The Relationship Between Neuroprotective Activity and Antigenotoxic and Acetylcholinesterase Inhibitory Effects of Glaucium corniculatum Extracts

Abstract The purpose of this study was to investigate the relationship between the acetylcholinesterase (AChE) inhibitory and antigenotoxic effect with the neuroprotective activity of Glaucium corniculatum methanol and water extracts rich in rutin and quercetin flavonoids. Neuroprotective activity in terms of cell survival and development against oxidative damage was measured by MTT assay and microscopic analysis in H2O2-induced NGF-differentiated PC12 (dPC12) cells. QRT-PCR and western blot hybridization method was employed for the determination of AChE inhibition of the extracts in the same cell model, and the genotoxic and antigenotoxic effects were identified with Comet assay with human lymphocytes. H2O2-induced vitality loss in dPC12 cells was inhibited in pre-treated cells with these plant extracts. Moreover, extracts stimulated neurite formation and prevented the oxidative stress-induced reduction in neurite growth. In general, it was determined that G. corniculatum methanol extract containing higher amounts of rutin and quercetin was more effective than water extract in terms of AChE inhibitory, antigenotoxic and also neuroprotective effect. In this study, it was shown for the first time that both AChE inhibitory and antigenotoxic effects of G. corniculatum may be effective in neuroprotection and it's protective and therapeutic effects against neurodegeneration may be related to the flavonoid content.

Chemical profile, acetylcholinesterase, butyrylcholinesterase, and prolyl oligopeptidase inhibitory activity of Glaucium corniculatum subsp. refractum

Abstract Papaveraceae is one of the prominent alkaloid-containing families, and plants of the genus Glaucium (Papaveraceae) are known for their bioactive alkaloids. Glaucium species have been used in traditional medicine in Turkey as an analgesic, narcotic, sedative, and antitussive. In this study, it was planned to evaluate the inhibitory activity of an alkaloidal extract of Glaucium corniculatum subsp. refractum on acetylcholinesterase (AChE), butyrylcholinesterase (BuChE) and prolyl oligopeptidase (POP), as well as exploring the chemical profile of the plant by using Gas Chromatography-Mass Spectrometry (GC-MS). The AChE, BuChE and POP inhibition activities of the alkaloidal extract of G. corniculatum subsp. refractum were determined spectrophotometrically. A rapid GC-MS method was used to identify alkaloids that could be responsible for these inhibition activities. In total, eleven alkaloids were identified in the alkaloid extract of the plant by GC-MS. Allocyptopine (52.92%) and protopine (25.38%) were found as the major constituents. The alkaloidal extract of G. corniculatum subsp. refractum showed potent AChE inhibitory activity (IC50:1.25 µg/mL) and BuChE inhibitory activity (IC50: 7.02 µg/mL). The extract also showed a remarkable inhibitory effect on POP with an IC50 value of 123.69 µg/mL. This study presents the first GC-MS investigation and POP inhibitory activity of G. corniculatum subsp. refractum.

Alkaloids from Macleaya cordata and their cytotoxicity assay / 中国中药杂志

<p><b>OBJECTIVE</b>To study the alkaloids of Macleaya cordata and their anti-tumor activities.</p><p><b>METHOD</b>Alcohol and liquid-liquid extraction were used methods were used to extract the alkaloids constituents, and silica gel, reverse-phase octadecylsilyl (ODS), sephadex LH-20 chromatographic methods and HPLC were applied to isolate and purify compounds. MS, NMR spectroscopic methods were used to determine their structures. Furthermore, the cytotoxicity of these chemical components for MCF-7 and SF-268 cell lines was measured by MTT method.</p><p><b>RESULT</b>Twelve alkaloids were isolated from the fruits of M. cordata, and their structures were identified as: maclekarpine E (1), 6-acetonyldihyrochelerythrine (2), cavidilinine (3), 6-acetonyldihyrosanguinnarine (4), O-methylzanthoxyline (5), 6-methoxy-dihydrosanguinarine (6), spallidamine (7), 6-hydroxyldihydrochelerythrine (8), arnotianamida (9), dihydrosanguinarine (10), protopine (11), and cryptopine (12).</p><p><b>CONCLUSION</b>Compounds 1, 3, 7-9 were isolated from M. cordata for the first time, and compound 5 is a new natural product. The results of cytotoxic assay indicated that compound 6 showed strong cytotoxicity against MCF-7 and SF-268 cell lines with IC50 values of 0.61 μmol · L(-1) and 0.54 μmol · L(-1), respectively.</p>

