An Acidic Polysaccharide from Tribulus terrestris

An aqueous acidic polysaccharide, named rhamnogalacturonan (designated as TTP-D2) was isolated from Tribulus terrestris L by means of DEAE-cellulose chromatography and gel filtration. The molecular mass of TTP-D2 was estimated to be 26 KDa by gel filtration. TTP-D2 is composed of galacturonic acid, rhamnose, arabinose, galactose, fucose, mannose, xylose and glucose in a ratio of 71.4 : 13.5 : 5.6 : 4.9 : 3.1 : 1.9 : 1.9 : 1.0. The main chain structure of TTP-D2 was elucidated as an acidic hetero-polysaccharide with the connection of a-(1-4) galacturonic acid with a-(1-3) rhamnose by GC analysis of partially hydrolyzed products and the determination of 1H, 13C-NMR spectra.

Modulatory effects of the fruits of Tribulus terrestris L. on the function of atopic dermatitis-related calcium channels,Orai1 and TRPV3

Objective: To examine the effects of Tribulus terrestris L.(T. terrestris) extract on the modulation of calcium channels to evaluate its use in topical agents for treatment of atopic dermatitis.Methods: The 70% methanol extract of T. terrestris was prepared. Human HEK293 T cells with over-expressed calcium release-activated calcium channel protein 1(Orai1),transient receptor potential vanilloid 1, or transient receptor potential vanilloid 3(TRPV3)were treated with T. terrestris extract. Modulation of ion channels was measured using a conventional whole-cell patch-clamp technique.Results: T. terrestris extract(100 mg/m L) significantly inhibited Orai1 activity in Orai1-stromal interaction molecule 1 co-overexpressed HEK293 T cells. In addition, T. terrestris extract significantly increased the TRPV3 activity compared with 2-Aminoethyl diphenylborinate(100 mmol/L), which induces the full activation of TRPV3.Conclusions: Our results suggest that T. terrestris extract may have a therapeutic potential for recovery of abnormal skin barrier pathologies in atopic dermatitis through modulating the activities of calcium ion channels, Orai1 and TRPV3. This is the first study to report the modulatory effect of a medicinal plant on the function of ion channels in skin barrier.

Two New Furostanol Saponins from Tribulus Terrestris L.

Two new furostanol glucosides, named tribufurosides G and H, were isolated from Tribulus terrestris L. The structures of the two new furostanol glucosides were elucidated as 26-O-β-D-glucopyranosyl-（25R）-5α-furost12-one-3β,22α,26-tetraol-3-O-β-D-glucopyranosyl-（ 1→2 ）-β-D-glucopyranosyl-（ 1→4 ）-β-D-galactopyranoside（ 1 ） and 26-O-β-D-glucopyranosyl-（25S）-5 α- 12-one-2 α,3,8,22 α,26-tetraol-3 -O-β-D-glucopyranosyl-（ 1 → 2 ）-β-D-glucopyranosyl-（1→4）-β-D-galactopyranoside（2） by 1 D, 2D NMR techniques, ESI-MS analysis as well as chemical methods.

Steroidal saponins with anti-inflammatory activity from Tribulus terrestris L.

Objective:Tribulus terrestris L.(T.terrestris)is a highly valuable traditional Chinese medicine used to treat stroke,inflammation,pulmonary fibrosis,liver cancer,and urolithiasis.To identify the basic substance responsible for the anti-inflammatory effect of TST(total saponins of Tribulus),its chemical composition was systematically studied,and its effect of inhibiting nitric oxide generation and the expression of related inflammatory factors were determined.Methods:To separate chemical constituents from T.terrestris by column chromatography.Spectroscopic methods,including 1D and 2D nuclear magnetic resonance spectroscopy(NMR)and mass spectrometry(MS)techniques,were used to elucidate the isolated compounds.The anti-inflammatory activities of TST and several compounds were evaluated in vitro.Results:Fifteen steroidal saponins,including 9 furostanol steroidal saponins(1,2,3,4,5,6,7,8,and 15)and 6 isospirostanol steroidal saponins(9,10,11,12,13,and 14),were isolated from T.terrestris.TST significantly decreased the expression of tumor necrosis factor-a(TNF-a)and interleukin-6(IL-6)in RAW 264.7 cells stimulated by lipopolysaccharides.Compounds 13 and 15 evidently reduced TNF-a expression.Compounds 6,10,12,13,and 15 markedly reduced IL-6 expression.Conclusions:Compounds 1 was a novel furostanol steroidal saponin,named 26-O-b-D-glucopyranosyl-(25R)-5afurostan-12-carbonyl-20(22)-en-3b,26-diol-3-O-{b-D-xylopyranosyl-(1→2)-[b-D-xylopyranosyl-(1→3)]-b-D-glucopyranosyl-(1→4)-[a-L-rhamnopyranosyl-(1→2)]-b-D-galactopyranoside}.Compounds 2 was isolated from the family Zygophyllaceae for the first time,and 5 and 6 were isolated from the Tribulus genus.TST and compounds 6,10,12,13,and 15 exerts antiinflammatory activity.

