Acute and subchronic toxicological evaluation of Echinophora platyloba DC (Apiaceae) total extract in Wistar rats

OBJECTIVE: Echinophora platyloba DC is a widely used herbal medicine and food seasoning in Iran. It is claimed to exert antimicrobial, antifungal, and antispasmodic effects. Despite the prevalent use of this plant as a food and medicine, there are no reports on its possible toxic effects. To evaluate the safety of E. platyloba, we tested its acute and sub-chronic toxicity in male and female Wistar rats. METHODS: Rats were orally treated with four different single doses of E. platyloba total extract and screened for signs of toxicity two weeks after administration. In the sub-chronic toxicity study, E. platyloba was administered for 45 days. Mortality, clinical signs, body weight changes, hematological and biochemical parameters, gross findings, organ weights, and histological markers were monitored during the study. RESULTS: We found no mortality and no abnormality in clinical signs, body weight, or necropsy findings in any of the animals in the acute study. The results of the subchronic study showed no significant difference in hematological parameters in either sex. There was a significant increase in lactate dehydrogenase in the female groups. A significant increase in the relative lung weight of female rats was noted at 500 mg/kg. Histopathological examinations revealed intra-alveolar hemorrhage in the male rats (500 mg/kg). In the females, congestion of the alveolar capillaries (at 500 mg/kg) and liver bridging necrosis (at 200 mg/kg) were significantly increased. CONCLUSION: The no observed adverse effect level of E. platyloba was determined to be 200 and 50 mg/kg for male and female rats, respectively

Effects of prantschimgin and grandivitin from Ferulago macrocarpa on VEGF, MMP9, MMP2 and research of binding modes using computational methods

Ferulago macrocarpa contains prantschimgin and grandivitin from phenolic class. Studies have shown that phenolic compounds at physiological concentrations could inhibit two groups of gelatinase matrix metalloproteinases (MMP2, MMP9). Due to the high diversity of coumarin in the plant, the possibility ability of the compounds to inhibit plant enzymes seem to be mentioned. introduction: Ferulago macrocarpa of Apiaceae, native to the highlands of the West of Iran (Zagros Mountains) includes Prantschimgin and Grandivitin from phenolic class. researches have shown that phenolic compounds at physiological concentrations could prevent two groups of gelatinase matrix metalloproteinases (MMP2, MMP9). Because of the high variety of coumarin in the plant, the potency of the compounds to inhibit plant enzymes seem to be refered. Methods and Results: Acetone extract of the plant was provided and then winterized. Thereafter, Prantschimgin and Grandivitin were purified using normal phase column chromatography and preparative HPLC, and their structures were verified. After culturing the cells, at confluence step, Supernatants were collected at24 and 48h soup (broth supernatant containing medium containing 2% albumin, and glutamine and antibiotics and enzymes secreted by the different concentrations of active ingredients) and non-proliferation medium containing 2% albumin was amplified. The pure substances were applied on cell lines U87MG and Wehi for evaluation of VEGF, MMP-2 and 9 activities. In the computational processing, the structures have been docked in the active site of metalloproteinases9, and important interactions were detennined. then, ligand-protein complexes were subjected to molecular dynamics simulation in water, and thermodynamic attributes were calculated. (MMP9 code= 1L6J, MMP2 code= 1CK7). prantschimgin and grandivitin were purified fromF.macrocarpafruits. Regarding cytotoxicity results IC50 of Prantschimgin and Grandivitin in Wehi cell line were 521.63,232.66, and in U87MG cell line were575.58,322.0 lpg/ml, respectively. Biological experiment indicated significant changes in the amount and activity of matrix metalloproteinase and vascular endothelial growth factor. Conclusion: Two coumarins, prantschimgin and grandivitin with the potential inhibitory effects on the activity of MMP2,9 and anti-angiogenesis were purified from F. macrocarpa fruits. The application can be expected to have therapeutic efficacy in cancer cell lines U87MG and Weh

