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.

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.

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.

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.

Protection by the gross saponins of Tribulus terrestris against cerebral ischemic injury in rats involves the NF-κB pathway

The aim of this study was to investigate whether the gross saponins of Tribulus terrestris(GSTT),a traditional Chinese herbal medicine,have neuroprotective effects on rats subjected to middle cerebral artery occlusion(MCAO),through nuclear factor-κB(NF-κB)pathway and inflammatory mediators.Cerebral ischemia was produced by MCAO in either untreated(control)or GSTT-pretreated rats,and the animals were examined for infarct volume,cerebral edema,neuro-behavioral abnormality and pathological changes.Meanwhile,the expression of NF-kB protein in brain tissue was analyzed on Western blots and the serum levels of TNF-α and IL-1 were determined by ELISA.The experimental results demonstrated that,compared with the control MCAO group,GSTT-pretreated MCAO group had significantly reduced infarct volume,brain edema and neuro-behavioral abnormality,and lesser degree of pathologic changes in the brain,as well as had lower levels of serum TNF-α and IL-1β,and higher levels of brain NF-κB(Po0.05).Furthermore,treatment with an NF-κB inhibitor pyrrolidine dithiocarbamate(PDTC)abolished the protective effects of GSTT against MCAO-induced cerebral ischemic injury.These results indicated that GSTT’s ability to protect against cerebral ischemic injury was mediated through the NF-κB signaling pathway,and that GSTT may act through inhibition of the production of inflammatory mediators.

基于PI3K-AKT信号通路探讨蒺藜皂苷对Graves眼病细胞凋亡的影响

目的探究蒺藜皂苷对Graves病体外细胞模型人甲状腺正常细胞Nthy-ori-3-1凋亡的影响及其分子机制。方法M22诱导人甲状腺滤泡上皮细胞株(Nthy-ori-3-1)建立Graves体外细胞模型,设对照组、模型组、蒺藜皂苷处理组(5、10、20 ng·mL^(-1))。CCK-8评估各组Nthy-ori-3-1细胞增殖能力;ELISA评估各组细胞培养液中cAMP变化;TUNEL评估各组细胞凋亡情况;免疫荧光评估各组细胞Bcl-2、Bax和Caspase-3变化;Western Blot评估PI3K、AKT、p-PI3K、p-AKT蛋白变化。结果高浓度蒺藜皂苷促进Nthy-ori-3-1细胞凋亡,抑制Nthy-ori-3-1细胞增殖;高浓度蒺藜皂苷抑制M22诱导Nthy-ori-3-1细胞cAMP表达;高浓度蒺藜皂苷促进Bax和Caspase-3表达,抑制Bcl-2表达;高浓度蒺藜皂苷抑制p-PI3K和p-AKT表达。结论高浓度蒺藜皂苷抑制PI3K-AKT信号通路促进Graves细胞凋亡。

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。

蒺藜化学成分的研究

目的：研究蒺藜（ Ｔｒｉｂｕｌｕｓｔｅｒｒｅｓｔｒｉｓ Ｌ．） 中生物活性物质。方法：用硅胶柱色谱、ＨＰＬＣ进行分离，根据光谱数据鉴定其结构。结果：分离纯化得到两种呋甾皂苷，确定其结构为２６ＯβＤ吡喃葡糖基（２５ Ｒ，Ｓ）５α呋甾１２羰基２０（２２）烯基３β，２６二醇３ＯβＤ吡喃葡糖基（１ →４）βＤ吡喃半乳糖苷（Ｉ） ；２６ＯβＤ吡喃葡糖基（２５ Ｒ）５α呋甾１２羰基３β，２２α，２６三醇３ＯβＤ吡喃葡糖基（１→２）βＤ吡喃半乳糖苷（ＩＩ）。结论：Ｉ。

