平贝母药材质量标准研究

目的 对中药材平贝母的水分、灰分、酸不溶性灰分和贝母乙素含量等进行研究.方法 照2010年版<中国药典(一部)>中方法使用水分测定法测定水分,灰分测定法测定总灰分和酸不溶性灰分.采用高效液相色谱法测定贝母乙素含量,色谱柱为Symmetry C18柱(150 mm×3.9 mm,5μm),流动相为CH3CN-100 mmol/L CH3COONH4(43:57),流速为1.1 mL/min,检测波长为375 nm,柱温为室温,进样10μL.结果 中药材平贝母水分不应超过13%,总灰分不高于3.5%,酸不溶性灰分不得高于0.5%;贝母乙素含量测定平均回收率为98.13%,RSD=2.57%(n=6).结论 为建立和完善平贝母的质量标准体系提供了可供参考的依据.     

5种贝母类药材的定性鉴别

目的:考察浙贝母、川贝母、湖北贝母、平贝母和伊贝母的粉末显微鉴别特征及薄层色谱(TLC)鉴别特征。方法:按《中国药典》方法对5种贝母粉末进行显微鉴别;采用2种展开系统,分别对5种贝母进行TLC鉴别。结果:得到5种贝母粉末的显微特征区别点。浙贝母、川贝母、湖北贝母、平贝母的TLC图谱中均含有贝母素甲和贝母素乙,均无西贝母碱;伊贝母的TLC图谱中无贝母素甲和贝母素乙,但含有西贝母碱。结论:本试验结果可为5种贝母类药材的鉴别提供参考依据。     

安徽贝母中的新二萜成分

为了研究安徽贝母(Fritillaria anhuiensis S.C.Chen et S.E.Yin)的化学成分。将安徽贝母的干燥鳞茎用95%乙醇回流提取后，用硅胶柱色谱进行分离纯化，得到了3个化合物。根据化合物的理化性质和光谱数据进行结构鉴定，分别为12,15-epoxy-8(17),13-labdadien-19-ol (1)，ent-3β-acetoxy-kauran-16β,17-diol (2)，ent-kaurane-3β,16β,17-triol (3)。化合物1为新的labdane型二萜类化合物，化合物2和3为首次从该植物中得到。     

Differentiation-inducing effects of verticinone,an isosteroidal alkaloid isolated from the bulbus of Fritillaria ussuriensis,on human promyelocytic leukemia HL-60 cells

The inducer of differentiation of human promyelocytic leukemia HL-60 cells is commonly accepted to have potential therapeutic importance. Verticinone, one of the major isosteroidal alkaloids from the bulbus of Fritillaria ussuriensis, was found to inhibit the growth of HL-60 cells by inducing these cells to differentiate toward granulocytes. Importantly, the combination of verticinone with all-trans retinoic acid (ATRA), a well-known inducer of HL-60 cells into granulocytic lineages, was more effective than either alone, suggesting its therapeutic use in minimizing the effective dose of ATRA.     

从平贝母茎叶中分离的平贝啶甙的结构

本文报道自平贝母(Fritillaria ussuriensis Maxim)的地上茎叶中分得一种新的甾体生物碱甙,定名为平贝啶甙(pingbeidinoside),mp 242～243.2℃[a]_D²⁵+6.9°(c 0.145,MeOH)。经IR,MS,¹HNMR和¹³CNMR谱解析,推定其结构为3β,16α,20-三羟基-Δ⁵-22,26-缩亚胺胆甾烷-25-O-α-D-葡萄糖甙。     