Evaluation of Sesquiterpenes from Ferula assa-foetida on inflamatory parameters and study of biding modes using computational methods

introduction: Ferula assafoetida is a source of sesquiterpenes [1]. According to an investigation, phenolic compounds at physiological concentration can inhibit inflammatory enzymes [2]. These enzymes digest the extracellular matrix and provide the conditions for activation and migration and proliferation of endothelial cells. Reported studies on medicinal plants for their inhibitory effect on MMP are very limited. Methods and Results: Acetone extract of plant was prepared and Sesquiterpenes were purified using HPLC preparative analyses and their structures were elucidated. After culturing the cell at confluence, cells were isolated and the supernatant was removed. The pure substances were applied on cell lines U87MG and Wehi activities. Besides the structure has been docked in the active site of metalloproteinase, and significant interactions were determined.Subsequently, ligand-protein complexes were subjected to molecular dynamics simulation in water and thermodynamic properties were calculated. In the phytochemistry field galbanic acid, mogoltadone, kellerin, polyanthin and polyanthininwere produced from F. assafoetida. The results of celluar toxicity study shows that IC50 of Galbanic acid, Mogoltadone and Polyanthin in Wehi cell line were 925.2703, 721.86, and 680.3 µg/ml in U87MG cell line were 952.193, 752.352, 678.742. Galbanic acid, mogoltadone, kellerin, polyanthin and polyanthininwere solated from F. assafoetida. The results of celluar toxicity study show that IC50 of Galbanic acid, Mogoltadone and Polyanthin in Wehi cell line were 925.2703, 721.86, and 680.3 µg/ml in U87MG cell line were 952.193, 752.352, 678.742 Conclusion: Investigation revealed that the coumarins have inhibitory effects on the content and activity of MMP 2.9 and showed anti-angiogenetic effect. So, they can be potentially effective in the treatment of cancer. Interactive and competitive binding between MMP-9 and Galbanic acid were studied with FT-IR, UV-Vis and fluorescence methods and MMP-9 structure was changed in these interactions

Evaluation of the Effect of Six Terpenoids and Phenolics from Echinophora Cinerea against Cisplatin-Induced Oxidative Stress and Apoptosis in PC12 Cell Line

Introduction:Echinophoracinerea is a plant from Apiaceae family and it is used as vegetable, yogurt and cheese seasoning and is used for gasteric ailments in ChaharMahalBakhtiari province. It is a rich source of antioxidant constituents, hence it canpotentially  have protective effects. So, its phytochemical investigation seems to be crucial. Methods and Results:Plant material was extracted. The latter extract was fractionated with VLC and further purified using reversed phase HPLC. The structures of pure compounds were elucidated using spectroscopic methods such as 1HNMR and mass. Cytotoxic effects of cisplatin alone and with other fractions were tested. The effects of isolated compounds against apoptosis induced by CIS were investigated through the measurement of mitochondrial membrane potential, Bax and Bcl2 and caspase-3 activation. We also assessed the oxidative stress by measuring reactive oxygen species. Six compounds (quercetin-3-O-β-D-glucopyranoside, Kaempferol glycoside, osthol, verbenone, isoimperatorin and echinophorin B) were purified and identified. Treatment of cells with QUE and OST before exposure to the CIS increased cell viability. These compounds protected the cells against CIS–induced cytotoxicity. In addition, pretreatment with QUE and OST decreased CIS- induced apoptosis through up-regulation of Bcl2, inhibition of caspase-3 activity and increasing of mitochondrial membrane potential. As well, OST decreased ROS generation. Conclusion:Given that flavonoids are the most important groups of phenolic compounds found in nature, and due to their antioxidant and antiapoptotic effect these could be considered as neuroprotective agent

Comparative Evaluation of Cytotoxic and Apoptogenic Effects of Several Coumarins on Human Cancer Cell Lines: Osthole Induces Apoptosis in p53-Deficient H1299 Cells

