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.

Two new furostanol saponins from Tribulus terrestris L.

Two new furostanol glycoside,26-O-β-D-glucopyranosyl-(25S)-5α-furost-3β,22α,26-triol-3-O-β-D-rhamnopyranosyl-(1→2)-[β-D-glucopyranosyl-(1→4)]-β-D-galactopyranoside(1) and 26-O-β-D-glucopyranosyl-(25S)-5α-furost-20(22)-en-2α,3β,26-triol-3-O-β-D-glucopyranosyl-(1→2)-O-β-D-glucopyranosyl-(1→4)-β-D-galactopyranoside(2) were isolated fromthe fruits of Tribulus terrestris L.The structures of two new furostanol saponins were elucidated by spectroscopic methods.

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.

Effects of Tribulus terrestris saponins on proliferation and invasion of A549 cells

Objective:Tribulus terrestris saponin is a traditional Chinese medicine in China.This experiment was designed to investigate the effects of tribulus terrestris saponin on the proliferation and invasion ability of non-small cell lung cancer A549 cells.Methods:A549 cells were divided into normal control and experimental groups(Tribulus terrestris saponin 250μg/mL group,Tribulus terrestris saponin 200μg/mL group,Tribulus terrestris saponin 150μg/mL group,Tribulus terrestris saponin 100μg/mL group,Tribulus terrestris saponin 50μg/mL group).The proliferation viability of the cells in each group was detected by CCK8,the invasion of tumor cells was detected by Transwell model.The mRNA expression of MMP9 and caspase-3 in each group of cells was detected by RT-PCR.Immunofluorescence staining was used to observe the fluorescence intensity of caspase-3 in each group of cells.Results:Compared with the normal control group,tribulus terrestris saponin significantly inhibited the proliferation activity and invasion ability of A549 cells,which was statistically significant(P<0.01).In the invasion assay,compared with the control group,MMP9 expression was significantly reduced and caspase-3 expression was significantly increased in the tribulus terrestris saponin group,and both were concentration-dependent,with statistically significant differences(P<0.01).By cellular immunofluorescence staining experiments,it was found that the fluorescence expression of caspase-3 was enhanced in the experimental group compared with the normal control group,in which the high concentration saponin group was significantly higher than the low concentration group.Conclusion:Tribulus terrestris saponin can inhibit the invasive ability of A549 cells by down-regulating the expression of MMP9,and induce irreversible apoptosis by up-regulating the activation of caspase-3 expression to form caspase-3.

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.

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.

Synergistic Hepatoprotective Effect of Feronia fimonia L,, Citrullus colocynthis L,, and Tribulus terrestris L, against Paracetamol Induced Hepatotoxicity in Swiss Albino Rats

Paracetamol toxicity induced a significant rise in AST （aspartate transaminase）, ALT （alanine transaminase）, ALP （alkhaline phisphatase）, total bilirubin, total cholesterol, Triglycerides. Administration of 150 mg/kg b.w of synergistic ethanolic extracts of Feronia limonia L., Citrullus colocynthis L., and Tribulus terrestris L. effectively reduced these pathological damages caused by paracetamol intoxication. In addition to serum parameters treatment of 150 rng/kg b.w of synergistic ethanolic extract of Feronia limonia L., Citrullus colocynthis L. and Tribulus terrestris L. also promote the body weight in albino rats. Histopathological changes of the liver samples were compared with the normal control. From our results, we may infer that the mode of action of synergistic 90% ethanolic extracts of Feronia limonia L., Citrullus colocynthis L., and Tribulus terrestris L. （150 mg/kg b.w） in affording the in vivo hepatoprotective activity against paracetamol may be due to the cell membrane stabilization, hepatic cell regeneration and activation of antioxidative enzymes such as glutathione reductase, glutathione peroxidase, superoxide dismutase and catalase.

HPLC-CAD法与ELSD法用于测定蒺藜皂苷提取物中蒺藜皂苷K的对比研究

目的:分别建立高效液相色谱-电雾式(CAD)和蒸发光散射(ELSD)2种通用型检测器测定蒺藜皂苷提取物中蒺藜皂苷K含量的方法。方法:采用高效液相色谱法,Waters Symmetry Shield RP18为色谱柱,以乙腈-水为流动相,梯度洗脱,体积流速为1.0 mL·min-1,柱温为25℃;使用CAD和ELSD检测器测定,并对样品实测结果进行比较。结果:2种方法线性关系均良好,精密度、重复性、稳定性及加样回收率皆符合方法学要求,CAD的检出限和定量限均比ELSD低。结论:2种检测方法都可用于蒺藜皂苷提取物中蒺藜皂苷K的含量测定,CAD检测器比ELSD检测器具有更高的灵敏度和精密度。