HPLC-ELSD法同时测定平贝母中贝母素甲、贝母素乙的含量

目的：建立HPLC—ELSD法同时测定平贝母中贝母素甲、贝母素乙含量的方法,并对市售样品进行考察,制定含量限度。方法：色谱柱：Waters C18色谱柱（150mm×4．6mm,5μm）,流动相：乙睛-二乙胺-水（70：0．03：30）,流速：1．0ml·min^-1,ELSD参数：漂移管温度90℃,载气流速为2．2L·min^-1。结果：贝母素甲、贝母素乙分别在0．06～0．35μg和0．08—0．50μg范围内线性关系良好。平均回收率（n=6）分别为98．4％,98．9％,RSD分别为1．8％,1．9％。结论：此法简便,快捷,结果可靠,可用于平贝母药材的质量评价,为今后其质量标准的修订提供了依据。     

平贝碱丙的结构

本文报道自东北平贝母(Fritillaria ussuriensis Maxim)中分得一种新的C-去甲-D-异甾体生物碱,命名为平贝碱丙(pingbeimine C)。根据红外、质谱、氢谱、碳谱及X-射线衍射,确定其结构式为Ⅳ式。     

平贝母的化学成分研究——Ⅱ.平贝碱甙的分离和鉴定

A new C-nor-D-homosteroid alkaloid glucoside, pingpeimine glucoside, was isolated from Fritillaria ussuriensis Maxim. Based on preparation of derivative and spectral studies, the structure of pingpeimine glucoside was elucidated as Ⅰ.     

青羚散的薄层色谱鉴别

目的：研究青羚散的质量控制方法。方法：采用薄层色谱(TCL)鉴别对青羚散中大黄平贝母、黄芩进行鉴别。结果：三味药材的薄层色谱鉴别至阳性，阴性对照无干扰。结论：方法可靠对青羚散的质量控制有重要意义。     

HPLC法测定平贝母中腺苷的含量

目的：建立HPLC法测定平贝母中腺苷含量。方法：色谱柱：Inertsil ODS—SP（250mm×4．6mm,5μm）;流动相：水-乙腈（93：7）;流速：1．0ml·min^-1;柱温：室温;检测波长：260nm。结果：腺苷在0．04～0．24μg与峰面积线性关系良好（r=0．9999）,平均加样回收率为96．32％,RSD=1．15％（n=5）。结论：本方法简便、准确、重复性好,可用于平贝母的质量控制。     

Steroidal alkaloids from the bulbs of Fritillaria unibracteata

Two new steroidal alkaloids peimisine-3-O-β-D-glucopyranoside (1) and puqiedinone-3-O-β-D-glucopyranoside (3), together with three known compounds peimisine (2), puqiedinone (4), and puqiedine (5), were isolated and characterized from the bulbs of Fritillaria unibracteata. Their structures were fully elucidated by spectroscopic and chemical methods. Compound 1 showed moderate protection effect on neurotoxicity of PC12 cell lines induced by rotenone.     

Antioxidant constituents from the stem ofSorghum bicolor

The EtOAc soluble fraction from the stem of Sorghum bicolor showed a strong free radical scavenging activity. Five major compounds were isolated from this fraction. They were identified by spectral data as methyl ferulate (1), methyl p-hydroxycinnamate (2), p-hydroxybenzaldehyde (3), tricin (4), and quercetin 3,4'-dimethyl ether (5). Among these compounds, 1 exhibited a strong, free radical scavenging activity on 1,1-diphenyl-2-picrylhydrazyl (DPPH) with an IC50 value of 0.7 microM. We further studied the effects of these isolated compounds on the lipid peroxidation in rat liver microsomes induced by non-enzymatic method. All five compounds showed anti-lipid peroxidation activity (IC50 values of 0.5, 0.4, 0.3 and 0.3 microM, respectively).     

