS: ZES was prepared according to a traditional method by blending aqueous extracts of Berberis vulgaris L., with fine particles of other plants including Rosa damascene Mill, Cichorium intybus L., Cucumis sativus L., Portulaca oleracea L., Rheum palmatum L., and Nardostachys jatamansi DC.. The lethality of ZES was determined in male NMRI mice. Acute organ toxicity of ZES (750 and 1500mg/kg for 15 days, orally) was evaluated by measuring the cell blood count, liver marker enzymes, creatinine, antioxidant status and histopathological examinations in rats. CCl4-induced liver toxicity was used to examine the hepatoprotective effects of the preparation. The rats were pretreated with 250, 500, 750 and 1500mg/kg ZES by gavage for 15 days. At day 16, the rats were intraperitoneally injected 1ml/kg CCl4 in olive oil. Forty-eight hours after CCl4 injection, the animals were sacrificed and their liver samples and blood were collected for determination of alanine aminotransferase, aspartate aminotransferase and alkaline phosphatase (ALT, AST, and ALP), histopathological examinations and antioxidant status. RESULTS: Treatment of the mice with a single dose of ZES up to 2g/kg did not cause mortality. Treatment of the rats with doses of 750 and 1500mg/kg for 15 days showed no significant hematotoxicity and hepatotoxicity. Treatment of the rats with ZES reduced the increased serum levels of ALT, AST, and ALP induced by CCl4 at the doses of 250, 500, and 750mg/kg. This was almost confirmed by histopathological examinations. Pretreatment with ZES also decreased lipid peroxidation and maintained the levels of glutathione and total antioxidant capacity. CONCLUSIONS: The present in vivo study revealed that the long term usage of ZES was safe for organs in laboratory animals. Meanwhile, prescribing the traditionally-recommended dose of ZES can be probably used against the liver injuries induced by xenobiotics. Further studies in other models of liver injuries are recommended for finding the exact hepatoprotective mechanism of ZES.

Chemical Composition, Anticonvulsant Activity, and Toxicity of Essential Oil and Methanolic Extract of Elettaria cardamomum.

Elettaria cardamomum is an aromatic spice (cardamom) native to the humid Asian areas, which contains some compounds with a potential anticonvulsant activity. Various pharmacological properties such as anti-inflammatory, analgesic, antioxidant, and antimicrobial effects have been related to this plant. This research was conducted to examine the probable protective impact of the essential oil and methanolic extract of E. cardamomum against chemically (pentylentetrazole)- and electrically (maximal electroshock)-induced seizures in mice. In addition, neurotoxicity, acute lethality, and phytochemistry of the essential oil and methanolic extract were estimated. The TLC method showed the presence of kaempferol, rutin, and quercetin in the extract, and the concentration of quercetin in the extract was 0.5 µg/mL. The major compounds in the essential oil were 1,8-cineole (45.6 %), α-terpinyl acetate (33.7 %), sabinene (3.8 %), 4-terpinen-4-ol (2.4 %), and myrcene (2.2 %), respectively. The extract and essential oil showed significant neurotoxicity in the rotarod test at the doses of 1.5 g/kg and 0.75 mL/kg, respectively. No mortalities were observed up to the doses of 2 g/kg and 0.75 mL/kg for the extract and essential oil. The essential oil was effective in both the pentylentetrazole and maximal electroshock models; however, the extract was only effective in the pentylentetrazole model. The study suggested that E. cardamomum methanolic extract had no significant lethality in mice. Both the essential oil and methanolic extract showed movement toxicity. Anticonvulsant effects of E. cardamomum were negligible against the seizures induced by pentylentetrazole and maximal electroshock.

Accelerated Burn Wound Closure in Mice with a New Formula Based on Traditional Medicine.

