The role of quercetin in the formation of titanium dioxide nanoparticles for nanomedical applications.

The current work aimed to synthesize and characterize titanium dioxide nanoparticles (TiO2NPs) using quercetin (QE) and evaluate their biological activities, i.e., anti-hemolytic, anti-inflammatory, and cytotoxicity effects. The crystallographic phase and morphology of biosynthesized QE-TiO2NPs were characterized by XRD (X-Ray Diffraction) and TEM/FE-SEM (Transmission/Field-Emission Scanning Electron Microscopy) micrographs. Functional groups involved in the synthesis process were determined by FTIR spectroscopy (Fourier Transform-Infrared Spectroscopy). Based on the characterization results, selected QE-TiO2NPs showed a rutile phase, spherical shape, and a size range of 7.3-39 nm. The QE-TiO2NPs did not show a hemolytic effect. They indicated 95.3% red blood cells (RBCs) membrane stabilization activity and 82.6% inhibition of bovine serum albumin (BSA) denaturation, similar to a standard drug, which proved their anti-inflammatory effects. The attained results from cytotoxicity studies revealed the toxic effects of QE-TiO2NPs with IC50 values below 100 and 50 µg/mL for human breast cancer cells of MCF-7 and melanoma cancer cells of A375, respectively. These NPs did not significantly affect normal skin fibroblast cells up to 50 µg/mL and only showed a 16% inhibition rate on the cell viability at 100 µg/mL. These NPs also induced excessive ROS generation. This work established the blood/biocompatibility and excellent nanomedical applications of biosynthesized QE-TiO2NPs.

Colchicine of Colchicum autumnale, A Traditional Anti-Inflammatory Medicine, Induces Apoptosis by Activation of Apoptotic Genes and Proteins Expression in Human Breast (MCF-7) and Mouse Breast (4T1) Cell Lines.

OBJECTIVE: Breast cancer is one of the major causes of mortality among women. Due to many side effects of the existing chemotherapeutic agents, the research of anti-cancer drugs, including natural products, is still a big challenge. Here, we investigated the effects of colchicine on apoptosis of two breast cancer cell lines ( human MCF-7 and mouse 4T1). MATERIALS AND METHODS: In this experimental study, we evaluated the apoptotic effects of colchicine on (MCF-7) and (4T1), as well as a human cancer-associated fibroblast cell line as a control group. Extraction and chromatographic techniques were applied to isolate colchicine from Colchicum autumnale L. To compare the isolated colchicine with pure standard colchicine, we used the H-NMR technique. The methyl thiazolyl tetrazolium (MTT) assay, quantitative reverse transcriptase-polymerase chain reaction, Western blotting and annexin V/PI staining were used to evaluate the apoptotic effects of the isolated and standard colchicine. RESULTS: Similar to standard colchicine, the isolated colchicine inhibited cell proliferation significantly in cancer cell lines. Colchine inhibited proliferation and induced apoptosis on a dose-dependent manner. The medicine modified the expression of genes-related to apoptosis by up-regulation of P53 ,BAX, CASPASE-3, -9 and down-regulation of BCL-2 gene, which led to an increase in the BAX/BCL-2 ratio. CONCLUSION: We showed that isolated colchicine from Colchicum autumnale and pure standard colchicines modulate the expression levels of several genes and therefore exerting their anticancer effects on both human (MCF-7) and mouse (4T1) breast cancer cells. Based on these results, we suggest that colchicine can be a potential candidate for prevention and treatment of breast cancer.

Differentiating Cannabis Products: Drugs, Food, and Supplements.

"Hemp" refers to non-intoxicating, low delta-9 tetrahydrocannabinol (Δ9-THC) cultivars of Cannabis sativa L. "Marijuana" refers to cultivars with high levels of Δ9-THC, the primary psychoactive cannabinoid found in the plant and a federally controlled substance used for both recreational and therapeutic purposes. Although marijuana and hemp belong to the same genus and species, they differ in terms of chemical and genetic composition, production practices, product uses, and regulatory status. Hemp seed and hemp seed oil have been shown to have valuable nutritional capacity. Cannabidiol (CBD), a non-intoxicating phytocannabinoid with a wide therapeutic index and acceptable side effect profile, has demonstrated high medicinal potential in some conditions. Several countries and states have facilitated the use of THC-dominant medical cannabis for certain conditions, while other countries continue to ban all forms of cannabis regardless of cannabinoid profile or low psychoactive potential. Today, differentiating between hemp and marijuana in the laboratory is no longer a difficult process. Certain thin layer chromatography (TLC) methods can rapidly screen for cannabinoids, and several gas and liquid chromatography techniques have been developed for precise quantification of phytocannabinoids in plant extracts and biological samples. Geographic regulations and testing guidelines for cannabis continue to evolve. As they are improved and clarified, we can better employ the appropriate applications of this uniquely versatile plant from an informed scientific perspective.