Optimization of Extraction of Saponins from Tribulus terrestris L.

To promote the recovery of saponins from Tribulus terrestris L.,the water dissolution and ethanol precipitation method was adopted to extract saponins from the powder prepared from T.terrestris fruit.The optimal process was to extract with 9 fold of the extracting water at the ethanol concentration of 85 for 3 times,each time 3.5 h.The effects of the factors ranked as the extracting water quantity > treatment time > treatment times > ethanol concentration.Under these optimal conditions,the recovery of saponins was 2.81%.After these crude saponins were purified by D101 macroporous adsorptive resin,the purity was found to be promoted through the UV scanning method,and the recovery of the refined saponins was up to 1.53%.Both of the recovery values were higher than that of the existing process.

RP-HPLC比较蒺藜全草和果实中的主要甾体皂苷元成分

目的:比较蒺藜全草和果实中主要甾体皂苷元成分,探索蒺藜中的确切指标成分。方法:采用反相高效液相色谱法,利用C18柱和示差折光检测器,定量比较了蒺藜(TribulusTerrestrisL. )全草和果实中的主要甾体皂苷元成分。结果:全草与果实中的海可皂苷元含量相近而替告皂苷元含量有显著差异 (P<0. 01)。结论:替告皂苷可能是蒺藜全草的主要指标成分。

心脑舒通胶囊对高脂血症大鼠心肌梗死后心肌细胞凋亡及Bax、Bcl-2蛋白表达的影响

目的观察心脑舒通胶囊(主要起效成分为蒺藜总皂苷)对高脂血症大鼠心肌梗死(MI)后心肌细胞凋亡及Bax、Bcl-2蛋白表达的影响,探讨蒺藜皂苷抗高脂大鼠MI后心室重构的作用机制。方法利用高脂血症心肌梗死复合模型,采用TUNEL和免疫组化SP法,观察蒺藜总皂苷对MI后大鼠心肌细胞凋亡、以及Bax、Bcl-2蛋白表达的影响。结果(1)蒺藜总皂苷可减轻MI后心肌细胞损伤和心室重构的程度;(2)与高脂心梗组相比,蒺藜总皂苷大剂量、辛伐他汀均降低MI后心肌细胞凋亡指数,并能下调Bax蛋白表达(P<0．05),各治疗组之间作用比较,差异无显著性(P>0．05)。结论蒺藜总皂苷可通过调控Bax／Bcl-2蛋白表达比值,而抑制心肌细胞凋亡,可能是其抗MI后心室重构的作用机制之一。

HPLC法同时测定蒺藜茎叶中3个黄酮醇苷的含量

目的：建立HPLC法同时测定蒺藜茎叶中含量较高的3个黄酮醇苷类化合物的含量。方法：色谱柱：Thermo Syncronis C18（250 mm×4.6 mm，5μm）；流动相：水（A）-乙腈（B），梯度洗脱，0-30 min，8%-27% B；检测波长：254 nm；柱温：40℃；流速：1.0 mL＆#183;min-1；进样量：20μL。结果：异鼠李素-3-O-β-D-龙胆双糖基-7-O-β-D-葡萄糖苷、槲皮素-3-O-β-D-龙胆双糖苷、异鼠李素-3-O-β-D-龙胆双糖苷分别在0.0112-0.2800μg（r=0.9996）、0.0648-2.5920μg（r=0.9998）、0.0184-0.4600μg（r=0.9998）范围内线性关系良好；平均加样回收率（n=9）分别为100.15%（RSD=1.32%）、100.02%（RSD=1.14%）、99.77%（RSD=1.16%）。结论：该方法简便、快速、准确、灵敏，为蒺藜或其制剂的质量控制和综合评价提供参考。