基于TLR4/NF-κB通路研究蒺藜总皂苷对周围神经病理性疼痛的镇痛作用

目的:基于TLR4/NF-κB通路研究蒺藜总皂苷(gross saponins from tribulus terrestris,GSTT)对大鼠慢性坐骨神经损伤(chronic constriction injury,CCI)所引起的周围神经病理性疼痛是否具有镇痛作用。方法:选取雄性SD大鼠40只,随机分为假手术组(SHAM组)、CCI组、GSTT 100 mg/kg+CCI组、GSTT 200 mg/kg+CCI组、普瑞巴林+CCI组(阳性对照)5组,每组8只。除SHAM组,其余各组均制备CCI大鼠模型,分别以药物处理后,检测各组大鼠机械缩足反射阈值(paw withdrawal mechanical threshold,PWMT)、热敏潜伏期;以HE染色检测各组大鼠脊髓及背根神经节组织病理形态变化;免疫组化检测脊髓及背根神经节组织中NF-κB的表达。结果:与SHAM组相比,CCI组大鼠PWMT、热敏潜伏期明显降低(P<0.05);损伤侧脊髓背角及背根神经节的神经元、胶质细胞染色较深,并出现肿胀、萎缩等损伤,细胞核浓缩、丢失;损伤侧脊髓背角及背根神经节中NF-κB表达水平明显增高(P<0.05)。与CCI组相比,GSTT 100 mg/kg+CCI组、200 mg/kg+CCI组,普瑞巴林+CCI组大鼠PWMT、热敏潜伏期升高(P<0.05),上述病理变化也明显改善,损伤侧脊髓背角及背根神经节中NF-κB水平降低。结论:GSTT对CCI大鼠引起的周围神经病理性疼痛具有镇痛作用,可能与TLR4/NF-κB通路有关。

蒺藜皂苷对人肝癌细胞BEL7402生长抑制和诱导凋亡作用的研究

目的 :探讨蒺藜皂苷 (STT)对体外培养的人肝癌细胞BEL 74 0 2的增殖抑制作用及诱导凋亡作用。方法 :应用MTT与SRB法检测蒺藜皂苷对人肝癌细胞BEL 74 0 2的抑制作用 ,采用形态学观察以及流式细胞术 (FCM)检测了STT诱导人肝癌细胞BEL 74 0 2凋亡的效应 ,采用流式细胞术检测BEL 74 0 2细胞Bcl 2蛋白表达。结果 :①STT能够剂量依赖地抑制人肝癌细胞BEL 74 0 2的增殖 ;②细胞显现典型的凋亡形态学改变 ,FCM出现凋亡峰 ;③STT能降低BEL 74 0 2细胞的Bcl 2蛋白表达。结论 :STT在体外能抑制BEL 74 0 2细胞的增殖并能诱导细胞凋亡 ,其诱导凋亡作用的途径之一是下调Bcl 2蛋白表达。

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。

蒺藜总皂苷对心肌缺血再灌注损伤炎症因子TNF-α、IL-1β释放的影响

目的观察蒺藜总皂苷(GSTT)对大鼠心肌缺血再灌注损伤(MI/R)炎症因子TNF-α、IL-1β释放的影响。方法可逆性冠脉左前降支结扎缺血45min再灌注2h复制MI/R模型,入选SD雄性大鼠32只,随机分成假手术组、模型组、GSTT大剂量组、GSTT小剂量组,每组8只。放免法检测血清TNF-α、IL-1β含量。结果GSTT大剂量组TNF-α、IL-1β含量显著降低(P<0·05)。结论GSTT对MI/R有保护作用,可抑制炎症损伤,减少炎性因子TNF-α、IL-1β的释放。

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。

蒺藜总皂苷对缺氧复氧损伤心肌细胞凋亡及凋亡相关基因bcl-2与bax表达的影响

目的观察蒺藜总皂苷(GSTT)对培养乳鼠心肌细胞模拟缺血再灌注损伤后心肌细胞凋亡及凋亡相关基因bcl-2、bax表达的影响。方法利用原代培养的SD乳鼠心肌细胞建立模拟心肌缺血再灌注(MI/R)模型,分正常培养组、模型组和蒺藜总皂苷大、小剂量组。流式细胞仪检测心肌细胞凋亡率,免疫组化检测心肌细胞凋亡基因bcl-2、bax的表达。结果GSTT大、小剂量组均可显著降低细胞凋亡率(P<0·05)。GSTT大剂量组可显著上调bcl-2表达(P<0·01)、下调bax表达(P<0·05),而GSTT小剂量组可上调bcl-2表达(P<0·05),但对bax表达无明显影响。结论GSTT可抑制模拟缺血再灌注损伤心肌细胞的凋亡,其作用可能是通过上调bcl-2、下调bax表达实现的。

蒺藜皂苷提取工艺及其药理活性研究进展

蒺藜皂苷是中药材蒺藜的主要成分,其药理活性显著,具有抗癌、抗氧化、抗炎、抗菌、抗抑郁、抗疲劳、降血糖、保护心脑血管和保护神经等功效.综述了蒺藜皂苷的提取方法和药理活性,以期为蒺藜皂苷的深加工综合利用提供理论参考.

RP-HPLC蒸发光检测法测定蒺藜中蒺藜皂苷D的含量

目的：建立反相高效液相色谱-蒸发光检测法测定蒺藜皂苷D含量的方法。方法：色谱柱Nova-park C18柱,柱温22°C,流动相：甲醇-水（75∶25）为流动相,流速1 mL/min,SEDEX 55型蒸发光散射检测器。结果：该方法的线性范围为2.736-5.13μg,r=0.9994（n=5）,平均回收率为96.86%。结论：方法准确,灵敏度高,专属性强,重现性好。可用于该药材及制剂中该成分的含量测定。

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。