OBJECTIVETo study the effect of Flos Daturae alkaloids (FDA) on TGF-beta1-1uuuu;U epithelial-mesenchymal transition (EMT) of human pulmonary adenocarcinoma A549 cells.

METHODSA549 cells in vitro cultured were randomly divided into 5 groups, i.e., the blank control group, the TGF-beta1 group, the low dose FDA group, the medium dose FDA group, and the high dose FDA group. The morphologies of A549 cells were observed. Expressions of cytokeratin (CK)-19 and alpha-smooth muscle actin (alpha-SMA) were detected by Western blot and real-time PCR at 24, 48, and 72 h, respectively.

RESULTSA549 cells in the TGF-beta1, group turned from cobblestone to spindle shape gradually. Those in low, medium and high dose FDA groups showed similar shapes to those of the TGF-beta1 group. There was no statistical difference in the morphology of A549 cells among the 3 dose FDA groups (P > 0.05). Western blot showed that, when compared with the blank control group, the expression of CK-19 was down-regulated, but the expression of alpha-SMA was up-regulated in the TGF-beta1 group (P < 0.01). Compared with the TGF-beta1, group, the expression of CK-19 was up-regulated, but the expression of alpha-SMA was suppressed in low, medium and high dose FDA groups (P < 0.01). The CK-19 expression obviously increased, but the alpha-SMA expression was suppressed in high dose FDA group at 72 h (P < 0.01). Real-time PCR results showed, as compared with the TGF-beta1 group, the mRNA expression of CK-19 was increased, but the mRNA expression of alpha-SMA was reduced in low, medium and high dose FDA groups (P < 0.01).

CONCLUSIONSFDA had no effect on EMT morphological changes of TGF-beta1 induced A549 cells. FDA could reverse characteristic markers of A549 cells during EMT to some extent, such as expressions of CK-19 and alpha-SMA. The expression of CK-19 (as the epithelium marker) increased and the expression of alpha-SMA (as the mesenchymal marker) was reduced. Besides, they were most obviously seen in the high dose FDA group at 72 h in a dose- and time-dependent manner.

Actins ; Adenocarcinoma ; Alkaloids ; pharmacology ; Cell Line, Tumor ; Datura ; Drugs, Chinese Herbal ; pharmacology ; Epithelial Cells ; Epithelial-Mesenchymal Transition ; Epithelium ; Humans ; Transforming Growth Factor beta1 ; metabolism

Hyoscyamine 6 beta-hydroxylase (H6H) is the last rate-limiting enzyme directly catalyzing the formation of scopolamine in tropane alkaloids (TAs) biosynthesis pathway. It is the primary target gene in the genetic modification of TAs metabolic pathway. Full-length cDNA and gDNA sequences of a novel H6H gene were cloned from Datura arborea (DaH6H, GenBank accession numbers for cDNA and gDNA are KR006981 and KR006983, respectively). Nucleotide sequence analysis reveals an open reading frame of 1375 bp encoding 347 amino acids in the cDNA of DaH6H, while the gDNA of DaH6H contains four exons and three introns, with the highest similarity to the gDNA of H6H from D. stramonium. DaH6H also exhibited the most identity of 90.5% with DsH6H in amino acids and harbored conserved 2-oxoglutarate binding motif and two iron binding motifs. The expression level of DaH6H was highest in the mature leaf, followed by the secondary root, and with no expression in the primary root based on qPCR analysis. Its expression was inhibited by MeJA. DaH6H was expressed in E. coli and a 39 kD recombinant protein was detected in SDS-PAGE. Comparison of the contents of scopolamine and hyoscyamine in various TAs-producing plants revealed that D. arborea was one of the rare scopolamine predominant plants. Cloning of DaH6H gene will allow more research in the molecular regulatory mechanism of TAs biosynthesis in distinct plants and provide a new candidate gene for scopolamine metabolic engineering.
Cloning, Molecular ; DNA, Complementary ; Datura ; enzymology ; genetics ; Escherichia coli ; Hyoscyamine ; chemistry ; Mixed Function Oxygenases ; genetics ; Plant Leaves ; enzymology ; Plant Roots ; enzymology ; Recombinant Proteins ; genetics ; Scopolamine Hydrobromide ; chemistry

