Ethnopharmacological studies of medicinal plants in central Zagros, Lorestan Province, Iran.

ETHNOPHARMACOLOGICAL RELEVANCE: Lorestan Province (Iran) has a great diversity of habitats and plant diversity and the people living in this province have a good knowledge of herbal therapies. AIM OF THE STUDY: In this study, the authors aimed to identify and report the medicinal plants used in the folk medicine of Lorestan. MATERIAL AND METHODS: The indigenous medicinal information was collected with a semi-structured open-ended questionnaire, interviews, and personal observations. The relative importance of the species were estimated through frequency of citations (FC). RESULTS: and discussion: A total of 555 plants belonging to 91 families were identified. The plants belong to Asteraceae with 62 species followed by Lamiaceae (56), Apiaceae (44), Fabaceae (41), and Brassicaceae (31). The plants were mostly used as a decoction or eaten raw (32%). Leaves (22%) were the most utilized plant parts followed by shoots (16%), and seeds (13%). Thymus daenensis, Thymus eriocalyx, Mentha longifolia, Mentha spicata, Mentha piperitha, Alium sativum, Quercus infectoria, Quercus persica, Ziziphora clinopodioides, and Malus domestica had the highest FC values. Most of the plants were used for gastrointestinal, respiratory, and skin problems. CONCLUSION: Lorestan is a region rich in medicinal plants. The local knowledge of herbal therapies can be used as complementary medicine, particularly by people in remote areas and as a guide for the future pharmacological and phytochemical studies. While the known medicinal plants can be harvested or cultivated for medical and pharmaceutical purposes, the less known plants with high FC scores can be investigated phytochemically and pharmacologically in the future studies.

Inhibition of protein glycation by essential oils of branchlets and fruits of Juniperus communis subsp. hemisphaerica.

Oxidative stress and protein glycation play pivotal roles in the pathophysiology of diabetes mellitus and its vascular complications. The present study aimed to investigate the anti-glycation properties of essential oils obtained from different parts of Juniperus communis subsp. hemisphaerica. The branchlets of male tree (BMT) and branchlets of female (BFT) tree, and fruits of J. communis subsp. hemisphaerica were extracted using steam distillation method. The oils were phytochemically analyzed using gas chromatography-mass spectrometry. Anti-glycation properties were evaluated using hemoglobin and insulin glycation assays. Overall, 18 volatile components were identified in the J. communis subsp. hemisphaerica oils, amounting to 82.1%, 100.0% and 96.4% of the BMT, BFT and fruit oils, respectively. Promising inhibitory activity was observed from all concentrations of the tested oils in the hemoglobin and insulin glycation assays. The inhibitory activities peaked to 89.9% (BFT oil; 200 µg mL(-1)) and 81.0% (BFT oil; 600 µg mL(-1)) in the hemoglobin and insulin glycation assays, respectively. The evidence from this study suggests that essential oils obtained from the fruits and branchlets of J. communis subsp. hemisphaerica possess anti-glycation properties. These activities may find implication for the prevention and treatment of diabetic complications.

Chemical composition and antiprolifrative activity of Artemisia persica Boiss. and Artemisia turcomanica Gand. essential oils.

Essential oils obtained from aerial parts of Artemisia persica and Artemisia turcomanica were analyzed by GC/MS. While 28 components representing 91.01 % of A. persica were identified, the identity of 50 components, constituting 81.93 % of the total oil, was confirmed in A. turcomanica. ß-thujone was the main compound (75.23%) in A. persica while the major identified phytochemicals in A. turcomanica were 1,8-cineol (19.23%), camphor (15.55%) and filifolone (15.53%). Both of the essential oils were predominantly made up of monoterpenes. Time- and dose-dependent cytotoxic effects of A. persica and A. turcomanica on MCF-7 cell line evaluated by MTT assay at 24, 48 and 72 h, showed that the highest cytotoxic effect of A. persica and A. turcomanica were appeared at 72 h incubation. At that incubation period, CI50 of A. persica was found to be 0.15 µg/ml, while that of A. turcomanica was 0.1 µg/ml. Thus, cytotoxicity of A. turcomanica was slightly higher than A. persica which could be attributed to the higher content of sesquiterpene present in A. turcomanica. As a conclusion, these volatile oils could have chemotherapeutic potentials.