Chemical constituents of the roots of Macleaya microcarpa and activation efficacy of benzophenanthridine alkaloids for the transcription of xbp1 gene / 药学学报

Ongoing study on the chemical constituents of the roots of Macleaya microcarpa led to the isolation of eight compounds of derivatives of triterpenes and organic acids in addition to some previously identified benzophenanthridines. The eight compounds were identified by spectroscopic methods as well as comparison with literature values as 1-oxo-2, 22 (30)-hopandien-29-oic acid (1), 3-oxo-12-oleanen-30-oic acid (2), 3α-hydroxy-12-oleanen-30-oic acid (3), 3β-hydroxy-12-oleanen-30-oic acid (4), ferulic acid (5), ferulic acid 4-O-β-D-glucoside (6), 3-O-feruloylquinic acid (7), and methyl 3-O-feruloylquinate (8). Of which, 1 is a new triterpenoid of hopanes and 2-8 are isolated from M microcarpa for the first time. In order to discover natural active compounds as potential agents of anti-ulcerative colitis (UC), an in vitro drug high-throughput screening model targeted x-box-binding protein 1 (xbp1) was employed to evaluate the activity of the major chemical constituents of M microcarpa. The result confirmed that two dihydrobenzophenanthridines, dihydrosanguinarine (9) and dihydrochelerythrine (10), showed a certain activity on activating the transcription of xbpl, a transcription factor (TF) associated with the occurrence, development, and potential treatment of UC, with their relative activating ratios being 1.76 and 1.77 times, respectively, as compared with control group.

Identification and analysis of Corydalis boweri, Meconopsis horridula and their close related species of the same genus by using ITS2 DNA barcode / 中国中药杂志

The study is aimed to ensure the quality and safety of medicinal plants by using ITS2 DNA barcode technology to identify Corydalis boweri, Meconopsis horridula and their close related species. The DNA of 13 herb samples including C. boweri and M. horridula from Lhasa of Tibet was extracted, ITS PCR were amplified and sequenced. Both assembled and web downloaded 71 ITS2 sequences were removed of 5. 8S and 28S. Multiple sequence alignment was completed and the intraspecific and interspecific genetic distances were calculated by MEGA 5.0, while the neighbor-joining phylogenetic trees were constructed. We also predicted the ITS2 secondary structure of C. boweri, M. horridula and their close related species. The results showed that ITS2 as DNA barcode was able to identify C. boweri, M. horridula as well as well as their close related species effectively. The established based on ITS2 barcode method provides the regular and safe detection technology for identification of C. boweri, M. horridula and their close related species, adulterants and counterfeits, in order to ensure their quality control, safe medication, reasonable development and utilization.

Chemical constituents from a Tibetan medicine Meconopsis horridula / 中国中药杂志

A phytochemical investigation on the aerial parts of a Tibetan medicine Meconopsis horridula, by solvent extraction, repeated chromatographies on silica gel, Sephadex LH-20, and preparative TLC techniques, led to the isolation of 9 compounds. By spectroscopic analysis and comparison of its 1H and 13C-NMR data with those in literatures, their structures were identified as oleracein E(1), N-( trans-p-coumaroyl) tyramine (2), chrysoeriol (3), apigenin (4), hydnocarpin (5), p-coumaric acid glucosyl ester (6), stigmast-5-ene-3beta-ylformate (7), 3beta-hydroxy-7alpha-ethoxy-24beta-ethylcholest-5-ene (8), and beta-sitosterol (9), respectively, among which compounds 6-8 were isolated from the genus for the first time,and 1,3 were isolated from the species for the first time. A MTT method was applied to evaluate the cytotoxic activity of compounds 14 against the human hepatocellular liver carcinoma cell line (HepG2), and compound 1 showed significant cytotoxicity against HepG2,with its inhibitory rate of 52.2% at 10 micromol x L(-1).

Isolation of endophytic fungi from Macleaya cordata and screening of sanguinarine-producing strains / 中国中药杂志

Endophytic fungi were isolated from Macleaya cordata growing in Dabie Mountain by agar-block method, and then the endophytic fungi were grouped into different types based on their morphological characteristics, and thin layer chromatography (TLC) and high performance liquid chromatography (HPLC) were employed to determine whether the metabolic substances contained sanguinarine or not, and then preliminarily identified by morphological method. The results showed that the leaves hosted the largest number of endophytes (96 isolates) followed by the stems (57 isolates) and finally the roots (28 isolates), respectively. Based on morphological characteristics the endophytic fungi were grouped into 26 types in our study. TLC and HPLC results showed that there was sanguinarine in the metabolic substances of BLH 51 strain. According to the morphological characteristic, the BLH 51 strain was identified as Fusarium proliferatum. All these indicated that the medicinal plant M. cordata harbors abundant endophytes, which could be a new source for the search of active secondary metabolites.