Neuroprotective and free radical scavenging activities of phenolic compounds fromHovenia dulcis

The EtOAc-soluble fraction from a methanolic extract of Hovenia dulcis Thunb. exhibited neuroprotective activity against glutamate-induced neurotoxicity in mouse hippocampal HT22 cells. The neuroprotective activity-guided isolation resulted in 8 phenolic compounds (1-8), such as vanillic acid (1), ferulic acid (2), 3,5-dihydroxystilbene (3), (+)-aromadendrin (4), methyl vanillate (5), (-)-catechin (6), 2,3,4-trihydrobenzoic acid (7), and (+)-afzelechin (8). Among these, compounds 6 and 8 had a neuroprotective effect on the glutamate-induced neurotoxicity in HT22 cells. Furthermore, compound 6 had a DPPH free radical scavenging effect with an IC50 value of 57.7 microM, and a superoxide anion radical scavenging effect with an IC50 value of 8.0 microM. Both compounds 6 and 8 had ABTS cation radical scavenging effects with IC50 values of 7.8 microM and 23.7 microM, respectively. These results suggest that compounds 6 and 8 could be neuroprotectants owing to their free radical scavenging activities.     

The influence of some 3-amino-2-pyrazoline derivatives on cyclooxygenase and lipoxidase activities

The influence of 3-amino-1-[m-(trifluoromethyl)-phenyl]-2-pyrazoline (BW 755C), 1-(3,3-diphenylpropyl)-3-amino-5-methyl-2-pyrazoline (KD 785) and 1-(3,3-diphenylpropyl)-3-amino-2-pyrazoline (KD 679) on cyclooxygenase and lipoxidase activities has been studied. All three compounds inhibited soybean and platelet lipoxidase activity. BW 755C was a much stronger inhibitor than KD compounds. The tested compounds stimulated oxygen consumption by cyclooxygenase from ram seminal vesicle microsomes in the presence of 100 microM of arachidonic acid in the range of concentrations between 30 and 300 microM and increased PGE2 generation in these experimental conditions. Radiochemical studies showed that BW 755C had been a stimulator of cyclooxygenase in higher (100 microM) substrate concentrations but, in agreement with a previous report it acted as an inhibitor of this enzyme when substrate concentration was 1.6 microM. Probably this component acted as a free radical scavenger.     

Spasmolytic effects of three harmala alkaloids on guinea-pig isolated trachea

The present study examined and compared the spasmolytic effects of 3 harmala alkaloids, harmine, harman, and harmaline, on carbachol-, histamine-, and KCl-induced contractions of guinea-pig isolated tracheal preparations. All 3 compounds relaxed the tracheal preparations contracted by these spasmogens with similar or different EC50 values, harmine being the most potent one. The cumulative concentration-response curves of all 3 compounds for carbachol-induced contraction were shifted to the right by propranolol (1 microM) pretreatment, indicating the involvement of the activation on the beta-adrenoceptors. All 3 compounds shifted the concentration-response curves of carbachol to the right in a parallel manner with the pA2 values comparable with their relaxation EC50 values, indicating a competitive antagonism at the muscarinic receptors. Receptor binding assays indicated that all 3 compounds interacted with lung muscarinic receptors (Ki = 11-13 microM), histamine H1 receptors (Ki = 27-107 microM), and beta2-adrenoceptors (Ki = 20-51 microM). Therefore, in addition to their actions on receptor-linked and voltage-dependent Ca2+ channels as reported in other types of smooth muscle, the present study suggests that the actions on muscarinic receptors, histamine H1 receptors, and beta2-adrenoceptors are also involved in their spasmolytic effects on airway smooth muscles.     

A potent anti-complementary acylated sterol glucoside fromOrostachys japonicus

In order to isolate substances that inhibit the hemolytic activity of human serum against erythrocytes, we have evaluated whole plants of the Orostachys japonicus species with regard to its anti-complement activity, and have identified its active principles following activity-guided isolation. A methanol extract of the O. japonicus, as well as its n-hexane soluble fraction, exhibited significant anti-complement activity on the complement system, which was expressed as total hemolytic activity. A bioassay-guided chromatographic separation of the constituents resulted in the isolation of three known compounds 1-3 from the active n-hexane fraction. The structure of these compounds were analyzed, and they were identified as hydroxyhopanone (1), beta-sitosteryl-3-O-beta-D-glucopyranosyl-6'-O-palmitate (2), and beta-sitosteryl-3-O-beta-D-glucopyranoside (3), respectively. Of these compounds, compound 2 exhibited potent anti-complement activity (IC50= 1.0 +/- 0.1 microM) on the classical pathway of the complement, as compared to tiliroside (IC50= 76.5 +/- 1.1 microM), which was used as a positive control. However, compounds 1 and 3 exhibited no activity in this system.     

