HPLC法测定独一味根中环烯醚萜苷和苯乙醇苷

目的建立HPLC法,同时测定独一味根中4种环烯醚萜苷胡麻属苷、山栀苷甲酯、7,8-dehydropenstemoside、8-O-乙酰山栀苷甲酯和2种苯乙醇苷连翘酯苷、毛蕊花糖苷。方法采用高效液相色谱法,Waters Symmetry柱(100 mm×3.5 mm,3.5μm),柱温25℃,流动相为甲醇-水,梯度洗脱;体积流量1.0 mL/min,检测波长235、332 nm。结果胡麻属苷在0.236～1.410μg、山栀苷甲酯在0.440～2.640μg、7,8-dehydropenstemoside在0.094～0.564μg、8-O-乙酰山栀苷甲酯在0.750～4.500μg、连翘酯苷在0.698～4.180μg、毛蕊花糖苷在0.455～2.730μg线性关系良好,回收率均在96.75%～103.97%,RSD均小于2%。结论此方法同时测定了独一味根中6种化学成分,方法分离度好,快速,简便,结果稳定。     

环烯醚萜苷类生物合成途径及相关酶的研究进展

环烯醚萜苷类化合物种类繁多,近年来对此类化合物的广泛研究发现其具有多种生物活性,如抗炎、抗肿瘤、保肝、心血管系统保护作用等。结合近年来国内外的研究,将其生物合成途径、相关酶(GPPS、GES、G10H、10-HGO、IS、7-DLS、7-DLGT、7-DLH、LAMT及SLS)及编码基因的研究进展进行综述,一方面可达到人为调节其代谢产物生成的目的,另一方面可为基于环烯醚萜苷类的新药发现提供参考。     

环烯醚萜苷类化合物体内代谢及代谢物药理活性研究进展

环烯醚萜苷类化合物为一大类广泛存在于中药中的具有多种药理活性的天然产物。在生物转化作用下,环烯醚萜苷类化合物体内主要以原型、苷元及Ⅰ,Ⅱ相代谢产物形式存在,其代谢产物已被证实具有多种药理活性。该文综述常见的几种环烯醚萜类化合物体内代谢及代谢物活性的研究进展,为进一步合理开发利用环烯醚萜类成分及其代谢产物提供一定的理论依据。     

不同产地和生长年限对巴戟天中环烯醚萜苷类成分形成和积累的影响

目的 探讨不同产地和生长年限对巴戟天Morinda officinalis中环烯醚萜苷类成分形成和积累的影响。方法 以HPLC法分析巴戟天环烯醚萜苷类成分的含量。色谱柱为艾杰尔Venusil MP C₁₈色谱柱（250 mm×4.6 mm,5 µm）;以0.2%磷酸＋0.01 mol/L磷酸氢二钠缓冲盐（A）-乙腈（B）为流动相,梯度洗脱;体积流量1.0 mL/min,柱温25 ℃;二极管阵列检测器（DAD）检测。结果 巴戟天中水晶兰苷、去乙酰基车叶草苷酸、车叶草苷酸和车叶草苷4个成分在各自质量浓度范围内线性关系良好,精密度、重复性、稳定性和加样回收率均符合含量测定要求。巴戟天环烯醚萜苷类成分广西产质量分数为（17.74±0.49）～（60.67±0.28）mg/g,广东产质量分数为（28.46±0.68）～（58.29±0.38）mg/g,福建产质量分数为（20.75±0.42）～（43.94±1.10）mg/g。总量以7年生广西梧州产最高,为（60.67±0.28）mg/g,其次是3年生广东高要产,为（58.29±0.38）mg/g。结论 通过23批不同产地及生长年限的巴戟天药材环烯醚萜苷类成分含量对比,发现7年生广西梧州产巴戟天环烯醚萜苷类成分含量最高,3年生广东高要次之,为巴戟天药材合理的采收时间和质量控制提供参考依据。     

Tumoral cytotoxic and antioxidative phenylpropanoid glycosides in Smilax riparia A. DC

Ethnopharmacological relevance

Smilax riparia A. DC., known as “Niu-Wei-Cai” in China, is distributed through the south and middle of China. The roots and rhizomes of Smilax riparia have been used not only as traditional Chinese medicines (TCMs) for the treatment of bronchitis, lumbago of renal asthenia, traumatic injury, asthenia edema, and cancer but also as edible wild herbs in some areas of China.