Glucosinolates and their hydrolysis products as potential nutraceuticals to combat cytokine storm in SARS-COV-2.

INTRODUCTION: The high mortality rate in severe cases of COVID-19 is mainly due to the strong upregulation of cytokines, called a cytokine storm. Hyperinflammation and multiple organ failure comprise the main clinical features of a cytokine storm. Nrf2 is a transcription factor which regulates the expression of genes involved in immune and inflammatory processes. Furthermore, Nrf2, as a master regulator, controls the activity of NF-κB which binds to the promoter of many pro-inflammatory genes inducible of various inflammatory factors. Inhibition of Nrf2 response was recently demonstrated in biopsies from patients with COVID-19, and Nrf2 agonists inhibited SARS-CoV-2 replication across cell lines in vitro. Glucosinolates and their hydrolysis products have excellent anti-inflammatory and antioxidant effects via the Nrf2 activation pathway, reduction in the NF-κB activation, and subsequent reduced cytokines levels. CONCLUSION: Accordingly, these compounds can be helpful in combating the cytokine storm associated with COVID-19.

Salvigenin, a Trimethoxylated Flavone from Achillea Wilhelmsii C. Koch, Exerts Combined Lipid-Lowering and Mitochondrial Stimulatory Effects.

Phytochemical analysis of the Iranian plant Achillea wilhelmsii led to the isolation of 17 pure secondary metabolites belonging to the classes of sesquiterpenoids and phenolics. Two of these compounds, named wilhemsin (7) and wilhelmsolide (9), are new sesquiterpenoids, and the first shows undescribed structural features. Their structures were elucidated through extensive spectroscopic analysis, mainly based on 1D and 2D NMR, and chemical derivatization. Starting from plant traditional use and previous reports on the activity of the plant extracts, all the pure compounds were evaluated on endpoints related to the treatment of metabolic syndrome. The sesquiterpene hanphyllin (8) showed a selective cholesterol-lowering activity (-12.7% at 30 µM), santoflavone (13) stimulated glucose uptake via the GLUT transporter (+16.2% at 30 µM), while the trimethoxylated flavone salvigenin (14) showed a dual activity in decreasing lipid levels (-22.5% palmitic acid biosynthesis at 30 µM) and stimulating mitochondrial functionality (+15.4% at 30 µM). This study further confirms that, in addition to the antioxidants vitexin, isovitexin, and isoschaftoside, A. wilhelmsii extracts contain molecules that can act at different levels on the metabolic syndrome symptoms.

Comparison of the cytotoxic effects of different fractions of Artemisia ciniformis and Artemisia biennis on B16/F10, PC3 and MCF7 Cells.

BACKGROUND AND PURPOSE: Artemisia is one of the well-known herbal medicinal plants for antimicrobial, insecticidal, antioxidant, and antimalarial activities. The antiproliferative effects of dichloromethane extracts of Artemisia biennis (A. biennis) and A. ciniformis and the petroleum ether extract of A. ciniformis have been demonstrated previously on human cancerous cell lines. In the current study, further fractionation was carried out on the aforementioned extracts and their cytotoxic effects were evaluated on three human cancer cell lines; B16/F10, PC3, and MCF7. F1 to F16, F1' to F11', and F1" to F10" were resulted from the fractionation of dichloromethane extracts of A. biennis, A. ciniformis, and petroleum ether extract of A. ciniformis, respectively. EXPERIMENTAL APPROACH: The cytotoxic effects of 16 (F1-F16), 11 (F1'-F11') and 10 (F1"-F10") fractions, on B16/F10, PC3, and MCF7 cell lines were assessed using resazurin to measure viability and propidium iodide staining (sub G1) and flow cytometry to detect apoptosis. FINDINGS / RESULTS: The results showed that, some fractions at 100 µg/mL decreased cell viability. F2" in B16/F10 cells, F2, F4-F6, F10', F11', and F2" in PC3 cells, and F10', F11', and F2" in MCF7 significantly decreased cell viability in a concentration-dependent manner (12.5-50 µg/mL). Among different fractions, F2" demonstrated the most potent cytotoxic effects on cancer cell lines (P < 0.001). All of the mentioned fractions (except F11' on PC3 cells) increased the number of apoptotic cells and showed the cytotoxic effects on cancer cells compared with the control group. CONCLUSION AND IMPLICATIONS: A. biennis and A. ciniformis are suggested as the potential sources of cytotoxic phytochemicals. The probable presence of terpenoids, steroids, and alkaloids in the selected fractions is proposed based on the preliminary phytochemical study.