Study on material base of corydalis rhizoma / 中国中药杂志

Corydalis Rhizoma, the dried tuber of Corydalis yanhusuo (Papaveraceae) distributed traditionally mainly in south-eastern and now cultivated in northwestern and other district in China, is one of the commonly used and well-known traditional Chinese medicine. It has been widely used to treat spastic pain, abdominal pain, pain due to injury, and promote blood circulation. Its main chemical constituents were alkaloids, which were divided into the two types of protoberberines and aporphines. Among them, some alkaloids were found to elicit profound effects on the dopaminergic system in the central nervous system, which plays an important role in regulating nociception. In this article, the chemical composition and structure-types, new methods of qualitative and quantitative analysis as well as characteristics of biotransformation, absorption, distribution, metabolism, excretion, pharmacokinetic, and drug-drug interaction for the alkaloids were revealed. These results would greatly contribute to the establishment of bioactive material base of Corydalis Rhizoma.

Chemical components of essential oils from Meconopsis oliverana and their antioxidant activity / 中国中药杂志

<p><b>OBJECTIVE</b>To study the chemical components of essential oils from Meconopsis oliverana and their antioxidant activity.</p><p><b>METHOD</b>The essential oil was extracted by steam distillation, and GC-MS analysis was used to identify its constituents. The OH free radical scavenging activity of the essential oils was evaluated with an enzyme mark instrument by assay of the ability of DPPH free radical scavenging. BHT was used as positive control.</p><p><b>RESULT</b>Forty-seven compounds, account for 91.866% of the essential oils, were identified. The ability of scavenging OH and DPPH radicals of the essential oils is stronger than that of BHT.</p><p><b>CONCLUSION</b>The main chemical constituents of the essential oils from M. oliverana are n-hexadecanoic acid (27.653%) and 6,10,14-trimethyl-2-pentadecanone (16.330%). And the essential oils showed strong antioxidant activity.</p>

Etude comparee des effets anti-hepatotoxiques d'extraits d'Argemone mexicana L. (Papaveraceae); une plante utilisee dans le traitement traditionnel de la jaunisse au Burkina Faso

Argemone mexicana L. (Papaveraceae) fait l'objet de nombreuses indications therapeutiques en medecine traditionnelle dans la region des Cascades au Burkina Faso. Dans une etude preliminaire de 2006; il a ete mis en evidence une activite anti-hepatotoxique du decocte aqueux lyophilise des feuilles chez le rat Wistar prealablement icterise par une administration de tetrachlorure de carbone CCl4 (0;5 mL/kg; i.p.). Dans la presente etude; les auteurs projettent de comparer les effets anti-hepatotoxiques de trois (03) extraits de la plante notamment deux extraits aqueux lyophilises (decocte et macere) et un extrait du totum alcaloidique; tous administres a differents lots de rats rendus initialement icteriques par injection de l'hepatotoxique. La silymarine (100 mg/kg) a ete utilisee comme produit de reference anti-hepatotoxique. Les trois extraits (250 mg/kg; p.o.) ont temoigne une action anti-hepatotoxique significative (p0;05) objectivee par un pourcentage de reduction notable des marqueurs biochimiques enzymatiques notamment les transaminases (ASAT/GOT; ALAT/GPT); la phosphatase alcaline (PAL) et la bilirubine directe (DBil). Les pourcentages moyens de reduction d'hepato-toxicite induite ont ete de 79;4 (silymarine); 69;73 (decocte lyophilise); 70;63 (macere lyophilise) et 72 (totum alcaloidique). Ces resultats illustrent d'une part la preeminence de l'action pharmacologique anti-hepatotoxique de l'extrait de totum alcaloidique compare aux extraits aqueux lyophilises et d'autre part; l'effet des extraits pris isolement reste en deca de celui de la silymarine

Study on semi-synthetic transforming technology for the natural product of isocorydione / 药学学报

Transforming technology for semi-synthesized isocorydione from the natural product ofisocorydine was studied. The factors affecting on the reaction yield were investigated. UV spectrophotometry was used to indicate the semi-synthesized yield of isocorydione. The optimum reaction conditions were determined as following: reacting for 12 h in the solution of sodium dihydrogen phosphate at pH 10, the temperature was 25 degrees C and the ratio of isocorydine to Fremy's radical was 1 : 2. Under the optimum conditions, the yield could reach up to 50.0%. The molecular structure of isocorydione was elucidated by X-ray single-crystal diffraction analysis for the first time.