Aim of the study

To identify the phytochemicals in the roots and rhizomes of Smilax riparia and to investigate their antioxidant activities and cytotoxicities toward several tumor cell lines.

Materials and methods

Four fractions and five phenylpropanoid glycosides were obtained from roots and rhizomes of Smilax riparia under bioassay-guided screenings. The structures of five compounds were elucidated by spectroscopic methods and compared with published data. We evaluated their antioxidant activities and their cytotoxicities on five cancer cell lines: human promyelocytic leukemia (HL-60), human hepatocellular carcinoma (SMMC-7721), human lung cancer (A-549), human breast cancer (MCF-7), and human colon cancer (SW480).

Results

Of the five glycosides, one new compound (3, smilaside P) was isolated from an EtOAc fraction. Compound 1 was cytotoxic toward HL-60, SMMC-7721, A-549, MCF-7, and SW480 (IC₅₀ 2.70, 3.80, 11.91, 3.79, and 3.93 μM, respectively). Moreover, compounds 1–3 showed moderate scavenging activities against the 2, 2-diphenyl-1-picrylhydrazyl (DPPH) radical (IC₅₀ 339.58, 330.66, 314.49 μM, respectively).

Conclusions

Five phenylpropanoid glycosides were reported for the first time from this TCM. Each was studied, as observed here for the first time, in the cytotoxic experiments toward HL-60, SMMC-7721, and SW480 cell lines. Compound 1, bearing three feruloyl groups and three acetyl groups, had the greatest cytotoxicity toward the five tumor cell lines. Compounds 1–3 showed moderate antioxidant activities. All results reflect that compounds 1–3 are cytotoxic for a wide variety of cancer cell lines of differing tissue origins and that the cytotoxicities of these compounds may be related to their antioxidant activities.     

Acylated iridoid and phenylethanoid glycosides from the aerial parts of Scrophularia nodosa.

From the aerial parts of Scrophularia nodosa L. (Scrophulariaceae), eighteen acylated iridoid glycosides and nine phenylethanoid glycosides were isolated. The structures of eight new iridoid and two new phenylethanoid glycosides were elucidated on the basis of chemical and spectroscopic evidences.     

素馨花总环烯醚萜苷质量标准的研究

目的 建立素馨花的总环烯醚萜苷提取物的质量标准.方法 采用颜色反应和TLC(薄层色谱)对素馨花提取物中总环烯醚萜苷进行定性鉴别;用紫外分光光度法测定了有效部位总环烯醚萜苷的含量;用高效液相色谱法测定了橄榄苦苷的含量.结果 在TLC图谱中可检出橄榄苦苷的特征斑点;建立了UV法测定总环烯醚萜苷含量的分析方法,总环烯醚萜苷的标准曲线为:A =0.0289C-0.0049(R2=0.9998),3.96 ～19.80 μg/ml的范围内线性关系良好,平均回收率为102.64％,RSD=0.65％(n=6);建立了HPLC法测定橄榄苦苷含量的分析方法,橄榄苦苷的标准曲线为:Y=133 959X+23.44 (R2 =0.999 8),0.02 ～0.10 mg·ml-1范围内线性关系良好,平均回收率为100.12％,RSD=0.883％(n=6).结论 该方法稳定、准确、精密度高、专属性强、操作简便可行,适用于提取工艺和纯化工艺优化过程中的含量测定.     