Antidiabetic Potential of Prosopis farcta Roots: In Vitro Pancreatic Beta Cell Protection, Enhancement of Glucose Consumption, and Bioassay-Guided Fractionation.

By using the streptozotocin- (STZ-) induced cytotoxicity in ß-TC3 cells as an assay model, a bioassay-guided fractionation study was employed to isolate and characterize the potential antidiabetic principles of roots of Prosopis farcta. A combination of open column chromatography on reverse-phase silica gel using a water-ethanol gradient (10 : 90 to 100 : 0) followed by HPLC-based fractionation led to an active compound that appears to be composed of carbohydrate/sugar. When cell viability under STZ was reduced to 49.8 ± 4% (mean ± SD), treatment with the active compound at the concentration of 0.5 mg/mL either as a coadministration or a pretreatment improved the viability to 93 ± 1.9% and 91.5 ± 7%, respectively. The reduction in the mitochondrial membrane potential by STZ (47.34 ± 8.9% of control) was similarly recovered to 84.5 ± 4.3 (coadministration) and 88 ± 5.5% (pretreatment) by the active fraction. The bioassay-guided fractionation, ß-cell protective effect, and increased glucose consumption (up to 1.49-fold increase) in hepatocytes by the extracts and active fraction are also discussed.

Efficacy and safety of Achillea wilhelmsii C. Koch capsules on symptom severity and quality of life in patients with irritable bowel syndrome: a randomized, placebo-controlled clinical trial.

Background Irritable bowel syndrome (IBS) is one of the most common digestive diseases. The aim of this clinical trial was to determine the effectiveness of Achillea wilhelmsii C. Koch on the symptom severity and quality of life (QOL) in patients with IBS. Methods The patients were randomized into two groups of 45 each. The QOL and symptom severity of the patients were evaluated at baseline and at completion of the treatments by means of IBS-QOL and IBS severity index. Results The mean severity of clinical symptoms in the Achillea wilhelmsii C. Koch receiving groups before and after the treatment was 282.56 ± 103.57 and 178.06 ± 88.40, and in the placebo group was 265.93 ± 93.56 and 197.74 ± 106.26, respectively. The mean QOL in the Achillea wilhelmsii C. Koch receiving group before and after treatment was 51.49 ± 11.98 and 50.44 ± 13.39 and in the placebo group was 60.71 ± 11.97 and 58.39 ± 11.67, respectively. In both groups, there was a significant difference in the recovery rate in each group (p<0.05). However, the mean difference between the two groups before and after intervention was not significantly different (p>0.05). Also, no patient reported any adverse events during the trial. Although the symptom severity and QOL in both groups were improved compared to those before intervention, there was no significant difference between the two groups. Conclusion It is recommended to conduct future studies with larger sample size and longer treatment periods, and also investigate the efficacy on the IBS subtypes, separately.

Efficacy and safety of a standardized extract from Achillea wilhelmsii C. Koch in patients with ulcerative colitis: A randomized double blind placebo-controlled clinical trial.