Determination of o-methylflavinantine in a Tibetan medicine Meconopsis quintuplinervia by HPLC / 中国中药杂志

<p><b>OBJECTIVE</b>To develop an HPLC method for the determination of a Tibetan medicine Meconopsis quintuplinervia.</p><p><b>METHOD</b>A Hypersil-Keystone-C18 column (4.6 mm x 250 mm, 5 microm) was used with the isocratic elution of acetonitrile and 0.012% glacial acetic acid. The flow rate was 1.0 mL x min(-1), and the detection wavelength was set at 237 nm.</p><p><b>RESULT</b>The linear range of 0-methylflavinantine was 0.2-2.4 microg (r = 0.999 7). The average recovery was 96.26%.</p><p><b>CONCLUSION</b>The developed method was reliable, and can be used for the quality control of M. quintuplinervia Regel.</p>

Screen for natural benzylisoquinoline alkaloids against tumor / 中国中药杂志

<p><b>OBJECTIVE</b>To search for natural benzylisoquinoline alkaloids against tumor.</p><p><b>METHOD</b>In this study, taken DNA as target, a method was developed for screening of natural benzylisoquinoline alkaloids against tumor from traditional Chinese medicine by the use of centrifugal ultrafiltration combined with HPLC technology. The anti-tumor activity for the compounds screened was evaluated in vitro.</p><p><b>RESULT</b>Nine compounds interacted with DNA were discovered and identified from Macleaya cordata, Chelidonium majus, Coptis chinensis, and the proliferation of four types of human solid cancer cell lines was markedly inhibited by these compounds.</p><p><b>CONCLUSION</b>The developed method was considered to be suitable for screening of natural benzylisoquinoline alkaloids against tumor from traditional Chinese medicine.</p>

Expression of Bcl-2 and Bax proteins in myocardial cells after Macleaya cordata alkaloids poisoning in rats / 法医学杂志

OBJECTIVE@#To observe the expression of Bcl-2 and Bax proteins in rat's myocardial cells after Macleaya cordata alkaloids poisoning, and to provide certain molecular biology references for the detection of Macleaya cordata alkaloids poisoning.@*METHODS@#Experimental model of Macleaya cordata alkaloids poisoning was established, the expression levels of Bcl-2 and Bax proteins in these cells were detected by immunohistochemistry, and the results were analyzed by computer image system.@*RESULTS@#The expression levels of Bcl-2 and Bax proteins in myocardial cells in poisoning groups were much greater than those in the control groups (P<0.05).@*CONCLUSION@#If the clinical symptoms may not be obvious, the detection of Bcl-2 and Bax proteins level by immunohistochemistry still could be ancillary method.

Alkaloids from Macleaya cordata / 中国中药杂志

<p><b>OBJECTIVE</b>To study the alkaloids of Macleaya cordata.</p><p><b>METHOD</b>The alkaloids were isolated and purified by various column chromatography as well as recrystallization. Their structures were identified by physico-chemical properties and spectral analysis.</p><p><b>RESULT</b>Thirteen alkaloids were isolated and identified as 6-methoxy-dihydrosanguinarine (1), norsanguinarine (2), 6-acetonyl-dihyrochelerythrine (3), 6-acetonyl-dihyrosanguinnarine (4), sanguidimerine (5), chelidimerine (6), (+/-) -bocconarborine A (7), (+/-)-bocconarborine B (8), cryptopine (9), dihydrosanguinarine (10), dihydrochelerythrine (11), protopine (12), alpha-allocryptopine (13).</p><p><b>CONCLUSION</b>Compounds 2, 3, 5 - 9 were firstly isolated from the genus Macleaya and compound 1 was obtained from M. cordata for the first time.</p>

Isolation of functional total RNA from Argemone mexicana tissues

RNA extraction from recalcitrant plant tissues is frequently complicated by the presence of secondary metabolites, polysaccharides and polyphenols. These compounds may co precipitate with RNA, often rendering it unsuitable for either cDNA synthesis or hybridization in northern blot analyses and therefore, interfering with the gene analysis expression in such tissues. We have developed an efficient RNA extraction method from A. mexicana tissues. The procedure includes the use of polyvinylpolypyrrolidone (PVPP), to remove secondary metabolites, proteins and polyphenols, and a two-step precipitation with LiCl, to eliminate polysaccharides, and thus increasing RNA yield. The quality of the resulting RNA was evaluated spectrophotometrically and by agarose gel electrophoresis. Moreover, the RNA obtained by this method, could be used directly for both RT-PCR and northern blot analysis, without any further purification.