Antioxidant activities of Juniperus foetidissima essential oils against several oxidative systems

The present study aimed to evaluate the antioxidant activity of essential oils obtained from branchlets of male and female trees as well as fruits of Juniperus foetidissima Willd., Cupressaceae, from Iran. For this purpose, essential oils of J. foetidissima were phytochemically analyzed and different concentrations of them were tested in five oxidative systems: 1) low-density lipoprotein oxidation; 2) linoleic acid peroxidation; 3) red blood cell hemolysis; 4) hemoglobin glycation; and 5) insulin glycation assays. In all employed systems, antioxidant effects were observed from the three tested oils though in varying degrees. The most promising activities of the oils were observed against hemoglobin and insulin glycation. Antioxidant activities of the oils did not appear to be dose-dependent. In addition, no consistent superiority in antioxidant effects was observed from a single oil in different assays. In view of the current results, J. foetidissima branchlet and fruit oils could be regarded as effective natural products with anti-glycation activity.

Molecular characterization and RAPD analysis of Juniperus species from Iran.

The genus Juniperus L. (Cupressaceae), an aromatic evergreen plant, consists of up to 68 species around the world. We classified five species of Juniperus found in Iran using molecular markers to provide a means for molecular identification of Iranian species. Plants were collected (three samples of each species) from two different provinces of Iran (Golestan and East Azarbayejan). The DNA was extracted from the leaves using a Qiagen Dneasy Plant Mini Kit. Amplification was performed using 18 ten-mer RAPD primers. Genetic distances were estimated based on 187 RAPD bands to construct a dendrogram by means of unweighted pair group method of arithmetic means. It was found that J. communis and J. oblonga were differentiated from the other species. Genetic distance values ranged from 0.19 (J. communis and J. oblonga) to 0.68 (J. communis and J. excelsa). Juniperus foetidissima was found to be most similar to J. sabina. Juniperus excelsa subspecies excelsa and J. excelsa subspecies polycarpos formed a distinct group.

The potential inhibitory effect of cuminum cyminum, ziziphora clinopodioides and nigella sativa essential oils on the growth of Aspergillus fumigatus and Aspergillus flavus

The goals of this study were to evaluate the effectiveness of Cuminum cyminum, Ziziphora clinopodioides and Nigella sativa essential oils to inhibit the growth of Aspergillus fumigatus and A. flavus and to evoke ultrastructural changes. The fungi were cultured into RPMI 1640 media in the presence of oils at concentrations of 8, 6, 5, 4, 3, 2, 1.5, 1.25, 1, 0.75 and 0.5 mg/ml in broth microdilution and 2, 1.5, 1 and 0.5 mg/ml in broth macrodilution methods with shaking for 48 h at 28ºC. Conidial and mycelial samples exposed to 0.25, 0.5, 1, 1.5 and 2 mg essential oils/ml for 5 days in 2 percent yeast extract granulated plus 15 percent Saccharose media were processed for transmission electron microscopy (TEM). Based on broth dilution methods, C. cyminum and to a lesser extent Z. clinopodioides oils exhibited the strongest activity against A. fumigatus and A. flavus with MIC90 ranging from 0.25 to 1.5 mg/ml, while the oil from N. sativa exhibited relatively moderate activity against two above fungi with MIC90 ranging from 1.5 to 2 mg/ml. The main changes observed by TEM were in the cell wall, plasma membrane and membranous organelles; in particular, in the nuclei and mitochondria. These modifications in fungal structure were associated with the interference of the essential oils with the enzymes responsible for cell wall synthesis, which disturbed normal growth. Moreover, the essential oils caused high vacuolation of the cytoplasm, detachment of fibrillar layer of cell wall, plasma membrane disruption and disorganization of the nuclear and mitochondrial structures. Aspergillus fumigatus and A. flavus growth inhibition induced by these oils were found to be well-correlated with subsequent morphological changes of the fungi exposed to different fungistatic concentrations of the oils. Our results show the anti-Aspergillus activities of C. cyminum, Z. clinopodioides and N. sativa essential oils, which strengthens the potential use of these substances as anti-mould in the future.

The potential inhibitory effect of cuminum cyminum, ziziphora clinopodioides and nigella sativa essential oils on the growth of Aspergillus fumigatus and Aspergillus.