BACKGROUND: Using Achillea wilhelmsii as a dietary supplement for gastrointestinal disorders is common in Persian traditional medicine. Its anti-inflammatory, anti-spasmodic and antibacterial properties have been proven by different in vitro and in vivo studies, yet it has not been evaluated in a controlled clinical trial. AIM: This study intended to evaluate the efficacy and safety of A. wilhelmsii in patients with mild to moderate active ulcerative colitis in a randomized, double-blinded, placebo-controlled clinical trial. The hydroalcoholic extract of A. wilhelmsii was standardized based on caffeic acid. METHODS: Forty-nine patients were randomly received A. wilhelmsii capsules or placebo, twice daily for 4 weeks in a 1:1 ratio. The disease activity index (DAI) (Partial Mayo Score), haemoglobin, platelet count, erythrocyte sedimentation rate (ESR) and serum level of C-reactive protein (CRP) were measured at the entry and the end of the treatment. To standardize the extract, caffeic acid was detected and measured in the plant extract using high performance liquid chromatography (HPLC). RESULTS: Of 49 patients who entered the trial, 40 patients completed the study. In both treatment and placebo groups, significant reductions were observed in stool frequency, rectal bleeding, physician global assessment and partial mayo score. There was no significant difference in stool frequency (P = 0.176), rectal bleeding (P = 0.523), physician global assessment (P = 0.341) and partial mayo score (P = 1) in the treatment versus the placebo groups. Laboratory variables including hemoglobin, platelet count, ESR and CRP showed no significant difference between the treatment and the placebo group. Of all participants, only one patient in the treatment group complained about skin rash (grade 1 based on the Common Terminology Criteria for Adverse Events (CTCAE) version 4.0). CONCLUSION: Oral administration of A. wilhelmsii powder for 4 weeks did not create a clinical response more than placebo. It seemed to be safe in UC patients. Further studies are obligatory to evaluate the therapeutic potential of A. wilhelmsii in the form of extract in UC patients.

Mechanistic In vitro Evaluation of Prosopis farcta Roots Potential as an Antidiabetic Folk Medicinal Plant.

OBJECTIVE: Prosopis farcta has been used as a traditional herbal medicine for treating Diabetes mellitus. The aim of this study is to investigate the antidiabetic mechanisms of infusion (INF) extract of P. farcta and discovering the active extract for the first time. MATERIALS AND METHODS: Six different extracts of P. farcta were prepared using five different solvents (ethanol, n-hexane, acetone, ethanol:water (1:1 v/v), and water). Cytotoxicity and cell proliferation assays were performed on mouse pancreatic ß-cells (ß-TC3) using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium method. The effects of P. farcta on glucose metabolism (in a hepatocellular carcinoma cell line [HepG2]) and glucose diffusion across a dialysis membrane (as a model of cellular glucose absorption) were evaluated. The protective effect of various P. farcta extracts on cytotoxicity, mitochondrial membrane potential (MMP), and streptozotocin (STZ)-induced apoptosis in ß-TC3 cells was investigated. RESULTS: Cytotoxicity study indicated that extracts were safe on ß-TC3 and HepG2 (≤0.5 mg/ml). INF protected ß-TC3 cells from apoptosis induced by STZ and improved cell viability for 20% and significantly decrease depolarization of MMP (P < 0.005). The results showed that INF inhabited breaking/streaking the DNA. Proliferation study showed no significant increase in the number of cells either at single or multiple doses. In moderate hyperglycemia (11.1 mmol/l), a significant glucose-lowering effect was observed but glucose diffusion was not the probable mechanism of extracts antidiabetic effect. In conclusion, only INF, the traditionally used extract, has an antidiabetic potential by attenuating the death and apoptosis induced by STZ in ß-TC3 cells and increase glucose consumption. CONCLUSION: The present study demonstrates that only INF extract have an antidiabetic potential by attenuating the death and apoptosis induced by STZ in ß-TC3 cells and increase glucose consumption. SUMMARY: Six different extracts from P. farcta were prepared using five different solvents [ethanol, n-hexane, acetone, ethanol: water (1:1 v/v), and water]The protective effect of various P. farcta extracts on cytotoxicity, mitochondrial membrane potential (MMP), and Streptozotocin-induced apoptosis in ß-TC3 cells were investigated.Infusion has an antidiabetic potential by attenuating the death and apoptosis induced by STZ in ß-TC3 cells and increase glucose consumptionThe effect of infusion extract on glucose consumption in hepatocellular carcinoma cell line cells (a) and effect of infusion extract on glucose consumption in hepatocellular carcinoma cell line cells adjusted by optical density MTT (b). Significance was calculated by analysis of variance (*P ≤ 0.05). MTT: 3 (4,5 dimethylthiazol 2 yl) 2,5 diphenyltetrazolium. Abbreviations used: AC: Acetone extract; ANOVA: Analysis of variance; BSA: Bovine serum albumin; ß-TC3: Mouse pancreatic ß-cells; DMEM: Dulbecco modified Eagle medium; DMSO: Dimethyl sulfoxide; ETH: Ethyl acetate extract; FBS: Fetal bovine serum; HDETH: Hydroethanolic extract; HepG2: Hepatocellular carcinoma cell line; HEX: Hexane extract; INF: Infusion; KUMS: Kermanshah University of Medical Sciences; MMP: Mitochondrial membrane potential; MTT: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium; NaCl: Natrium chloride; OD: Optical density; spp: Species; STZ: Streptozotocin; Tag: T-antigen; USA: United States of America.