AIM: The anti-leishmanial activity of methanolic extracts of Calendula officinalis flowers, Datura stramonium seeds, and Salvia officinalis leaves against extracellular (promastigote) and intracellular (amastigote) forms of Leishmania major were evaluated in this study. METHOD: In the first stage, promastigote forms of L. major, were treated with different doses of the plant extracts in a 96-well tissue-culture microplate and IC50 values for each extract were measured with colorimetric MTT assay. In the second stage, macrophage cells were infected with L. major promastigotes. Infected macrophages were treated with plant extracts. Then the macrophages were stained with Gimsa and the number of infected macrophages and amastigotes were counted with a light microscope. RESULTS: The results indicated that the plant extracts inhibited the growth of promastigotes and amastigotes of L. major. Inhibitory concentrations (IC50) for promastigote assay were 108.19, 155.15, and 184.32 μgmL(-1) for C. officinalis flowers, D. stramonium seeds and S. officinalis, respectively. The extracts also reduced the number of amastigotes in macrophage cells from 264 for control group to 88, 97, and 102 for test groups. Although the anti-leishmanial activity of the extracts were not comparable with the standard drug, miltefosine; but they showed significant efficiency in reducing the number of amastigotes in macrophages, in comparison with the control group (P < 0.001). These plant extracts had lower toxicity compared with miltefosine. CONCLUSION This study demonstrates the potential efficacy of the methanolic extracts of C. officinalis flowers, D. stramonium seeds, and S. officinalis leaves to control of cutaneous leishmaniasis.
Antiparasitic Agents ; pharmacology ; therapeutic use ; Calendula ; Cell Line ; Datura stramonium ; Flowers ; In Vitro Techniques ; Leishmania major ; drug effects ; Leishmaniasis ; drug therapy ; parasitology ; Macrophages ; drug effects ; parasitology ; Phytotherapy ; Plant Extracts ; pharmacology ; therapeutic use ; Plant Leaves ; Salvia officinalis ; Seeds

OBJECTIVETo study the effects of methyl jasmonate (MJ) on the accumulation and release of main secondary metabolites i. e. scopolamine and hyoscyamine in liquid cultures of Datura stramonium hairy roots.

METHODAfter 18 days liquid culture of D. stramonium hairy roots induced by agrobacterium rhizogenes C58C1, the chemical elicitor methyl jasmonate was added into 1/2 MS liquid cultures and scopolamine and hyoscyamine on the day 0, 3, 6, 9 and 12, after dealing with MJ, was determined by HPLC.

RESULTAfter dealing with MJ on the day 3, 6, 9 and 12,the concentration of scopolamine reached to 0.419, 0.439, 0.431, 0.374 mg x g(-1), respectively, the increase of scopolamine were 1.36, 1.42, 1.17 and 1.12 fold higher than that of the control, respectively. And hyoscyamine reached 1.493, 0.817, 0.723 and 0.698 mg x g(-1), respectively, the increase of hyoscyamine were 2.28, 1.11, 0.63 and 0.70 fold higher than that of the control, respectively.

CONCLUSIONMJ could stimulate the accumulation of scopolamine and hyoscyamine (3,6 d) in D. stramonium hairy root and have released them into the culture medium.

Acetates ; pharmacology ; Alkaloids ; analysis ; metabolism ; Cell Culture Techniques ; Cyclopentanes ; pharmacology ; Datura stramonium ; chemistry ; drug effects ; growth & development ; metabolism ; Oxylipins ; pharmacology ; Plant Roots ; chemistry ; drug effects ; growth & development ; metabolism ; Tropanes ; analysis ; metabolism

Datura stramonium L., a wild-growing plant of the Solanaceae family, is widely distributed and easily accessible. It contains a variety of toxic tropane alkaloids such as atropine, hyoscamine, and scopolamine. In Eastern medicine, especially in Ayurvedic medicine, D. stramonium has been used for curing various human ailments, including ulcers, wounds, inflammation, rheumatism and gout, sciatica, bruises and swellings, fever, asthma and bronchitis, and toothache. A few previous studies have reported on the pharmacological effects of D. stramonium; however, complete information regarding the pharmacology, toxicity, ethnobotany and phytochemistry remains unclear. Ethnomedicinally, the frequent recreational abuse of D. stramonium has resulted in toxic syndromes. D. stramonium, in the form of paste or solution to relieve the local pain, may not have a deleterious effect; however, oral and systemic administration may lead to severe anticholinergic symptoms. For this reason, it is very important for individuals, mainly young people, to be aware of the toxic nature and potential risks associated with the use of this plant. This comprehensive review of D. stramonium includes information on botany, phytochemistry, pharmacology, toxicology and ethnomedicinal uses.
Animals ; Datura ; chemistry ; classification ; Humans ; Phytotherapy ; Plant Extracts ; chemistry ; pharmacology ; toxicity

OBJECTIVETo establish the hair roots culture system of Datura metel and study the hair roots growth and biosynthesis of scopolamine and hyoscyamine in hair roots culturing system.

METHODDirect degermed cotyledon of wild D. metel was infected by Agrobacterium tumefaciens strain C58C1 to obtain hair roots. Growth curves and scopolamine and hyoscyamine biosynthesis curves were determined. The scopolamine and hyoscyamine from different hair roots lines were examined by HPLC.