Natural products are excellent resources for finding lead structures for the development of chemotherapeutic agents. Coumarins are a class of natural compounds found in a variety of plants. In this study, we evaluated the cytotoxic potential of coumarins isolated from Prangos ferulacea (L.) Lindl. in PC3, SKNMC, and H1299 (p53 null) human carcinoma cell lines. Osthole proved to be an outstanding potent cytotoxic agent especially against PC3 cells. Isoimperatorin exhibited moderate inhibitory effect against SKNMC and PC3 cell lines. Oxypeucedanin and braylin did not display any cytotoxic activity. In the next set of experiments, the apoptotic potentials of osthole and isoimperatorin were investigated. Induction of apoptosis by isoimperatorin was accompanied by an increase in activation of caspase-3, -8, and -9 in SKNMC cells and caspase-3 and -9 in PC3 cells. Moreover, isoimperatorin induced apoptosis by upregulating Bax and Smac/DIABLO genes in PC3 and SKNMC cells. Osthole induced apoptosis by downregulating antiapoptotic Bcl-2 in only PC3 cells and upregulating the proapoptotic genes Bax and Smac/DIABLO in PC3, SKNMC, and H1299 cells. The effects of osthole on H1299 cells are important because the loss of p53 has been associated with poor clinical prognosis in cancer treatment

Comparative Evaluation of Cytotoxic and Apoptogenic Effects of Several Coumarins on Human Cancer Cell Lines: Osthole Induces Apoptosis in p53-Deficient H1299 Cells

Natural products are excellent resources for finding lead structures for the development of chemotherapeutic agents. Coumarins are a class of natural compounds found in a variety of plants. In this study, we evaluated the cytotoxic potential of coumarins isolated from Prangos ferulacea (L.) Lindl. in PC3, SKNMC, and H1299 (p53 null) human carcinoma cell lines. Osthole proved to be an outstanding potent cytotoxic agent especially against PC3 cells. Isoimperatorin exhibited moderate inhibitory effect against SKNMC and PC3 cell lines. Oxypeucedanin and braylin did not display any cytotoxic activity. In the next set of experiments, the apoptotic potentials of osthole and isoimperatorin were investigated. Induction of apoptosis by isoimperatorin was accompanied by an increase in activation of caspase-3, -8, and -9 in SKNMC cells and caspase-3 and -9 in PC3 cells. Moreover, isoimperatorin induced apoptosis by upregulating Bax and Smac/DIABLO genes in PC3 and SKNMC cells. Osthole induced apoptosis by downregulating antiapoptotic Bcl-2 in only PC3 cells and upregulating the proapoptotic genes Bax and Smac/DIABLO in PC3, SKNMC, and H1299 cells. The effects of osthole on H1299 cells are important because the loss of p53 has been associated with poor clinical prognosis in cancer treatment

The Antimicrobial Activity of Different Extracts from Echinophora platyloba DC.

Background and Aim: Echinophora platyloba DC (Apiaceae) is a plant endemic to Kurdistan province of Iran. Aerial part of the plant is regarded as the source of natural antimicrobial agent. Materials and Methods: The aerial part of E. platyloba was extracted with hexane, dichloromethane, acetone EtOH and EtOH: H2O respectively. The extracts were individually tested against three gram-negative (Escherichia coli, Shigella flexneri, Acinetobacter baumannii) and two gram-positive bacteria (Staphylococcus aureus, Enterococcus faecalis) via agar dilution methods. Results: The best inhibitory effect was observed in ethanolic extract, which had a remarkable inhibitory effect on all bacteria especially on S. aureus. The results also indicated that dichloromethane extract showed the weakest inhibitory effect on all types of bacteria. Meanwhile, acetone extract exhibited a relatively mild inhibitory effect. Conclusion: The antimicrobial effects of E. platyloba confirmed its potential for folk use. More comprehensive investigations are recommended concerning the significant antibacterial activity of the ethanol extract of E. platyloba to determine the exact compounds responsible for observed biological activities

Protective effect of bioactive compounds from Echinophora cinerea against cisplatin-induced oxidative stress and apoptosis in the PC12 cell line

