Discovery, isolation and structural identification of alkaloid glycosides in six traditional Chinese medicine such as Coptis chinensis / 中国中药杂志

Alkaloids are important active ingredients occurring in many traditional Chinese medicines, and alkaloid glycosides are one of their existence forms. The introduction of saccharide units improves the water solubility of alkaloid glycosides thus presenting better biological activity.Because of the low content in plants, alkaloid glycosides have been not comprehensively studied. In this study, ultrahigh performance liquid chromatography-quadrupole time of flight-tandem mass spectrometry(UPLC-QTOF-MS/MS) was employed to identify and analyze the alkaloid glycosides in Coptis chinensis, Phellodendron chinense, Menispermum dauricum, Sinomenium acutum, Tinospora sagittata and Stephania tetrandra. The results showed that except Tinospora sagittata, the other five herbal medicines contained alkaloid glycosides. Furthermore, the alkaloid glycosides in each herbal medicine were identified based on UV absorption spectra, quasimolecular ion peaks in MS, fragment ions information in the MS/MS, and previous literature reports. A total of 42 alkaloid glycosides were identified. More alkaloid glycosides were identified in C. chinensis and Menispermum dauricum, and eleven in C. chinensis were potential new compounds. Furthermore, the alkaloid glycosides in the water extract of C. chinensis were coarsely se-parated by macroporous adsorption resin, purified by column chromatography with D151 cation exchange resin, ODS and MCI, combined with semi-preparative high performance liquid chromatography. Two new alkaloid glycosides were obtained, and their structures were identified by mass spectrometry and NMR data as(S)-7-hydroxy-1-(p-hydroxybenzyl)-2,2-N,N-dimethyl-1,2,3,4-tetrahydroisoquinoline-6-O-β-D-glucopyranoside and(S)-N-methyltetrahydropalmatubine-9-O-β-D-glucopyranoside, respectively. This study is of great significance for enriching the information about the chemical composition and the in-depth development of C. chinensis. Meanwhile, it can provide a reference for rapid identification and isolation of alkaloid glycosides from other Chinese herbal medicines.

Chemical constituents from rhizome of Menispermum dauricum and their anti-hypoxic activities / 中国中药杂志

To study the chemical constituents from the rhizome of Menispermum dauricum,fifteen compounds,N-methylcorydaldine( 1),thalifoline( 2),stepholidine( 3),acutumine( 4),daurisoline( 5),acutumidine( 6),dauricicoline( 7),bianfugecine( 8),6-O-demethylmenisporphine( 9),bianfugedine( 10),dauricoside( 11),eleutheroside D( 12),aristolactone( 13),aristoloterpenateⅠ( 14) and aristolochic acid( 15) were isolated from 75% ethanol extract of Menispermi Rhizoma by using thin layer chromatography and column chromatography methods. Their structures were identified based on their physicochemical properties and spectral data. Among them,compounds 12-15 were obtained from the genus Menispermum for the first time. Six alkaloids with higher contents were subjected to evaluate the anti-hypoxic activities by using MTT method. As a result,six alkaloids exhibited significant anti-hypoxia activities.

Effect of Phenolic Alkaloids from Menispermum dauricum on EAAC1 mRNA Expression in Hippocampal Neurons of Rats with Focal Cerebral Ischemia / 医药导报

Objective To investigate the effect of phenolic alkaloids from Menispermum dauricum( PAMd)on the mRNA expression of the glutamate transporter EAAC1 in hippocampal neurons of rats subjected to focal cerebral ischemia and to elucidate its neuroprotective mechanisms. Methods A total of 42 Sprague Dawley(SD)rats were randomly divided into three groups:sham group,model control group,and PAMd(10 mg·kg-1 ,i.g)group(n = 14 each).The focal cerebral ischemia model of rat was induced by the middle cerebral artery occlusion( MCAO). The 2,3,5-triphenyltetrazolium chloride( TTC) staining was applied to measure the cerebral infarct size and the reverse transcriptase-polymerase chain reaction(RT-PCR)assay to detect EAAC1 mRNA expression in hippocampal neurons. Results After 24 h ischemia,the cerebral infarct volumes were (0.0±0.0)%,(35.3±2.9)% and(21.3±3.8)% in sham group,model control group and PAMd group,respectively(P<0.05). The relative expression levels of EAAC1 mRNA were 0.97±0.04,2.46±0.13,and 1.91±0.15 in the three groups,respectively (P<0. 05). Conclusion PAMd may protect against cerebral ischemia by up-regulating EAAC1 mRNA expression and alleviating the excitotoxicity of glutamic acid.

