Trillin, a steroidal saponin isolated from the rhizomes of Dioscorea nipponica, exerts protective effects against hyperlipidemia and oxidative stress

Ethnopharmacological evidence

Numerous efforts have been conducted in searching for effective agents against cardiovascular diseases, in particular from herbal medicines. The rhizome of Dioscorea nipponica (Dioscoreae Nipponicae Rhizoma) is a traditional Chinese herb being prescribed to improve the blood circulation. Here, we identified a steroidal saponin trillin from Dioscorea nipponica, which showed robust anti-hyperlipidemic effects.

Materials and methods

Rats were induced for hyperlipidemia and subjected to the drug treatment. The anti-hyperlipidemic effects of trillin were evaluated by different biochemical assays.

Results

In hyperlipidemic rat model, fed with high-fat diet, the blood levels of cholesterol, triglyceride, low density lipoprotein (LDL) and high density lipoprotein (HDL) were increased. The intra-peritoneal administration of trillin into those rats significantly improved the bleeding and blood coagulation time, and in parallel the treatment restored the levels of cholesterol, glyceride, LDL and HDL back to the normal condition. In addition, the administration of trillin in rats exerted beneficial effects in improving the levels of lipid peroxidation and superoxide dismutase activity.

Conclusion

This was the first time to reveal the anti-hyperlipidemic and anti-oxidative effects of trillin. These results would be important in developing food supplements for health improvements and therapeutic drugs against hyperlipidemia and cardiovascular diseases in future.     

德国鸢尾化学成分研究

通过硅胶,Sephadex LH-20葡聚糖凝胶,ODS反相柱色谱等分离技术对鸢尾科Iridaceae植物德国鸢尾Iris germanica根茎的化学成分进行分离纯化,采用理化性质及波谱分析方法对分离得到的化合物进行结构鉴定。从该植物根茎中分离得到21个化合物,分别鉴定为商陆素（1）,5,3,3’-三羟基-7,4’-二甲氧基黄烷酮（2）,5,7,4’-三羟基双氢黄酮（3）,cirsiliol-4’-glucoside（4）,3β,4’-dihydroxy-7,3’-dimethoxy flavornone-5-O-β-D-glucopyranoside（5）,染料木素（6）,irilin D（7）,muningin（8）,5,7, 4’-三羟基-6,3’,5’-三甲氧基异黄酮（9）,鸢尾苷元（10）,野鸢尾苷元（11）,鸢尾苷（12）,野鸢尾苷（13）,芒果苷（14）,irisxanthone（15）,焦谷氨酸（16）,2,4’, 6-trihydroxy-4-methoxybenzophenone-2-O-β-D-glucoside（17）,茶叶花宁（18）,草夹竹桃苷（19）,β-谷甾醇（20）,胡萝卜苷（21）。其中化合物 1~9,16,17 为首次从德国鸢尾中分离得到,化合物 8,9 为首次从鸢尾属植物中分离得到,化合物 1,4,17 为首次从鸢尾科植物中获得。     

小槐花大极性部位的化学成分研究

该实验研究了传统中药小槐花大极性部位的化学成分。综合运用硅胶柱色谱,聚酰胺凝胶柱色谱,ODS中低压柱色谱,Sephadex LH-20柱色谱,薄层色谱,重结晶,分析型和制备型HPLC等方法进行分离,利用核磁共振波谱,质谱及化合物的理化性质对化合物的结构进行了鉴定。从小槐花大极性部位中分离得到13个单体化合物,它们的结构经鉴定分别为香草醛（1）,黑麦草内酯（2）,吲哚-3-甲醛（3）,水杨酸（4）,当药黄素（5）,saccharumoside C（6）,isosinensin（7）,7-O-α-L-吡喃鼠李糖基-山柰酚-3-O-β-D-吡喃葡萄糖苷（8）,异牡荆素（9）,牡荆素（10）,nothofagin（11）,resveratroloside（12）,2"-α-rhamnopyranosyl-7-O-methylvitexin（13）。除化合物5外,其余12个化合物均为小槐花中首次分离得到的化合物,化合物2,3,6~8,11~13等8个化合物为山蚂蝗属植物中首次分离得到的化合物。     

穿龙薯蓣淀粉的理化性质研究

目的 为了充分了解穿龙薯蓣,通过多种分析方法对穿龙薯蓣淀粉物理化学性质进行研究.方法 采用扫描电子显微镜(SEM)、粒度分析仪、X-射线衍射、差示扫描量热仪(DSC)和布拉班德粘度仪.结果 通过与木薯、马铃薯淀粉的物理化学性质的比较,穿龙薯蓣淀粉显示了相对较小、椭圆形,大小不均匀的颗粒.晶体类型显示为C型.含直链淀粉为26.3%.凝胶化温度最高,其焓值介于马铃薯与木薯.根据布拉班德图谱得知穿龙薯蓣淀粉具有较低的黏度峰值、较高的回生值和较低的衰退值.结论 穿龙薯蓣淀粉与其他相关植物的块茎类淀粉来源不同,物理化学性质也存在明显不同.     