Impact of cytomixis on meiosis, pollen viability and pollen size in wild populations of Himalayan poppy (Meconopsis aculeata Royle).

We report the occurrence of cytomixis in wild populations of Himalayan poppy (Meconopsis aculeata Royle),which is considered to be an important and threatened medicinal plant growing in the high hills of the Himalayas. The impact of cytomixis on meiotic behaviour, reduced pollen viability and heterogeneous-sized pollen grains was also studied. Cytological studies in the seven wild populations from the high hills of Himachal Pradesh revealed that all the Himalayan populations exist uniformly at the tetraploid level (2n=56) on x=14. The phenomenon of chromatin transfer among the proximate pollen mother cells (PMCs) in six populations caused various meiotic abnormalities. Chromatin transfer also resulted in the formation of coenocytes, aneuploid, polyploid and anucleated PMCs. Among individuals that showed chromatin transfer, chromosome stickiness and interbivalent connections were frequently observed in some PMCs. The phenomenon of cytomixis in the species seems to be directly under genetic control; it affects the meiotic course considerably and results in reduced pollen viability.

Apoptosis in myocardial cells after macleaya cordata total alkaloids poisoning in rats / 法医学杂志

OBJECTIVE@#To observe pathological changes and apoptosis in rats myocardial cells after Macleaya cordata total alkaloids poisoning, and to provide some references for Macleaya cordata total alkaloids poisoning detection.@*METHODS@#An experimental model of Macleaya cordata total alkaloids poisoning was established, and the technology of TUNEL staining was used.The results were analyzed by computer image analysis competitive system.@*RESULTS@#Quantities of apoptosis in myocardial cells in poisoning groups were much more than those in the control groups at different tages (P<0.01). In addition the quantities of apoptosis were different after different poisoning duration.@*CONCLUSION@#Although clinical symptoms was not obvious and could not be detected by poison analysis. Pathological changes induced by Macleaya cordata total alkaloids could be found through the apoptosis detection.

Non-alkaloid constituents from a Tibetan medicine Meconopsis quintuplinervia / 中国中药杂志

<p><b>OBJECTIVE</b>To study the chemical constituents of a Tibetan medicine Meconopsis quintuplinervia.</p><p><b>METHOD</b>Column chromatographic techniques were applied to isolate constituents. A combination of IR, MS and NMR spectroscopy was used to identify structures of constituents.</p><p><b>RESULT</b>Twelve compounds were isolated from the ethanolic extract and their structures were elucidated as quercetin 3-O-beta-D-glucopyranoside (I), quercetin 3-O-beta-D-galactopyranosyl-(1-->6)-glucopyranoside (II), kaempferol 3-O-beta-D-glucopyranosyl-(1-->2)-beta-D-glucopyranoside (III), isorhamnetin 3-0-beta-D-galactopyranosyl-(1-->6)-beta-D-glucopyranoside (IV), caffeic acid (V), protocatechuic acid (VI), p-hydroxycinnamic (VII), 2-(3,4-dihydroxyphenyl )-ethyl-O-beta-D-glucopyranoside (VIII), p-hydroxybenzoyl-beta-D-glucopyranoside (IX), 4-O-beta-D-glucopyranosyl-(Z)-p-coumaric acid (X), 5, 7-dihydroxy-4H-4-chromenone (XI), daucosterol (XII).</p><p><b>CONCLUSION</b>Ten compounds were isolated from this genus for the first time except for XI and XII.</p>

Determination of protopine and isocorydine in root of Dactylicapnos scandens by HPLC / 中国中药杂志

<p><b>OBJECTIVE</b>To establish a HPLC method for determination of protopine and isocorydine in root of Dactylicapnos scandens.</p><p><b>METHOD</b>The separation was performed in a PHENOMENEX-C18 column with a mobile phase of methanol-0.2% phosphoric acid (adjusted to pH 7.0 with triethylamine)(50:50), The detection wavelength was at 254 nm and the flow rate was 0.8 mL x min(-1).</p><p><b>RESULT</b>The average recovery of Protopine and Isocorydine was 97.9%, 98.6% respectively, and RSD 1.3%, 1.4%.</p><p><b>CONCLUSION</b>This method is accurate, simple and reliable. It can be used for quality control of D. scandens.</p>