BACKGROUND: A combination of the oils of sesame, hemp, wild pistachio, and walnut has been used for treatment of skin disorders, including wound burns, in some parts of Kerman, Iran. Evaluation of this remedy in the form of a pharmaceutical formulation in animal models can pave the way for its future application in wound burn healing in humans. OBJECTIVES: This experimental study investigated the healing potential of a new formula (NF) based on folk medicine from Iran for the treatment of third degree burns in mice. The formula was a combination of the oils of four plants: sesame (Sesamum indicum L.), wild pistachio (Pistacia atlantica Desf.), hemp (Cannabis sativa L.), and walnut (Juglans regia L.). METHODS: Twenty-four mice were selected based on simple random sampling. Twenty-five percent of the total body surface area was burned by exposure to boiling water, according to the Walker-Mason method. NF and silver sulfadiazine (the positive control) were applied topically twice a day for 21 days. The burned area in the negative control group was left untreated. Epithelialization time and the percent of wound contraction were measured during the treatment period. The process of wound repairing was evaluated using histological (H and E and trichrome staining) and immunohistological (anti-pancytokeratin) methods. RESULTS: When compared to the controls, NF significantly improved wound contraction after day 10. Epithelialization time in the NF group was significantly faster than in the other groups (20 vs. 25.5 days) (P < 0.001). Histopathological and immunohistochemical findings confirmed the efficacy of the NF. CONCLUSIONS: A new therapeutic remedy was introduced for the treatment of burn wounds. Further clinical and molecular studies are suggested to determine the exact mechanism(s) involved in the burn wound healing effect of NF.

Anticholinesterase, antioxidant, and neuroprotective effects of Tripleurospermum disciforme and Dracocephalum multicaule.

BACKGROUND: Nowadays, owing to medicinal plants as a candidate to obtain promising new medicinal agents, there is a renewed interest in the use of these natural sources for drug development. OBJECTIVE: In the present study, we aimed to assess the anticholinesterase, antioxidant, and neuropotective effects of Tripleurospermum disciforme and Dracocephalum multicaule extracts. MATERIALS AND METHODS: Methanolic extract of the plants was prepared by maceration method. Anticholinesterase effect of different concentrations of the plants was studied by colorimetric method and antioxidant activity was evaluated using diphenypicrylhydrazil (DPPH) assay. Protective effect of the extracts against amyloid ß (Aß)-induced toxicity in PC12 cells was determined by MTT (3-(4,5-dimethyl thiazol-2-yl)-2,5-diphenyl tetrazolium bromide) method. RESULTS: Both T. disciforme and D. multicaule extracts could inhibit acetylcholinesterase (AChE) in a dose-dependent manner. The highest inhibition occurred at 5 µg/ml (71.18 ± 4.9 and 79.06 ± 3.1% inhibition respectively by T. disciforme and D. multicaule) in comparison to tacrine (86.37 ± 3.24%). The greatest DPPH inhibition of T. disciforme and D. multicaule was shown at 800 µg/ml (89.04 ± 3.9 and 78.5 ± 3.7%, respectively). None of tested extracts induced protection against ßA toxicity in PC12 cell. CONCLUSION: Although the results indicated anticholinesterase and antioxidant of the T. disciforme and D. multicaule, further specific studies and scientific validity are needed.

Anticonvulsant effect of the essential oil and methanolic extracts of Zataria multiflora Boiss.

UNLABELLED: Zataria multiflora is an Iranian endemic plant with a variety of uses. Anticonvulsant effect of the plant has not been studied yet. We aimed to evaluate the anticonvulsant effect of the essential oil and methanolic extract of the plant on chemical and electrically seizures. METHODS: Essential oil (EO) and methanolic extract (ME) were prepared by hydrodistillation and maceration method respectively and administered for evaluating the anticonvulsant activity against pentylenetetrazole (PTZ) and maximal electroshock (MES) convulsions in mice. Different doses of the EO and ME of Zataria multiflora were administered intraperitoneally (i.p), thirty minutes before induction of chemical (PTZ, 100 mg/kg) and electrical (50 mA, 50 Hz, 1 s duration) convulsions. Neurotoxicity was evaluated using rota-rod test. Mortality was determined after 24 hours for determining LD50. RESULTS: EO significantly increased the onset time of clonic seizures at doses of 0.2, 0.25 and 0.35ml/kg and significantly prevented tonic convulsions induced by PTZ at the same doses. No increase in onset time of tonic convulsions was observed. Methanolic extract of Z. multiflora was not effective in PTZ model. Neither EO nor ME was effective against MES convulsions; however both of them significantly reduced the time spent on rota-rod at doses of 0.6 ml/kg and 3 g/kg respectively. LD50 value of the EO was determined as 1.30 (1.0-1.5) ml/kg. No mortality was seen for the ME until dose of 5 g/kg. CONCLUSION: EO would be a good candidate for more investigations to determine its active constituent(s) and also its mechanism against PTZ-induced seizures.