Anti-oxidant and inflammatory mediator's growth inhibitory effects of compounds isolated from Phyllanthus urinaria

Phyllanthus urinaria Linnea (Euphorbiaceae), is a traditional anti-hepatitis herb used in Taiwan. In continuation of our search for potent natural anti-inflammatory agents, from the ethanolic extract of this plant, nine compounds including phyllanthin (1), phyltetralin (2), trimethyl-3,4-dehydrochebulate (3), methylgallate (4), and rhamnocitrin (5), methyl brevifolincarboxylate (6), beta-sitosterol-3-O-beta-d-glucopyranoside (7), quercitrin (8), and rutin (9) were isolated. The structures of compounds 3 and 6 were established based on NMR and mass spectral studies. The isolates 1-9 were investigated for their antioxidant, and anti-inflammatory activities in vitro. In the antioxidant assay, the isolates 3, 4 and 6 exhibited significant DPPH radical scavenging activity with an IC(50) value of 9.4, 9.8 and 8.9 microM, respectively. On the other hand, in the inflammatory mediators growth inhibitory assay from LPS/interferon (IFN)-gamma-activated peritoneal macrophages, all the isolates except 7, significantly and dose-dependently inhibited the enhanced production of NO radicals, and such modulation was closely associated with the inhibition of tumor necrosis factor (TNF)-alpha and interleukin (IL)-6. In addition, 30 microM of isolates 3 and 6, and 50 microM of 4, significantly arrest the mitogen-stimulated spleen cells in G0/G1 stage. This is the first report on Phyllanthus urinaria isolates for their growth inhibitory activities against inflammatory mediators, in addition to spleen cell cycle arrest in G0/G1 stage. Therefore, these isolates from Phyllanthus urinaria may be useful for the treatment of cell-mediated immune diseases.     

新疆紫草羟基萘醌类成分对炎症细胞因子的抑制作用

目的研究新疆紫草羟基萘醌化合物对炎症细胞因子的抑制作用。方法采用酶联免疫法测定新疆紫草羟基萘醌化合物对脂多糖（LPS）诱导的小鼠腹腔巨噬细胞RAW264．7释放肿瘤坏死因子-α（TNF-α）和白细胞介素-1D（IL-1D）等炎症细胞因子的抑制作用。采用Griess法测定分离纯化的6种不同羟基萘醌化合物（紫草素、去氧紫草素、乙酰紫草素、β’β-二甲基丙烯酰紫草素、α-甲基丁酰紫草素、异丁酰紫草素）对LPS诱导的小鼠腹腔巨噬细胞RAW264．7释放炎症细胞因子一氧化氮（N0）的抑制作用。结果新疆紫草羟基萘醌化合物对LPS诱导的小鼠腹腔巨噬细胞RAW264．7释放TNF—α和IL-1D具有抑制作用;6种紫草羟基萘醌化合物对LPS诱导的小鼠腹腔巨噬细胞RAW264．7释放NO具有抑制作用。结论新疆紫草羟基萘醌成分对炎症细胞因子均有不同程度的抑制作用,为进一步研发以炎症因子TNF-α抑制剂为靶向的类风湿关节炎治疗药物提供了一定的理论依据。     

Immunomodulatory and antitumor activity of triterpenoid fractions from the rhizomes of Astilbe chinensis

Aim of the study

The objectives of this study were to evaluate the in vivo antitumor potential of the triterpenoid fraction from the rhizomes of Astilbe chinensis (Saxifragaceae) (Saxifragaceae) (ATF) and to elucidate its immunological mechanisms by determining its effects on the growth of mouse transplanted tumors and the immune response in naïve and tumor-bearing mice.

Materials and methods

The mice inoculated with mouse tumor cell lines were treated per os with ATF at the doses of 20, 40, 60 mg/kg for 10 days. The effects of ATF on the growth of transplantable tumor, splenocyte proliferation, the activity of natural killer (NK) cells, and production of interleukin-2 (IL-2) from splenocytes in tumor-bearing mice were measured. Meanwhile, the effects of ATF on 2,4-dinitrofluorobenzene (DNFB)-induced delayed type hypersensitivity (DTH) reaction and the sheep red blood cell (SRBC)-induced antibody response in naïve mice were also studied.