蒺藜总皂甙对TNF-α及IL-1_β诱导内皮细胞粘附的抑制作用

目的：观察蒺藜总皂甙（ｇｒｏｓｓｓａｐｏｎｉｎｓｆｒｏｍ ＴｒｉｂｕｌｕｓｔｅｒｒｅｓｔｒｉｓＬ，ＧＳＴＴ） 对肿瘤坏死因子－ α（ＴＮＦ－ α） 、白细胞介素－１β（ＩＬ－１β） 诱导的培养新生牛脑微血管内皮细胞（ＣＭＥＣ） 粘附大鼠血单核细胞（ＭＮＣ） 及中性粒细胞（ＰＭＮ） 的影响。方法：用ＴＮＦ－α和ＩＬ－１β分别诱导传代的ＣＭＥＣ，计粘附ＭＮＣ及ＰＭＮ的细胞数，并计算粘附率。结果：ＣＭＥＣ 经ＴＮＦ－ α处理４ｈ 后，ＣＭＥＣ与ＭＮＣ及ＰＭＮ的粘附率，分别增加到３１．３±０．５％ 和３２．１±０．５％ （对照组分别为１２ ．５±０ ．２％ 和１３ ．８±０ ．４ ％） ，经ＩＬ－１β处理２ｈ 后，则分别为４４．５ ±１．０ ％ 和４４．８ ±０．７ ％ （ 对照组分别为１２ ．９ ±０．６ ％ 和１４．７ ±０．６％ ）。在ＴＮＦ－ α（ＩＬ－ １β） 处理前，用ＧＳＴＴ与ＣＭＥＣ共孵育，预处理２ｈ，则ＧＳＴＴ可剂量依赖性地抑制ＴＮＦ－ α和ＩＬ－ １β的作用。结论：ＧＳＴＴ可抑制ＣＭＥＣ 与ＰＭＮ及ＭＮＣ的粘附。

蒺藜总皂苷对缺氧复氧心肌细胞TNF-α,IL-1β及ICAM-1表达的影响

目的:观察蒺藜总皂苷(gross saponins from Tribulus terrestris L,GSTT)对缺氧复氧心肌细胞炎症分子肿瘤坏死因子-α(TNF-α),白介素1β(IL-1β)及细胞间黏附分子-1(ICAM-1)表达的影响。方法:利用原代培养的SD乳鼠心肌细胞建立心肌缺氧复氧模型,分为正常组、模型组和GSTT高、低浓度(50,10mg·L^(-1))组。用放免法检测TNF-α和IL-1β,细胞ELISA检测ICAM-1表达。结果:与模型组比较,GSTT高、低浓度(50,10mg·L^(-1))组TNF-α和IL-1β含量显著降低(P＜0．01或P＜0．05);GSTT 50 mg·L^(-1)组ICAM-1蛋白表达量显著降低(P＜0．05)。结论:GSTT对缺氧复氧损伤心肌细胞有保护作用,其机制与抑制炎性因子TNF-α和IL-1β释放,降低ICAM-1蛋白表达有关。

HPLC-ELSD测定心脑舒通胶囊中Terrestrinin A的含量

[目的]建立心脑舒通胶囊及其总皂苷中Terrestrinin A含量的测定方法。[方法]采用高效液相色谱-蒸发光散射检测器(HPLC-ELSD)法,色谱柱为Hedra-C_(18)柱(4.6 mm×150 mm,5μm),流动相:乙腈-水,流速:1 mL/min,柱温:40℃,漂移管温度:60℃,气体(N2)压力:30 psi,增益值:100,进样量:20μL。[结果]线性范围:48.5~970.6μg/mL,回归方程:Y=1.693 1X+1.264 4(r^2≥0.999 0),精密度、重复性和稳定性实验的RSD值分别是1.21%、1.62%、1.90%,加样回收率是96.0%~103.2%,RSD值是2.40%。[结论]该法简便可靠,可以应用于心脑舒通胶囊及其总皂苷中Terrestrinin A的含量测定及质量控制标准建立。