Steroids from Saussurea ussuriensis

One novel and seven known steroids were isolated from the alcohol extract of the whole plant of Saussurea ussuriensis. The structure of the new compound was elucidated as stigmast-5-en-3beta,7alpha,22-triol (1) by spectroscopic methods including intensive 2D NMR techniques (H1-H1 COSY, gHSQC, gHMBC) and HR-SI-MS and the known compounds were identified on the basis of comparing their NMR data with those of corresponding compounds in the literature.     

Comparative efficacy of three 2'-fluoropyrimidine nucleosides and 9-(1,3-dihydroxy-2-propoxymethyl)guanine (BW B759U) against pseudorabies and equine rhinopneumonitis virus infection in vitro and in laboratory animals

The three 2'-fluoropyrimidine nucleosides 1-(2-deoxy-2-fluoro-beta-D-arabinofuranosyl)-5-iodocytosine (FIAC), 1-(2-deoxy-2-fluoro-beta-D-arabinofuranosyl)-5-iodouracil (FIAU), and 1-(2-deoxy-2-fluoro-beta-D-arabinofuranosyl)-5-methyluracil (FMAU), showed high activity in RK13 monolayers against equine rhinopneumonitis virus, (EHV-1, IC50 range 0.02-0.18 microM), Aujeszky's disease virus (SHV-1, pseudorabies, IC50 range 0.25-7 microM) and infectious bovine rhinotracheitis virus (1BR, BHV-1, IC50 range 0.1-3 microM). The activity of these compounds was compared with 9-(1,3-dihydroxy-2-propoxymethyl)guanine (BW B759U, DHPG) in two laboratory animal disease models: EHV-1-induced hepatitis in hamsters and SHV-1-induced encephalitis in mice. All the compounds, provided from 3 to 5 h pre-infection for 5 days, were effective in preventing EHV-1 mortality (at 3-5 mg/kg per day) and in significantly reducing SHV-1 mortality (at 60 mg/kg per day). While FIAU had the greatest activity in vitro, FMAU tended to be more potent in vivo. The reasons for these differences between relative in vitro and in vivo activities are briefly discussed.     

Mediation of nitric oxide from photosensitive stores in the photorelaxation of the rabbit corpus cavernosum

Isolated rabbit corpus cavernosum relaxed in response to ultraviolet (UV) light (365 nm). The UV light-induced relaxation (photorelaxation) was diminished on repeated UV irradiation from 30.5+/-4.0% (the first photorelaxation) to 15.5+/-2.7% (the last photorelaxation). Hydroxocobolamine of 100 microM and hemoglobin (Hb) of 10 microM, which are nitric oxide (NO) scavengers, and 10 microM 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one (ODQ), a soluble guanylyl cyclase inhibitor, markedly reduced photorelaxation. However, 300 microM 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl 3-oxide (carboxy-PTIO) failed to inhibit photorelaxation. NaNO(2) and N(G)-nitro-L-arginine (L-NA) but not 3-nitro-L-tyrosine (3-NT) were found to be photosensitive in that these compounds are photolysed to release NO, as demonstrated by use of an amperometric NO probe; NO signals produced by 500 microM NaNO(2), and 500 microM L-NA were 133.3+/-28.9 and 54.4+/-10.4 pA, respectively. Not 3-NT but the other compounds (all 200 microM) also enhanced photorelaxation of the cavernosal tissue. Based on these findings, the substance, which mediates photorelaxation, could be NO released from putative stores in the rabbit corpus cavernosum, and L-NA as well as NaNO(2) but not 3-NT produce NO under the influence of UV light.