Results

ATF could not only significantly inhibit the growth of mice transplantable tumor, but also remarkably increase splenocytes proliferation, NK cells activity, and the level of IL-2 secreted by splenocytes in tumor-bearing mice, promote the DTH reaction and enhance anti-SRBC antibody level in naïve mice, which indicated that the ATF could improve both specific and non-specific cellular and humoral immune response.

Conclusions

The antitumor activity of ATF might be achieved by improving immune response, and ATF could act as antitumor agent with immunomodulatory activity.     

石菖蒲的化学成分研究

对石菖蒲Acorus tatarinowii根茎的化学成分进行研究.应用硅胶,ODS,Sephadex LH-20,HPLC等各种色谱技术进行分离和纯化,采用NMR等谱学方法鉴定结构.从石菖蒲根茎的95％乙醇提取物的乙酸乙酯萃取物中分离得到15个化合物,分别鉴定为tatanone A(1),水菖蒲酮(2),菖蒲螺酮烯(3),2-acetyloxyacoronene(4),菖蒲螺烯酮(5),α-细辛醚(6),β-细辛醚(7),顺式-甲基异丁香酚(8),甲基丁香酚(9),细辛醛(10),细辛酮(11),γ-细辛醚(12),5-羟甲基糠醛(13),galgravin (14)和桉脂素(15).其中化合物1为新化合物,化合物3～5为首次从该属植物中分得.     

三七炮制品化学成分研究

为了研究三七炮制品的化学成分,采用大孔吸附树脂柱色谱、硅胶柱色谱、RP-C₁₈柱色谱、制备HPLC等分离纯化方法,从三七炮制品的70%乙醇提取物中分离得到23个单体化合物。应用理化测定和波谱(MS,NMR)分析等方法鉴定各化合物的结构,其分别鉴定为6'-O-乙酰人参皂苷Rh₁(1),人参皂苷Rk₃(2),人参皂苷Rh₄ (3),20S-人参皂苷Rg₃(4),人参皂苷Rk₁(5),20R-人参皂苷Rg₃(6),人参皂苷Rg₅ (7),人参皂苷F₂ (8),20S-人参皂苷Rh₁(9),20R-人参皂苷Rh₁(10),绞股蓝皂苷ⅩⅦ(11),三七皂苷Fa(12),人参皂苷Ra₃(13),人参皂苷Rg₁ (14),人参皂苷Re(15),三七皂苷R₂(16),20R-人参皂苷Rg₂(17),三七皂苷R₁ (18),人参皂苷Rd(19),人参皂苷Rb₁ (20),三七皂苷D(21),三七皂苷R₄(22)和人参皂苷Rb₂(23)。化合物 1 为首次从三七中分离得到,化合物 4, 6, 8和11 为首次从三七炮制品中分离得到。根据三七炮制前后样品的指纹图谱分析比对,推测了在炮制过程中三七中人参三醇类和人参二醇类化合物可能的变化途径。结果显示,三七中的三萜皂苷主要发生了糖基水解反应和脱水反应。     

红茴香根茎的化学成分研究

目的:研究八角科Illiciaceae植物红茴香Illicium henryi根茎的化学成分。方法:应用硅胶,Sephadex LH-20,Rp-8,Rp-18等各种色谱技术进行分离纯化,根据化合物的理化常数和波谱数据鉴定其结构。结果:从红茴香根茎中分离得到12个化合物,分别鉴定为蛇菰脂醛素(1),aviculin(2),rubriflosides A(3),1,2-bis(4-hydroxy-3-methoxyphenyl)-1,3-propanediol(4),jasopyran(5),山柰酚(6),槲皮素(7),(2R,3R)-3,5,7,3′,5′-五羟基黄烷(8),3,4,5-trimethoxyphenyl-1-O-β-D-gluco-pyranoside(9),3,4-dimethoxyphenyl-1-O-β-D-glucopyranoside(10),松柏醛(11),芥子醛(12)。结论:化合物1~12均为首次从该植物中分离得到。     

Bioactivity-guided isolation of polyacetylenes with inhibitory activity against NO production in LPS-activated RAW264.7 macrophages from the rhizomes of Atractylodes macrocephala

Ethnopharmacological relevance

The rhizome of Atractylodes macrocephala (Compositae) is one of the most well-known traditional Chinese medicine in China, Japan and Korea, which has a long history of use for the treatment of splenic asthenia, edema, anorexia, and excessive perspiration, etc. As active compounds of anti-inflammatory activity of this medicinal plant have not been fully elucidated, the aim of this study was to isolate and identify the active constituents inhibiting nitric oxide (NO) production from the rhizomes of A. macrocephala.