Identification on pathogen of Menispermum dauricum target spot and study on its biological characteristics and fungicide susceptibility / 中草药

Objective: To identify the pathogen of Menispermum dauricum target spot (a new plant disease in China) and study its biological characteristics and susceptibility to fungicides. Methods: Tissue isolation method was used to obtain the isolates from diseased-leaf. The pathoginicity of the isolates was fulfilled according to Koch's postulate. The identification of the pathogen was carried out according to the morphological and cultural characteristics and rDNA-ITS sequence analysis. The effects of culture medium, temperature, pH value, carbon and nitrogen sources, and light on mycelium growth and sclerotia production of the pathogen were studied. The mycelium growth rate was used to test the susceptibility of pathogen for 14 fungicides. Results: The M. dauricum target spot was caused by Streptobotrys caulophylli. The optimal medium for mycelium growth was PDA; PDA and PSA media were suitable for the sclerotium production. The optimal temperature ranges for mycelium growth and spore production were 20-28℃ and 10-30℃ and were suitable for sclerotium production. The suitable pH values for mycelium growth and sclerotium production were 4-9 and 5-11, respectively. Sucrose and L-glutamine were the optimal carbon and nitrogen sources for mycelium growth. Synanthrin and sucrose were the optimal carbon source and sodium nitrate was nitrogen source for sclerotium production. The total light could promote mycelium growth, while the darkness could promote sclerotium production. The pathogen was sensitive to procymidone, cyprodinil, iprodione, fludioxonil, metalaxyl•mancozeb, and pyraclostrobin•metiram with EC50 < 1.0 mg/L and EC90 < 5.0 mg/L. Conclusion: It is the first report on M. dauricum target spot caused by S. caulophylli in China. The suitable conditions (culture medium, temperature, pH value, carbon and nitrogen sources, and light) for mycilium growth and sclerotium production are determined. The above six fungicides are screened as further field trial agents for disease control.

Inhibitory Effect of Phenolicalkaloids of Menispermum Dauricum on the BXPC-3 Tumor and Its Effect on iNOS Content in Tumor-bearing Mice / 中国药房

OBJECTIVE:To observe the inhibitory effect of phenolicalkaloids of menispermum dauricum(PAMD)on the human pancreatic cell line(BXPC-3)tumor and study the effect of PAMD on serum content of inducible nitric oxide synthase(iNOS)in tumor-bearing mice.METHODS:The BXPC-3 tumor-bearing mice model was established,and the model mice were randomly assigned to 6 groups:model control group,blank control group,cyclophosphamide group,3 PAMD groups(high,medium and low dosages).The blank control group was not inoculated with tumor strain but given same volume of normal saline.The tumor-inhibition ratio of PAMD was detected and the content of iNOS of in peripheral blood of tumor-bearing mice was determined by fluorospectrophotometry.RESULTS:PAMD(high,medium and low dosages)showed marked inhibitory effect on tumor growth of BXPC-3 cell line in tumor-bearing mice,with the inhibition ratios at 34.91%,52.83% and 41.51%,respectively.As compared with model control group,PAMD-treated groups showed significantly lower content of iNOS in peripheral blood of tumor-bearing mice.CONCLUSION:PAMD showed marked inhibitory effect on tumor growth of BXPC-3 cell line in tumor-bearing mice,which might be attributed to the lowered content of iNOS.