Effects of the ethanolic extract of Spirulina maxima on endothelium dependent vasomotor responses of rat aortic rings

Dietary Spirulina decreases, endothelium-dependently, the responses to vasoconstrictor agonists and increases the endothelium-dependent, agonist-induced, vasodilator responses of rat aorta rings. The aim of this study was to analyze, in vitro, the effects of a raw ethanolic extract of Spirulina maxima on the vasomotor responses of rat aortic rings to phenylephrine and to carbachol. On rings with endothelium, the extract produced the following effects: (a) a concentration-dependent (60-1000 microg/ml) decrease of the contractile response to phenylephrine; (b) a rightward shift and a decrease in maximal developed tension, of the concentration--response curve to phenylephrine; (c) a concentration dependent relaxation of phenylephrine-precontracted rings. These effects were blocked by L-NAME, and not modified by indomethacin. The extract had no effect on the concentration-response curve to carbachol of rings with endothelium. On endothelium-denuded rings the extract caused a significant rightward shift of the concentration response curve to phenylephrine without any effect on maximal tension development. In the presence of the extract, indomethacin induced a marked decrease in the maximal phenylephrine-induced tension of endothelium-denuded rings. These results suggest that the extract increases the basal synthesis/release of NO by the endothelium and, also, the synthesis/release of a cyclooxygenase-dependent vasoconstricting prostanoid by vascular smooth muscle cells.     

UPLC/Q-TOF-MS analysis of iridoid glycosides and metabolites in rat plasma after oral administration of Paederia scandens extracts

A rapid and validated UPLC-MS method was developed for investigating the absorbed components of Paederia scandens(Lour.) Merrill(P.scandens) in rat plasma.The bioactive constituents in plasma samples from rats administrated orally with P.scandens extract were analyzed by Ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF-MS).Four prototype compounds were identified in rat serum as potential bioactive components of P.scandens by comparing their retention times and mass spectrometry data or by mass spectrometry analysis and retrieving the reference literatures.Glucuronidation after deglycosylation was the major metabolic pathway for the iridoid glycosides in P.scandens.These results showed that the methods had high sensitivity and resolution and were suitable for identifying the bioactive constituents in plasma after oral administration of P.scandens.providing helpful chemical information for further pharmacological and mechanistic researched on the P.scandens.     

栀子总环烯醚萜苷对脑出血大鼠内皮屏障的影响

目的:探讨栀子总环烯醚萜苷对脑出血大鼠内皮屏障的影响。方法:SD大鼠随机分为假手术组、模型组和栀子总环烯醚萜苷(30、15和5mg/kg)组,采用自体血注入诱发大鼠脑出血模型,术后24、48、72和120h时间点各组各取10只大鼠脑组织标本,采用免疫组织化学染色检测脑出血周围血肿组织中内皮屏障抗原(endothelial barrier antigen,EBA)和血管内皮整合素αVβ3(αVβ3-Integrin)表达,ELISA法测定促血管生成素-1(angiopoietin-1,Ang-1)和血栓调解蛋白(Thrombomodulin,TM)含量变化。结果:与假手术组比较,术后24～120h模型大鼠脑血肿周围组织中EBA免疫消失区面积增多,整合素αVβ3平均光密度值增加,Ang-1和TM浓度升高。与模型组比较,30mg/kg栀子总环烯醚萜苷组在术后24～120h各时间点EBA免疫消失区面积和整合素αVβ3表达降低,Ang-1和TM浓度降低。15mg/kg栀子总环烯醚萜苷组仅在术后24～72h有效。结论:栀子总环烯醚萜苷可保护或修复内皮屏障,减少EBA消失区面积和整合素αVβ3表达,降低Ang-1和TM水平。     

藏药1号水提液对大鼠离体胸主动脉条收缩作用的影响

目的：通过观察藏药1号水提液对大鼠离体胸动脉条收缩作用的影响研究其降压机制。方法：观察藏药1号水提液(6mg／mL)和维拉帕米(Ver0.013mg／mL)对高K^ 液引起的主动脉条收缩的时效影响，对KCl，NE及CaCl2引起的大鼠主动脉条收缩的量效曲线的影响，以及对NE引起的依赖于细胞内钙及细胞外钙的收缩的影响。结果：藏药1号水提液抑制高K^ 液引起的主动脉收缩；且可使KCl、NE及CaCl2引起的大鼠主动脉条收缩的量效曲线非平行右移，最大效应降低，呈非竞争性拮抗作用：与维拉帕米相似，对NE引起的依赖于细胞内钙及细胞外钙的收缩均有抑制作用。结论：提示藏药1号的降压机制与钙离子通道拮抗剂一致。     