Bioassay-guided Isolation of Neuroprotective Fatty Acids from Nigella sativa against 1-methyl-4-phenylpyridinium-induced Neurotoxicity.

OBJECTIVE: Parkinson's disease, a slowly progressive neurological disease, is associated with degeneration of the basal ganglia of the brain and a deficiency of the neurotransmitter dopamine. The main aspects of researches are the protection of normal neurons against degeneration. Fatty acids (FAs), the key structural elements of dietary lipids, are carboxylic straight chains and notable parameters in nutritional and industrial usefulness of a plant. MATERIALS AND METHODS: Black cumin, a popular anti-inflammatory and antioxidant food seasoning, contains nonpolar constituents such as FAs which were extracted using hexane. Different fractions and subfractions were apt to cytoprotection against apoptosis and inflammation induced by 1-methyl-4-phenylpyridinium (MPP+) in rat pheochromocytoma cell line (PC12) as a neural cell death model. The experiment consisted of examination of cell viability assessment, mitochondrial membrane potential (MMP), caspase-3 and -9 activity, and measurement of cyclooxygenase (COX) activity. RESULTS: MPP+ induced neurotoxicity in PC12 cells. Pretreatment with subfractions containing FA mixtures attenuated MPP+-mediated apoptosis partially dependent on the inhibition of caspase-3 and -9 activity and increasing the MMP. A mixture of linoleic acid, oleic acid, and palmitic acid also decreased the COX activity induced by MPP+ in PC12 cells. CONCLUSION: Our observation indicated that subtoxic concentration of FA from Nigella sativa may exert cytoprotective effects through their anti-apoptotic and anti-inflammation actions and could be regarded as a dietary supplement. SUMMARY: MPP+ induced neurotoxicity in PC12 cellsNigella sativa contains bioactive fatty acidsPretreatment with fatty acids attenuated MPP+ mediated apoptosis through inhibition of caspase 3 and 9 activityA mixture of linoleic acid, oleic acid, and palmitic acid decreased the COX activity induced by MPP+ in PC12 cellsDue to cytoprotective, anti apoptotic and anti inflammation actions of N. sativa, it could be regarded as a dietary supplement. Abbreviations used: ANOVA: Analysis of variance; Ca: Calcium; CDCl3: Chloroform; COX: Cyclooxygenase; DMSO: Dimethyl sulfoxide; EA: Elidic acid; EDTA: Ethylene diamine tetraacetic acid; ELISA: Enzyme Linked Immunosorbent Assay; ESI-MS: Electron spray mass spectroscopy; FAs: Fatty acids; FBS: Fetal bovine serum; GC: Gas chromatography; 1HNMR: Hydrogen nuclear magnetic resonance; LA: Linoleic acid; MPP+: 1-Methyl-4-phenylpyridinium; MPTP: 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine; MTT: 3-(4,5-dimethylthiazol-2yl)-2,5-diphenyltetrazolium bromide; N. sativa: Nigella sativa; OA: Oleic acid; PA: Palmitic acid; PBS: Phosphate buffer saline; PC12: Rat pheochromocytoma cell line; PD: Parkinson's disease; PDA: Photo diode array detector; PGE2: Prostaglandin E2; TLC: Thin layer chromatography; TMPD: N,N,N',N'-tetramethyl-p-phenylenediamine; USA: United states of America.