Objective(s): The present study aims to evaluate the protective effect of the compounds isolated from Echinophora cinerea (E. cinerea) against oxidative stress and apoptosis induced by cisplatin (CIS) in PC12 cells. Materials and Methods: Six compounds were isolated as quercetrin-3-O-β-D-glucopyranoside (QUE), osthol (OST), verbenone-5-O-β-D-glycopyranoside (VER), Isoimperatorin (ISO), kaempferol-3-O-β-D-glucopyranoside (KAM), and echinophorin B (ECH). For this study, we used MTT reduction assay for detection of protective effects of isolated compounds on CIS-induced cytotoxicity in PC12 cells. The effects of isolated compounds against apoptosis induced by CIS were investigated through the measurement of mitochondrial membrane potential (MMP), Bax and Bcl2 mRNA expression, and caspase-3 activation. We also assessed oxidative stress by measuring reactive oxygen species (ROS) generation with 2′, 7′-dichlorofluorescein diacetate (DCFH-DA). Results: Treatment of cells with QUE and OST before exposure to the CIS increased cell viability, i.e., these compounds protected the cells against CIS -induced cytotoxicity. In addition, pre-treatment with QUE and OST decreased CIS-induced apoptosis through up-regulation of Bcl-2, inhibition of caspase-3 activity, and mitochondrial membrane potential (MMP) increase. OST decreased ROS generation induced by CIS, as well. Conclusion: Our in vitro experiment showed that QUE and OST are apoptotic inhibitors that effectively block CIS-induced neurotoxicity predicting their therapeutic potential in the prevention of chemotherapy-induced neurotoxicity

Osthole Attenuates Doxorubicin-Induced Apoptosis in PC12 Cells through Inhibition of Mitochondrial Dysfunction and ROS Production

Doxorubicin (DOX) is a potent, broad-spectrum chemotherapeutic drug used for treatment of several types of cancers. Despite its effectiveness, it has a wide range of toxic side effects, many of which most likely result from its inherent prooxidant activity. It has been reported that DOX has toxic effects on normal tissues, including brain tissue. In the current study, we investigated the protective effect of osthole isolated from Prangos ferulacea (L.) Lindl. on oxidative stress and apoptosis induced by DOX in PC12 as a neuronal model cell line. PC12 cells were pretreated with osthole 2 h after treatment with different concentrations of DOX. 24 h later, the cell viability, mitochondrial membrane potential (MMP), the activity of caspase-3, the expression ratio of Bax/Bcl-2, and the generation of intracellular ROS were detected. We found that pretreatment with osthole on PC12 cells significantly reduced the loss of cell viability, the activity of caspase-3, the increase in Bax/Bcl-2 ratio, and the generation of intracellular ROS induced by DOX. Moreover, pretreatment with osthole led to an increase in MMP in PC12 cells. In conclusion, our results indicated that pretreatment with nontoxic concentrations of osthole protected PC12 cells from DOX-mediated apoptosis by inhibition of ROS production

The hydrogen sulfide releasing molecule acetyl deacylasadisulfide inhibits metastatic melanoma

Melanoma is the most common form of skin cancer. Given its high mortality, the interest in the search of preventive measures, such as dietary factors, is growing significantly. In this study we tested, in vitro and in vivo, the potential anti-cancer effect of the acetyl deacylasadisulfide (ADA), a vinyl disulfide compound, isolated and purified from asafoetida a foul-smelling oleo gum-resin of dietary and medicinal relevance. ADA markedly suppressed proliferation of human melanoma cell lines by inducing apoptosis. Moreover, treatment of melanoma cells with ADA reduced nuclear translocation and activation of NF-κB, decreased the expression of the anti-apoptotic proteins c-FLIP, XIAP, and Bcl-2 and inhibited the phosphorylation and activation of both AKT and ERK proteins, two of the most frequently deregulated pathways in melanoma. Finally, the results obtained in vitro were substantiated by the findings that ADA significantly and dose-dependently reduced lung metastatic foci formation in C57BL/6 mice. In conclusion, our findings suggest that ADA significantly inhibits melanoma progression in vivo and could represent an important lead compound for the development of new anti-metastatic agents