栀子总环烯醚萜苷对脑出血大鼠血肿周围NF-кB表达的影响

目的:研究栀子总环烯醚萜苷对大鼠脑出血后核转录因子кB(NF-кB)表达影响.方法:将SD大鼠随机分为假手术组、脑出血模型组、尼膜同组、栀子总环烯醚萜苷(20、10mg、5mg/kg)治疗组,采用自体血注入诱发大鼠脑出血模型,每组分别在术后12、24、48和96h 4个时间点取脑组织标本,免疫组化检测NF-кB蛋白表达,同时测量脑组织含水量.结果:20～10 mg/kg栀子环烯醚萜苷治疗组脑出血后NF-кB P65表达水平及脑含水量明显低于脑出血组.结论:栀子环烯醚萜苷可以抑制脑出血后NF-кB活化,减少炎症因子释放,从而减轻脑水肿.     

中药中3类环烯醚萜苷抗炎活性研究进展

环烯醚萜苷在传统中药中分布广泛,具有抗炎等多种生物活性,也是中药中重要的一类质控成分。中药中常见的环烯醚萜苷分为九碳骨架环烯醚萜苷、十碳骨架环烯醚萜苷及裂环环烯醚萜苷3类。以3类环烯醚萜苷中15个受到广泛关注的代表性成分,包括京尼平苷、梓醇、龙胆苦苷等为对象,概括了近5年其抗炎功效及可能的相关机制,以期总结不同类型环烯醚萜苷成分的抗炎作用特点。通过对比结构和作用特点分析,发现这些环烯醚萜苷的抗炎作用多与核转录因子-κB(NF-κB)通路和丝裂原活化蛋白激酶(MAPK)通路相关,对肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)、IL-1β这几个炎症因子均有明显的抑制效果。另外,少数具有降低诱导型一氧化氮合酶(i NOS)和环氧合酶-2(COX-2)的表达,通过NLRP3、Nrf2/HO-1、PI3K等通路发挥作用的特点。从构效关系来看,环烯醚萜苷中环戊烷上的双键、C-11取代基、开环后的成键方式均对其抗炎活性存在重要影响。     

Anti-inflammatory effects of iridoid glycosides fraction of Folium syringae leaves on TNBS-induced colitis in rats

Aim of the study

To investigate the effects and the protective mechanism of iridoid glycosides (IG) enriched from Folium syringae leaves on ulcerative colitis (UC) model induced by 2,4,6-trinitrobenzenesulfonic acid (TNBS) in rats.

Materials and methods

UC in rats was induced by colonic administration with TNBS. IG (80, 160 and 240 mg/kg) was administered for 2 week to experimental colitis rats. The inflammatory degree was assessed by macroscopic score, histology and myeloperoxidase (MPO) activity. Nitric oxide (NO) and malondialdehyde (MDA) levels were measured with biochemical methods. The protein expressions of nuclear factor-kappaBp65 (NF-κBp65) and mRNA expressions of pro-inflammatory cytokines, such as tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and NF-κBp65, were determined by immunohistochemistry and real-time quantitative PCR, respectively.

Results

IG significantly ameliorated macroscopic damage and histological changes, reduced the activity of MPO, depressed MDA and NO levels and effectively inhibited the protein and mRNA expressions of NF-κBp65, TNF-α and IL-6 in the colon tissues of experimental colitis in a dose-dependent manner. Moreover, the effects of IG (160 mg/kg and 240 mg/kg) were superior to salicylazosulfapyridine (150 mg/kg).

Conclusion

We demonstrated for the first time that IG possessed marked protective effects on experimental colitis through its antioxidation and inhibiting inflammatory mediators by down-regulation of the expressions of NF-κBp65.     