Ethanolic Extract of Berberis Vulgaris Fruits Inhibits the Proliferation of MCF-7 Breast Cancer Cell Line Through Induction of Apoptosis

BACKGROUND: As it is obvious, there is much documentation that shows the importance of breast cancer treatment in patients. High expressions of P53 and Bcl-2 are associated with breast cancer, which are reliable factors to follow up the breast cancer. Berberis vulgaris is used as a traditional medicine in cancer. Despite of the fact that many researches have demonstrated its anti-cancer properties, there are no scientific documents to show its efficacy in detail in breast cancer. OBJECTIVE: Because of traditional use of B. vulgaris and little knowledge about its effects, our research was focused on determining the efficacy and toxicity of B. vulgaris. For this reason, we determined the efficacy of B. vulgaris on breast cancer cells. METHOD: As described in Method section, standard protocols including MTT assay and qPCR were performed to identify the effect of B. vulgaris ethanolic extract against breast cancer cells. RESULTS: Our results clearly demonstrated that 35 mg/ml had IC50 against 3t3 normal cells, and 9 mg/ml of B. vulgaris was effective against MCF-7 breast cancer cells. The results demonstrated that even at only 1 mg/ml concentration of B. vulgaris, crude extract was effective, 9 mg/ml and 12 mg/ml of extract had better anti-cancer activity compared with doxorubicin. CONCLUSION: Despite that the role of anticancer properties of B. vulgaris was clearly defined in some patents, our results demonstrated the potency of B. vulgaris against breast cancer, but further analysis should be performed to candidate this herb as an anti-breast cancer drug.

New polyacetylenes from Echinophora cinerea (Boiss.) Hedge et Lamond.

Echinophora cinerea aerial parts are used in folk medicine to cure gastric diseases and as a food seasoning in cheese and yogurt. Besides several pharmacological effects have been assigned to Echinophora spp., there is no phytochemical investigation on this genus other than our previous publication on flavonoids. An acetone extract of E. cinerea afforded three new (1-3) polyacetylenes, one rare monoterpenoid glycoside as verbenone-5-O-ß-D-glycopyranoside (4) and one prenylated coumarin as osthol (5). The structures of all new compounds were elucidated using modern spectroscopic methods, including 2D NMR and mass analyses. The potency of the compounds to induce cell death was determined on SKNMC, PC3 and MCF-7 cell lines using MTT method in which compounds 1 and 2 showed moderate cytotoxic effects, especially against PC3 cells.

Anti-Viral Evaluation of Sesquiterpene Coumarins from Ferula assa-foetida against HSV-1.

Several complications attributed with Herpes virus related infections and the emergence of drug resistant viruses prompt scientists to search for new drugs. Several terpenoids and coumarins have shown anti HSV effects while no sesquiterpene coumarins have been previously tested for HSV treatment. Three sesquiterpene coumarins badrakemin acetate (1), kellerin (2) and samarcandin diastereomer (3) were isolated from the gum resin of Ferula assa-foetida, a herbal medicine with antimicrobial, antiprotozoal and antiviral effects. Compounds were identified by 1D and 2D- NMR spectroscopies and comparison with literature data. A comparative evaluation of cytotoxicity and antiviral activity showed that kellerin (2) could significantly inhibit the cytopathic effects and reduce the viral titre of the herpes virus type 1 (HSV-1) DNA viral strain KOS at concentrations of 10, 5 and 2.5 µg/mL.

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.

Antiviral and cytotoxic evaluation of coumarins from Prangos ferulacea.

CONTEXT: Prangos ferulacea (L.) Lindl. (Apiaceae) is a perennial plant found in the Middle-East, where it is commonly used as an antispasmodic and anti-inflammatory agent. It is a rich source of coumarins. OBJECTIVE: To purify several coumarins from P. ferulacea and to screen their cytotoxicity and anti-herpes activity. MATERIALS AND METHODS: Acetone extract of roots of P. ferulacea was subjected to several chromatographic separations to render pure coumarins (1-8). Anti-herpes virus effects of 1-7 were evaluated at concentration 2.5, 5, and 10 µgmL(-1), on a confluent monolayer of Vero cells infected with 25 PFU of HSV1. Cytotoxic effects of 1 and 2 were evaluated on an A2780S cell line using the MTT assay. The cells were exposed to a series of concentrations of coumarins (0.01-2.5 mM, 37°C, 72 h). RESULTS: Compounds 1-8 were identified as osthole, isoimperatorin, oxypeucedanin, psoralen, oxypeucedanin hydrate, gosferol, oxypeucedanin methnolate, and pranferol. This is the first report of occurrence of 4 and 7 in this plant. Compound 1 showed a viability of 9.41% ± 2.4 at 2.5 mM on A2780S cells (IC50 = 0.38 mM). The cell survival of 2 at 2.5 mM was 46.86% ± 5.5 with IC50 equal to 1.1 mM. DISCUSSION AND CONCLUSION: Compound 1 shows cytotoxic effects on the A2780S cell line. Compound 2 is a cyclooxygenase-2 inhibitor and the A2780S cell line does not express COX-2 which may interpret the non-toxic effect of the compound on this cell line. None of the tested compounds showed an anti-HSV effect at non-toxic concentrations.