HPLC-DAD法测定蒺藜茎叶3种酸水解黄酮苷元的含量

目的:对酸水解后黄酮类三大苷元质量控制进行研究,建立水解充分完全,操作简单易行的质量控制方法。方法:采用HPLC-DAD法,色谱柱:Thermo Syncronis C18(250 mm×4.6 mm,5μm);流动相:为乙腈-0.2%磷酸水溶液,流速:1.0 m L·min-1,柱温:40℃,进样量:20μL,槲皮素、山奈酚及异鼠李素检测波长:370 nm。结果:槲皮素、山奈酚、异鼠李素3个黄酮苷元分别在1.76-44.00,0.81-10.16,0.55-13.75μg/m L范围内呈线性良好;回归方程分别为Y=74906X-64345(R2=0.9999),Y=74324X-40391(R2=0.9993),Y=69020X-30737(R2=0.9996)。结论:通过提取分离工艺中不同部分的样品进行测定发现,经大孔树脂柱(D101)分离后96%的黄酮类化合物主要集中在30%乙醇部分。

HPLC法测定蒺藜中总黄酮的含量

目的建立测定蒺藜中总黄酮含量的高效液相色谱法。方法以槲皮素、山柰素、异鼠李素为对照品,色谱条件:ODS柱(150 mm×4.6 mm,5μm),流动相为甲醇:0.4磷酸(v/v)45∶55,检测波长为360 nm。结果槲皮素、山柰素、异鼠李素回收率试验的RSD分别为1.59%、0.98%、1.42%。结论本方法简便易行,适用于蒺藜总黄酮的含量测定。

蒺藜药材HPLC指纹图谱研究

目的采用高效液相法建立蒺藜药材的指纹图谱,以控制其质量。方法用反相高效液相色谱(RP-HPLC)-ELSD法测定蒺藜的指纹图谱,并根据向量夹角法计算相似度,进行比较分析。结果初步建立了蒺藜药材的指纹图谱。结论HPLC指纹图谱法重现性好,可作为控制蒺藜药材内在质量的标准。

HPLC测定刺蒺藜总皂苷中的薯蓣皂苷元

目的测定刺蒺藜总皂苷水解后薯蓣皂苷元的含量。方法采用ELSD-HPLC法,并考察其方法学。结果薯蓣皂苷元39.68~942μg.ml-1与峰面积的线性关系良好,方法具有较好的精密度、稳定性、重复性和较高的回收率。结论所建方法准确、简便、易行,可作为刺蒺藜总皂苷的质量控制指标。

HPLC法同时测定蒺藜药材中3种成分的含量

目的:建立同时测定蒺藜中3种成分含量的方法,比较不同生长期、不同产地及不同药用部位的蒺藜中3种指标成分含量,探究采收时间及种植地区的科学性。方法:以蒺藜中3种有效成分,即槲皮素-3-O-β-吡喃葡萄糖基-(1→6)-吡喃葡萄糖苷、异鼠李素-3-O-β-D-吡喃葡萄糖基-(1→6)-吡喃葡萄糖苷、Terrestrinone A2为指标成分,高效液相色谱(HPLC)法测定含量。结果:3种指标成分含量均逐渐递增至6月并达到最大,随后逐渐降低;均表现为茎叶>果>根,根中含量低于检测限;产于内蒙古的蒺藜指标成分含量远远高于其他地区,其次为山东、河北、陕西。结论:该方法简便、准确,具有较好的重复性和稳定性,可为蒺藜质量控制提供参考。

18个不同产地蒺藜药材中蒺藜皂苷元的HPLC含量测定

目的采用HPLC法测定蒺藜中蒺藜皂苷元（25R-Spirostan-4-ene-3,12-dione）的含量。方法采用色谱柱Welch Ultimate LP-C18（250 mm×4.6 mm,5μm）;流动相：甲醇-水（80∶20）;检测波长：203 nm;柱温：30℃;流速：1.0 m L/min;进样量：20μL。结果蒺藜皂苷元对照品在0.82~7.38μg与峰面积呈良好的线性关系（r=0.999 7）,平均回收率为99.3%,RSD=1.67%（n=6）。结论所建立的方法灵敏、准确、重复性好,可用于蒺藜药材的含量测定。18个产地的蒺藜样品中以内蒙产的蒺藜皂苷元含量最高。