RESULTHair roots induction rate reached 70%. After 25 days cultured in 1/2 MS liquid nutrient medium, the hair roots weight, content of scopolamine and hyoscyamine reached maximum, tow high efficient accumulation hyoscyamine and scopolamine hair roots lines M1 and M2 were obtained. The medial accumulation coefficient of hyoscyamine and scopolamine were 2.53 times and 5.37 times compared with the leaves of wild D. metel respectively.

CONCLUSIONThe established hair roots induction and culture system of D. metel provided a foundation for further obtaining scopolamine and hyoscyamine.

Atropine ; analysis ; biosynthesis ; Datura metel ; chemistry ; growth & development ; metabolism ; Plant Roots ; chemistry ; growth & development ; metabolism ; Scopolamine Hydrobromide ; analysis ; metabolism

India has a great wealth of various naturally occurring plant drugs which have great potential pharmacological activities. Datura stramonium (D. stramonium) is one of the widely well known folklore medicinal herbs. The troublesome weed, D. stramonium is a plant with both poisonous and medicinal properties and has been proven to have great pharmacological potential with a great utility and usage in folklore medicine. D. stromonium has been scientifically proven to contain alkaloids, tannins, carbohydrates and proteins. This plant has contributed various pharmacological actions in the scientific field of Indian systems of medicines like analgesic and antiasthmatic activities. The present paper presents an exclusive review work on the ethnomedical, phytochemical, pharmacological activities of this plant.
Anti-Asthmatic Agents ; Anti-Bacterial Agents ; Datura stramonium ; chemistry ; toxicity ; Ethnopharmacology ; Humans ; India ; Medicine, Traditional ; Phytotherapy ; Plant Extracts ; chemistry ; pharmacology ; Plants, Medicinal ; chemistry ; toxicity ; Trees ; chemistry ; toxicity

OBJECTIVETo study chemical constituents in the seeds of Datura Stramonium (Solanaceae family).

METHODCompounds were isolated and purified by silica gel, MCI and Sephadex LH-20 column chromatography, and their structures were determined based on physicochemical constants and spectroscopic analysis including NMR and MS.

RESULTTwelve compounds were isolated and identified from Datura stramonium, they were N-trans-feruloyl tryptamine (1), hyoscyamilactol (2), scopoletin (3), umckalin (4), daturaolone (5), daturadiol (6), N-trans-ferulicacyl- tyramine (7), cleomiscosin A (8), fraxetin (9), scopolamine (10), 1-Acetyl-7-hydrox-beta-carbol-ine (11), 7-hydroxy-beta-carbolinel-propionic acid (12).

CONCLUSIONCompound 2, 7, 9 and 12 were obtained from Datura genus for the first time, whereas compound 1, 4, 8 and 11 were obtained from the Solanaceae family for the first time.

Datura stramonium ; chemistry ; Seeds ; chemistry

To identify the original plant of Daturae Flos from its adulterants by DNA barcoding, the sequences of ITS2, psbA-trnH, matK, rbcL of four species including Datura metel, Darura innoxia, Darura stramonium and Brugmansia arborea were compared and analyzed. The PCR and sequencing success rate of the four regions (ITS2, psbA-trnH, matK, rbcL) was 100%, 90%, 100% and 85%, respectively. Sequences were assembled with CodonCode Aligner. K2P distances were calculated and NJ tree was performed by MEGA 4.1. Thirty SNPs were found among ITS2 sequences, and 33 insert/deletes were found among psbA-trnH intergenic regions. The interspecific K2P distance of ITS2 and psbA-trnH was obviously higher than that of the intraspecific one. As to matK and rbcL, there was no "Barcoding Gap" existing between inter- and intra-specific distances. The NJ trees of the four regions/combinations were built separately. Samples of Brugmansia arborea were clustered into one clade, and the other species of Datura L. formed another clade. The results showed that either ITS2 or psbA-trnH was useful to identify Daturae Flos from its adulterants.
Base Sequence ; DNA Barcoding, Taxonomic ; methods ; DNA, Intergenic ; genetics ; DNA, Plant ; genetics ; Datura ; classification ; genetics ; Datura metel ; genetics ; Datura stramonium ; genetics ; Drug Contamination ; Flowers ; genetics ; Phylogeny ; Plants, Medicinal ; genetics ; Polymerase Chain Reaction ; Polymorphism, Single Nucleotide ; Sequence Analysis, DNA ; Solanaceae ; genetics ; Species Specificity

Angel's trumpet is a poisonous shrub recently imported to Korea for ornamental PURPOSE. It contains tropane alkaloids and the ingestion of any part of the plant can cause serious anticholinergic toxicity. Not only accidental ingestion, intentional intake as hallucinogen is not uncommon in the United States and Europe. We report a woman who ingested angel's trumpet by mistake and showed anticholinergic syndrome. The potential public medical harm of this widespread plant should be reported in the media, along with the associated dangers from misuse as a hallucinogen.
Alkaloids ; Datura ; Eating ; Europe ; Female ; Humans ; Korea ; Plants ; Solanaceous Alkaloids ; United States