Antiproliferative evaluation of terpenoids and terpenoid coumarins from Ferulago macrocarpa (Fenzl) Boiss. fruits.

BACKGROUND: Ferulago macrocarpa is a plant used as flavoring agent and protectant in the food industry and as a folk medicinal plant in Iran with no available information on its chemical identity. Ferulago spp. showed to contain biologically terpenoids and coumarins. OBJECTIVE: The objective was to isolate and characterize terpenoids and coumarins from the acetone extract of F. macrocarpa fruits and to evaluate their antiproliferative effects on several cell lines. MATERIALS AND METHODS: A series of normal and reverse phase gravity and high-performance liquid chromatography analyses were used to purify constituents. Compounds 1-5 and 7 were evaluated for their cytotoxic effects on MCF-7, HT-29 and H-1299 cell lines. RESULTS: Six compounds including bornyl acetate (1), 1,10-di-epi-cubenol (2), stigmasterol (3) and three coumarins grandivittin (4), prantschimgin (5) and 4"-hydroxygrandivittin (7) along with mixtures of feruloyl derivatives (6a-6c) have been purified. Their structures were established by spectroscopic methods including nuclear magnetic resonance and MS analyses. Compound 2 showed moderate cytotoxicity effect with IC50 values of 5.0 and 6.7 mM on MCF-7 and HT-29, respectively. CONCLUSION: 1,10-di-epi-Cubenol could be considered as a potential proliferation inhibitor of MCF-7 and HT-29 cell lines.

Some like it pungent and vile. TRPA1 as a molecular target for the malodorous vinyl disulfides from asafoetida.

Ferula assa-foetida L. is a major source of asafoetida, a foul-smelling gum-resin of dietary and medicinal relevance. Investigation of the roots and latex of F. assa-foetida yielded various mixed vinyl disulfides, including two novel asadisulfide-type esters. The reactivity of asadisulfide was investigated in the cysteamine NMR assay, where it behaved as disulfide shuffling agents due to the high mobility of the vinyl sulfide moiety. In accordance with their transthiolation capacity, these compounds potently activated TRPA1, the target of mustard oil and the pungent sulfur compounds from onion and garlic, qualifying S-alkyl-S-alkenyldisulfides, a rare class of natural products, as a novel class of dietary TRPA1 activators. These observations provide a mechanistic basis for rationalizing some of the sensory properties and potential beneficial health claims associated to the use of asafoetida as a spice and as a medicine.

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.

Isolation and Identification of Ferulic Acid From Aerial Parts of Kelussia odoratissima Mozaff.

BACKGROUND: Kelussia odoratissima Mozaff. is one of the newest genera of Umbelliferae which is represented by only one species. This sweet-smelling, self-growing monotypic medicinal plant is endemic to a restricted area in west of Iran and is locally called Karafse-koohi. The aerial parts of the plant are commonly used as a popular garnish and a sedative medicinal plant. There are several reports concerning antioxidant, anti-inflammatory, anxiolytic and hypolipidemic activities of aerial parts of K. odoratissima. OBJECTIVES: The current research aimed to evaluate some phenolic contents of the plant for the first time .It is confirmed that secondary metabolites and especially phenolic compounds have important role in the biological activities of the plant. Available information indicates that phenolic contents of K. odoratissima have not been the subject of any investigation. MATERIAL AND METHODS: Aerial parts of K. odoratissima were extracted with acetone by maceration method. Normal and reversed phase vacuum liquid chromatography used to fractionate the extract. (1)H-NMR, (13)CNMR, EI-Mass and IR spectra were used to elucidate isolated compound. RESULTS: The phenolic acid isolated compound was identified as 4-hydroxy-3-methoxycinnamic acid (ferulic acid). CONCLUSIONS: Compared with previous reported antioxidant and anti-inflammatory properties of ferulic acid, a chemical-biological relation can be postulated.