注射用蒺藜皂苷的抗血栓形成作用

目的 :研究注射用蒺藜皂苷 (GSTT)对体内外血栓形成的影响。方法 :1采用动 -静脉旁路血栓形成法、电刺激动脉血栓形成法和体外血栓法 ,观察 GSTT对体内血栓湿重、血栓形成时间以及体外血栓长度和干、湿重的影响 ;2建立血瘀模型 ,观察 GSTT对血瘀大鼠全血高切、低切粘度的影响。结果 :1GSTT2 0 .8和31.2 mg· kg- 1可降低血栓湿重 (P<0 .0 5或 P<0 .0 1) ,延长血栓形成时间 (P<0 .0 1) ,缩短体外血栓长度(P<0 .0 5 ) ,降低体外血栓湿重和干重 (P<0 .0 1或 P<0 .0 5 ) ;2 GSTT2 0 .8和 31.2 m g· kg- 1可明显降低血液黏度 (P<0 .0 1)。结论 :GSTT具有抗血栓形成作用 ,其机制与降低血液黏度、改善血液流变性有关。

蒺藜皂苷对乳腺癌细胞Bcap-37的体外抑制作用

采用生长曲线法、MTT法、蛋白质含量测定以及形态学方法观察了蒺藜皂苷(STT)对于体外培养Bcap-37细胞抑制作用,结果显示:STT对于Bcap-37细胞增殖有较强的抑制作用,且呈明显的剂量与时间依赖关系,经STT作用后,细胞形态学表现为细胞变圆、皱缩,核固缩、浓聚。

蒺藜皂苷对大鼠小肠α-葡萄糖苷酶的抑制作用

目的:研究蒺藜皂苷(STT)对大鼠小肠α-葡萄糖苷酶的抑制作用及对餐后血糖水平的影响。方法:提取大鼠小肠α-葡萄糖苷酶,测定STT对其活性的抑制作用;一次性灌胃蔗糖(2 g.kg-1)或葡萄糖(2 g.kg-1),同时灌胃STT(100 mg.kg-1),测定60 min后血糖水平的变化;每天一次性灌胃STT(100 mg.kg-1),连续14 d,测定小肠α-葡萄糖苷酶活性以及灌胃蔗糖(2 g.kg-1)60 min后血糖水平的变化。结果:0.1,1,10 mg.mL-1的STT能够显著抑制α-葡萄糖苷酶的体外活性(P<0.001),抑制率分别为(20.83±1.66)%,(43.73±2.39)%和(52.62±2.69)%;STT显著抑制大鼠一次性灌胃蔗糖60 min后血糖水平的升高(P<0.01),变化幅度为对照的(52.61±6.24)%,但对灌胃葡萄糖没有作用;连续灌胃STT 14 d后,α-葡萄糖苷酶活性显著低于对照(P<0.05),为对照的(58.17±3.24)%,灌胃蔗糖60 min后血糖变化幅度为对照的(69.50±4.28)%(P<0.05)。结论:STT抑制大鼠餐后血糖水平的升高,是由其对小肠α-葡萄糖苷酶的抑制作用实现的。

天然产物 T12抗真菌作用及其机制研究

目的:研究从刺蒺藜(Tribulus terrestris L.)分离的一种皂苷类化合物T12的抗真菌作用及机制。方法:采用微量液基稀释法测定T12体外MIC_(80)值;透射电镜观察T12对白色念珠菌超微结构的影响;薄层色谱法、气质联用色谱法分别研究T12对细胞膜麦角甾醇生物合成通路的影响。结果:T12具有强的抗真菌活性,对白色念珠菌的体卟MIC_(80)值为0.87μmol·L^(-1);透射电镜观察,可见T12作用后,细胞膜多处破裂,细胞壁未见明显改变;T12对细抱膜麦角甾醇生物合成通路有明显影响。结论:结构新型、具有强抗菌活性的T12,通过影响白色念珠菌细胞膜麦角甾醇的生物合成发挥抗真菌作用。