大孔树脂富集纯化藏药独一味总环烯醚萜苷

目的：进行藏药独一味止血活性成分总环烯醚萜苷的富集纯化。方法：以一阶导数分光光度法检测,以总环烯醚萜苷含量为跟踪指标,比较10种不同厂家、不同型号的大孔树脂对独一味水提取物中环烯醚萜苷类成分的吸附与解吸附条件,优选最佳的纯化工艺。结果：富集后总环烯醚萜苷的含量达到67.53％。结论：XDA-1型大孔吸附树脂对独一味水提取物中环烯醚萜苷类成分有较好的吸附能力,50％乙醇溶液解吸附快速有效。树脂重复利用度高。     

Pharmacological study of phenylpropanoid glycosides: Platelet aggregation and blood pressure studies in rabbits and rats

The effects of phenylpropanoid glycosides (PhGs) isolated from Calceolaria and Orobanche on platelet aggregation and blood pressure have been investigated. The PhGs tested were able to interfere with platelet aggregation, producing pro- or antiaggregant effects. On the other hand, none of the PhGs tested had a significant effect on Mood pressure. The results suggest that the effects of PhGs on rabbit platelets may be related to a differential modulation of cAMP-phosphodiesterase.     

Analysis of the mechanisms underlying the vasorelaxant action of coptisine in rat aortic rings

The aim of the present study was to evaluate the vasorelaxant effects of coptisine and its possible mechanisms in isolated rat aortic rings. Coptisine was evaluated on isolated rat aortic rings precontracted with norepinephrine (NE) and KCl. The mechanisms were evaluated in the presence or absence of specific pharmacological inhibitors. Coptisine (1 ~ 200 μM) relaxed NE (1 μM) or KCl (60 mM) induced sustained contraction with pEC(50) values of 4.49 ± 0.48 and 4.85 ± 0.57 in a concentration dependent manner. Pretreatment with coptisine (10, 50 or 100 μM) also inhibited concentration-response curves to NE and KCl. The vasorelaxant effect of coptisine was attenuated significantly by endothelium removal, and incubation with Nω-nitro-L-arginine methyl ester (L-NAME, 100 μM), methylene blue (10 μM) and indomethacin (5 μM) partially reduced the vasorelaxant effect of coptisine. In endothelium-denuded rings, the vasorelaxant effect of coptisine was reduced significantly by 4-aminopyridine (4-AP, 100 μM), but not glibenclamide (10 μM) ortetraethylammonium (TEA, 5 mM). Coptisine also reduced NE-induced transient contraction in Ca(2+)-free solution, and inhibited contraction induced by increasing external calcium in Ca(2+)-free medium plus 60 mM KCl. It was concluded that coptisine induced both endothelium-dependent and -independent relaxation in rat aortic rings. The NO-cGMP mediated pathway may be involved in the endothelium-dependent relaxation and in the activation of voltage-dependent K(+) channels, contributing in part to the endothelium-independent relaxation bycoptisine. Coptisine also blocks extracellular Ca(2+) influx by interacting with both voltage- and receptor-operated Ca(2+) channels.     

栀子总环烯醚萜苷对大鼠脑出血AQP4和MMP-2/9表达的影响

目的:探讨栀子总环烯醚萜苷对大鼠脑出血AQP4和MMP-2/9表达的影响。方法:SD大鼠随机分为假手术组、模型组和栀子总环烯醚萜苷(30mg/kg)治疗组,采用自体血注入诱发大鼠脑出血模型,术后24、48、72和120h时间点各组各取5只大鼠脑组织标本,采用干湿重法、免疫组织化学染色、逆转录聚合酶链反应技术分别测定脑含水量、AQP4蛋白和AQP4mRNA的表达、MMP-2和MMP-9表达。结果:与模型组比较,治疗组大鼠在各时间点脑含水量减少(P0.01),AQP4蛋白和AQP4mRNA表达降低(P0.05),术后72～120h时间点MMP-2和MMP-9表达水平低(P0.05～P0.01)。结论:栀子总环烯醚萜苷可能通过抑制AQP4、MMP-2和MMP-9的表达减轻脑出血后脑水肿的形成。