Materials and methods

Inhibitory activity against NO production in lipopolysaccharide-activated RAW264.7 macrophages was evaluated by Griess reaction. Fifteen polyacetylenes were isolated from the active ethyl acetate extract using activity-guided screening. The structures of all compounds were elucidated by spectroscopic methods and comparison with published data. The compounds were further tested for their inhibitory activity against NO production.

Results

Seven new polyacetylenes, named atractylodemaynes A–G (1–7), along with eight known ones (8–15) were isolated. Compound 14 was isolated for the first time from the rhizomes of A. macrocephala. The study showed that the tested compounds exhibited inhibitory activity against NO production in a dose-dependent manner. Among them, compounds 10, 11 and 12 had relatively stronger inhibitory effect with IC₅₀ values of 28, 23 and 19 μM, respectively.

Conclusion

The results demonstrated that the polyacetylenes might greatly contribute to the anti-inflammatory activity of the rhizomes of A. macrocephala.     

裂叶羌活中1个新的天然产物O-甲基羌活醇

目的:研究裂叶羌活Notopterygium,incisum干燥根和根茎甲醇提取物的乙酸乙酯溶性部位化学成分.方法:采用多种色谱方法对其化学成分进行分离和纯化,核磁共振波谱法和质谱法鉴定其结构.结果:从裂叶羌活甲醇提取物的乙酸乙酯溶性部位分离得到5个化合物,分别鉴定为镰叶芹二醇(1),O-甲基二氢镰叶芹酮醇(2),川白芷素(3),东莨菪素(4),O-甲基羌活醇(5).结论:化合物5为新的天然产物;化合物2～4为首次从裂叶羌活根和根茎中分离得到.     

Cytochrome P450 inhibitory potential of Triphala--a Rasayana from Ayurveda

Ethnopharmacological relevance

‘Triphala’ is one of the age-old, most commonly used polyherbal preparation from Ayurveda as Rasayana drug.

Aim of the study

This study was aimed at evaluating the effect of ‘Triphala’ on drug modulating enzymes to assess its safety through its potential to interact with co-administered drugs.

Materials and methods

The cytochrome P450 inhibitory effect of ‘triphala’ formulation was investigated on rat liver microsomes using CYP450-CO complex assay and on individual isoform such as CYP3A4 and 2D6 using fluorescence screening. RP-HPLC method was developed to standardize ‘triphala’ and its individual components using gallic acid as analytical marker compound.

Results

RP-HPLC analysis demonstrated the presence of gallic acid (4.30 ± 2.09 mg/g) in the formulation. The formulation showed 23% inhibition of the rat liver microsomes through CYP450-CO complex assay which is comparatively less when compared with the individual components. Further, the effect of standardized formulation dissolved in ethanol showed CYP3A4 and CYP2D6 inhibitory activity at the IC₅₀ values of 119.65 ± 1.91 μg/ml and 105.03 ± 0.98 μg/ml respectively. Gallic acid was also found to inhibit both the isoforms at the IC₅₀ values of 87.24 ± 1.11 μg/ml and 92.03 ± 0.38 μg/ml respectively.

Conclusions

Various concentrations of the formulation and its individual components showed significantly less inhibitory activity (p < 0.001) on individual isoforms when compared with the positive control. Assessment on the in vitro effect of ‘triphala’ on drug modulating enzymes has important implications for predicting the likelihood of herb-drug interactions if these are administered concomitantly.     

盾叶薯蓣鲜根茎中甾体皂苷的分离鉴定

目的 研究盾叶薯蓣Dioscorea zingiberensis鲜根茎中的甾体皂苷成分,寻找新的生物活性物质.方法 运用正相和反相C18硅胶柱色谱对其甾体皂苷成分进行分离,用IR、MS、NMR等光谱方法进行结构鉴定.结果 从乙醇提取物中分离鉴定了3个甾体化合物,分别为纤细皂苷(gracillin,Ⅰ)、盾叶皂苷A(zingiberenin A,Ⅱ)和26-O-βD-吡喃葡萄糖基-3β,26-二醇-25(R)-呋甾烷-5,20(22)-双烯-3-O-{α-L-吡喃鼠李糖基-(1→4)-[β-D-吡喃葡萄糖基-(1→3)-β-D-吡喃葡萄糖基-(1→2)])-β-D-吡喃葡萄糖苷(26-O-β-D-glucopyranosyl-3β,26-dioi-25(R)-furosta-5,20(22)-dien-3-O-{α-L-rhamnopyranosyl-(1→4)-[β-D-glucopyranosyl-(1→3)-β-D-glucopyranosyl-(1→2)]}-β-D-glucopyranoside,Ⅱ).结论 化合物Ⅰ为一新化合物.命名为盾叶皂苷G(zingiberenin G).     