The goals of this study were to evaluate the effectiveness of Cuminum cyminum, Ziziphora clinopodioides and Nigella sativa essential oils to inhibit the growth of Aspergillus fumigatus and A. flavus and to evoke ultrastructural changes. The fungi were cultured into RPMI 1640 media in the presence of oils at concentrations of 8, 6, 5, 4, 3, 2, 1.5, 1.25, 1, 0.75 and 0.5 mg/ml in broth microdilution and 2, 1.5, 1 and 0.5 mg/ml in broth macrodilution methods with shaking for 48 h at 28(o)C. Conidial and mycelial samples exposed to 0.25, 0.5, 1, 1.5 and 2 mg essential oils/ml for 5 days in 2% yeast extract granulated plus 15% Saccharose media were processed for transmission electron microscopy (TEM). Based on broth dilution methods, C. cyminum and to a lesser extent Z. clinopodioides oils exhibited the strongest activity against A. fumigatus and A. flavus with MIC90 ranging from 0.25 to 1.5 mg/ml, while the oil from N. sativa exhibited relatively moderate activity against two above fungi with MIC90 ranging from 1.5 to 2 mg/ml. The main changes observed by TEM were in the cell wall, plasma membrane and membranous organelles; in particular, in the nuclei and mitochondria. These modifications in fungal structure were associated with the interference of the essential oils with the enzymes responsible for cell wall synthesis, which disturbed normal growth. Moreover, the essential oils caused high vacuolation of the cytoplasm, detachment of fibrillar layer of cell wall, plasma membrane disruption and disorganization of the nuclear and mitochondrial structures. Aspergillus fumigatus and A. flavus growth inhibition induced by these oils were found to be well-correlated with subsequent morphological changes of the fungi exposed to different fungistatic concentrations of the oils. Our results show the anti-Aspergillus activities of C. cyminum, Z. clinopodioides and N. sativa essential oils, which strengthens the potential use of these substances as anti-mould in the future.

Growth-inhibitory effect of Scutellaria lindbergii in human cancer cell lines.

Scutellaria lindbergii (Lamiaceae) is Iranian species of Scutellaria. Cytotoxic properties of total methanol extract of S. lindbergii and its fractions were investigated on different cancer cell lines including AGS, HeLa, MCF-7, and PC12. Meanwhile the role of apoptosis was explored in this toxicity. Malignant and non-malignant cells were cultured in DMEM medium and incubated with different concentrations of plant extracts. Cell viability was quantitated by MTT assay. Apoptotic cells were determined using PI staining of DNA fragmentation by flow cytometry (sub-G1 peak). S. lindbergii inhibited the growth of malignant cells in a dose-dependent manner. Among solvent fractions of S. lindbergii, the methylene chloride fraction was found to be more toxic compared to other fractions. S. lindbergii-induced a sub-G1 peak in flow cytometry histogram of treated cells compared to control indicating apoptotic cell death is involved in S. lindbergii-induced toxicity. In conclusion, S. lindbergii exerts cytotoxic effects in different cancer cell lines in which apoptosis plays an important role. Thus S. lindbergii could be considered as a potential chemotherapeutic agent in cancer treatment.

Antioxidant activity of leaves and fruits of Iranian conifers.

Cupressus semipervirens var. horizontalis, Cupressus semipervirens var. semipervirens, Cupressus semipervirens cv. Cereifeormis, Juniperus communis subsp. hemisphaerica, Juniperus excelsa subsp. excelsa, Juniperus excelsa subsp. polycarpos, Juniperus foetidissima, Juniperus oblonga, Juniperus sabina, Platycladus orientalis and Taxus baccata are Iranian conifers. The antioxidant activity of leaves and fruits of these 11 different taxons were evaluated. The leaves of both male and female, and fruits of these plants were collected from different areas of the country. Methanol extract of leaves and fruits of these taxons were prepared. Antioxidant activity of each extracts was measured using two different tests of the ferric thiocyanate method and thiobarbituric acid. Results indicated that the methanol extracts of leaves, of male and female, and fruits of all these species (27 samples) possessed antioxidant activity when tested with both methods. The antioxidant activity was then compared with those of alpha-tocopherol (a natural antioxidant) and butylated hydroxytoluene (a synthetic antioxidant). Methanol extract of fruits of C. semipervirens cv. Cereifeormis showed the highest antioxidant activity while the methanol extract of leaves of C. semipervirens var. semipervirens possessed the lowest antioxidant activity. However, our finding showed that most of the tested extracts were showing strong antioxidant activity even higher than alpha-tocopherol.

Microvascular reconstruction of soft tissue and bone loss in war wounds.

Seventy-seven skin and bone defects caused by high velocity missiles have been repaired with various combinations of microvascular free transfer of skin flaps and vascularised bone grafts. We concluded that the free osteoseptocutaneous fibula transfer (1,4) is the best method of reconstruction if a long bone defect is associated with skin loss.