Anticonvulsant activity and toxicity of essential oil and methanolic extract of Zhumeria majdae Rech, a unique Iranian plant in mice.

Drug-resistance and adverse effects of current drugs are the most obstacles in the treatment of epilepsy. In a plan for finding new natural anticonvulsant agents, we studied the anticonvulsant effects of essential oil (ZMEO) and methanolic extract (ZMME) of Zhumeria majdae in pentylene tetrazol (PTZ) and maximal electro-shock (MES) models in mice. Mice received different doses of ZMEO and ZMME, 30 min before induction of chemical and electrical convulsions. Neurotoxicity (movement toxicity and sedation) was evaluated using rota-rod test. The mortality was determined after 24 h following injection of different doses of the ZMEO and ZMME. The obtained results show that ZMEO dose-dependently protected mice from tonic convulsions induced by PTZ and MES with effective doses (ED(50)) of 0.26 (0.13-0.39) and 0.27 (0.17-0.37) ml/kg respectively. Toxic doses (TD(50)) in rota-rod test for ZMEO was 0.55 (0.42-0.70) ml/kg. ZMME at dose of 2 g/kg decreased tonic convulsions as much as 40 %. For ZMEO, TD(50) of 0.55 (0.45-0.69) ml/kg was obtained. ZMME significantly decreased the walking time in rota-rod test at dose of 2 g/kg. Lethal dose (LD(50)) of ZMEO was determined as 2.35 (1.98-2.65) ml/kg. ZMME showed about 34 % death of the animals at dose 5 g/kg. The essential oil of Z. majdae could be a good candidate for further anticonvulsive studies.

Anticonvulsant activity of the essential oil and methanolic extract of Bunium persicum (Boiss). B. Fedtsch.

ETHNOPHARMACOLOGICAL RELEVANCE: Bunium persicum is an endemic plant to Iran which its seeds have a long history of medicinal uses. AIM OF THE STUDY: This work aimed to study the anticonvulsant effect of the essential oil and methanolic extract of the plant. MATERIALS AND METHODS: The essential oil and methanolic extract of the plant were studied against pentylenetetrazole (PTZ) and maximal electroshock (MES) induced convulsions in mice in different doses. The neurotoxicity of the essential oil and methanolic extract was investigated using rotarod method. RESULTS: The essential oil and methanolic extract prolonged the onset of clonic and tonic seizures in PTZ. The tonic seizures were prevented by essential oil in both methods at dose of 1mL/kg and higher doses. The methanolic extract inhibited PTZ-convulsions at dose 3g/kg and was ineffective against MES induced convulsion. CONCLUSIONS: The essential oil of the plant might be useful to control absence and grand mal seizures at dose 1mL/kg. This activity might be due to its content of monoterpenes.

Antioxidant Activity of Iranian Echium amoenum Fisch & C.A. Mey Flower Decoction in Humans: A cross-sectional Before/After Clinical Trial.

Medicinal plants are recognized as sources of natural antioxidants that can protect from biological system oxidative stress. The present cross-sectional before/after clinical trial was carried out to investigate the antioxidant properties of the decoction of the flowers of Echium amoenum Fisch & C.A. Mey in humans. A group of 38 healthy subjects was invited to use the E. amoenum (7 mg kg(-1)) twice daily for 14 days. Blood samples before and after entering the study were measured for lipid peroxidation level (LPO), total antioxidant capacity (TAC) and total thiol (SH) molecules. A significant reduction of blood LPO (24.65 +/- 11.3 versus 19.05 +/- 9.7, P = 0.029) was observed after 14 days of E. amoenum consumption. Blood TAC (1.46 +/- 0.51 versus 1.70 +/- 0.36, P = 0.018) and total thiol molecules (0.49 +/- 0.11 versus 0.56 +/- 0.12, P = 0.001) increased after 14 days of E. amoenum consumption. In conclusion, this antioxidative stress potential of E. amoenum may be due to its bioactive antioxidant components, especially rosmarinic acid and flavonoids. In recent years the importance of oxidative stress in the pathophysiology of many human disorders has been confirmed, thus use of this plant as a dietary supplement is highly recommended.