山药微型块茎诱导形成的影响因子研究

目的 探讨培养方式、碳源、氮源对山药微型块茎诱导形成的影响,以期找到适宜的微型块茎诱导形成的培养方式和培养基.方法 以B号山药试管苗为材料,运用一步培养法和两步培养法筛选最佳培养方式,在此基础上于培养基中添加不同氮源和碳源进行诱导培养,定期对微型块茎的诱导情况进行统计.结果 两步培养法、60g/L的碳源(白砂糖或蔗糖)、[NO3-]/[NH4 ]为3:1、总氮量为30 mmol/L时有利于山药微型块茎的诱导形成.结论 首次建立了两步培养法诱导山药微型块茎形成的培养模式,筛选出了适宜的碳源和氮源,为微型块茎的离体诱导形成及工厂化生产奠定了技术基础.     

黑刺菝葜根茎中苯丙素和二苯乙烯类化学成分的研究

运用硅胶,Sephadex LH-20等各种色谱技术对黑刺菝葜Smilax scobinicaulis根茎的化学成分进行研究.通过波谱数据的分析鉴定化合物的结构分别为白藜芦醇3-O-β-D-葡萄糖基-(1→3)-β-D-葡萄糖苷(1),白藜芦醇(2),δ-葡萄素(3),咖啡酸乙酯(4),1-O-咖啡酰甘油酯(5),1-O-对香豆酰基甘油酯(6),1-O-阿魏酰甘油酯(7),大海米酰胺(8),桑辛素M(9).其中化合物1为新化合物,化合物3～5,8,9为首次从该植物中分离得到.     

姜的化学成分研究

目的:对姜科姜属植物姜Zingiber officinale根状茎的化学成分进行分离和鉴定.方法:采用硅胶、Sephadex LH20等柱色谱方法对化学成分进行分离和纯化,根据其理化性质及波谱数据鉴定结构.结果:分离鉴定了9个化合物,分别为β-谷甾醇棕榈酸酯(β-sitosterol palmitate,1),异香草醛(isovanillin,2),棕榈酸乙二醇单酯,(glycol monopalmitate,3),二十六烷酸单甘油酯(hexacosanoic acid 2,3-dihydroxypropyl ester,4),马来酸酰亚胺-5-肟(maleimide-5-oxime,5),对羟基苯甲醛(Phydroxybenzaldehyde,6),腺嘌呤(adenine,7),6-姜酚(6-gingerol,8),6-姜烯酚(6-shogaol,9),和1个组合物1-(ω-ferulyloxyceratyl)glycerols(10a～10f),结论:化合物1～7,10a～10e是首次从该植物中分离得到,1-(30-ferulyloxytriaconatanoyl)glycerol(10f)是新化合物.     

长托菝葜化学成分研究

目的:研究民间习用药材长托菝葜根茎的化学成分。方法:采用反复硅胶柱、MCI、Sephadex LH-20和RP C18柱色谱分离纯化,并运用现代波谱分析方法(MS,IR,NMR)鉴定其化学结构。结果:从长托菝葜中分离得到6个化合物,分别鉴定为二氢山柰酚(dihydrokaempferol,1),山柰酚(kaempferol,2),落新妇苷(astibin,3),黄杞苷(engeletin,4),反式白藜芦醇(res-veratrol,5),β-谷甾醇(β-sitosterol,6)。结论:所有化合物均首次从该植物中分离得到。     

山药多糖的提取分离和结构测定

目的：提取分离并测定山药多糖的结构。方法：依次用水提取，乙醇和十六烷基三甲基溴胺盐沉淀法得到粗多糖；用萄聚糖凝胶柱层析和高效液相层析纯化多糖并测定分子量；甲基化分析和薄层层析分析多糖酸水解产物，结合IR和^13CNMR确定单糖组成、连接方式和端基碳构型。结果：从山药中得到两个均-多糖S1和S2。结论：S1和S2的分子量分别为63000和7400Dal，它们均为[α-D—Glc(1→4)-]n型萄聚糖。