蒺藜果中两种新甾体皂苷的分离和鉴定

目的 研究蒺藜 (TribulusterrestrisJ.)果实的化学成分。方法 用各种色谱技术进行分离和纯化 ,用ESI MS ,IR ,1 HNMR ,1 3 CNMR ,DEPT ,1 H 1 HCOSY ,1 H 1 3 CCOSY和HMBC等光谱技术鉴定结构。结果 从蒺藜果实中分得 2个新甾体皂苷 ,经光谱鉴定化合物I为新海柯皂苷元 3 O β D 吡喃葡糖基 (1→ 2 ) β D 吡喃葡糖基 (1→ 4) β D 吡喃半乳糖苷 ,化合物II为新海柯皂苷元 3 O β D 吡喃葡糖基 (1→ 4) β D 吡喃半乳糖苷。 结论 化合物I和II为新化合物 ,分别命名为蒺藜皂苷A和B。

蒺藜果总皂苷对大鼠实验性脑缺血的保护作用

目的 研究蒺藜果总皂苷对大鼠实验性脑缺血的保护作用。方法 采用结扎大鼠大脑中动脉的方法制备脑缺血模型 ,测定脑含水量及血管通透性 ,并观察脑缺血后血中内皮素含量、超氧化物歧化酶 (SOD)、6 - Keto-PGF1α和 TXB2 等指标。结果 蒺藜果总皂苷 2 0 ,5 0 ,10 0 mg/ kg均能明显减轻脑含水量及血管通透性 ,降低脑缺血后大鼠血中内皮素含量 ,提高血中 SOD活性。

蒺藜皂苷对预防小鼠高脂血症中肝脂酶和脂蛋白脂酶的作用及意义

目的:观察蒺藜皂苷(tribu saponin from Tribulus terrestris,STT)对预防小鼠高脂血症模型中脂蛋白脂酶(lipoprotein lipase,LPL)、肝脂酶(hepatic lipase,HL)活性的变化。方法:高脂膳食喂饲小鼠,并设正常对照、高脂对照、非诺贝特片阳性对照和STT高中低剂量组。测定血脂生化指标,同时检测血浆、肝组织中HL活性以及血浆、肝组织、脂肪组织和骨骼肌组织中LPL活性。结果:非诺贝特片及STT均能使肝组织中HL活性升高(P<0.01,P<0.05),STT能降低血浆中HL活性(P<0.05),而非诺贝特片组HL活性与高脂组无显著性差异(P>0.05)。肝组织中,非诺贝特片和STT能明显提高LPL活性(P<0.01,P<0.05);各组血浆中LPL活性均无明显改变(P>0.05);非诺贝特片和STT能提高骨骼肌组织中LPL活性(P<0.05),同时明显降低脂肪组织中LPL活性(P<0.01,P<0.05)和脂肪组织与肌肉组织中LPL的活性比值(P<0.01,P<0.05)。结论:STT能增加预防小鼠高脂血症模型肝组织中HL和LPL脂酶活性,降低脂肪组织与肌肉组织中LPL的活性比值,这一作用与降低血浆胆固醇水平和预防高脂血症密切相关。

蒺藜皂苷对大鼠离体心脏血液动力学的影响

目的 :观察蒺藜皂苷 (GSTT)对大鼠离体心脏血流动力学的影响。方法 :采用 L angendorff离体心脏灌流方法 ,以动脉血压 (AP)、左室收缩压 (L VSP)、左室舒末压 (L VEDP)、室内压最大上升速率 (+dp/ dtmax)、室内压最大下降速率 (- dp/ dtmax)及心率 (HR)、主动脉流量 (AF)、冠脉流量 (CF) 8项血流动力学参数为指标 ,观察 GSTT对心肌收缩性能的直接影响。结果 :给药后 5～ 2 0 min,10 0、15 0和 2 0 0 mg· L- 1 3个剂量组 GSTT均可使 AP、L VSP、± dp/ dtmax、AF的绝对值降低 ,使 L VEDP、CF的绝对值升高 ,与对照组比较差异均有显著性 (P<0 .0 5或 P<0 .0 1) ;GSTT3个剂量组均可拮抗因细胞外液钙离子浓度增加所致的正性肌力作用 ,同时拮抗因细胞外液葡萄糖浓度降低所致的心肌收缩力减弱 ,与对照组比较差异均有显著性 (P<0 .0 5或 P<0 .0 1)。结论 :GSTT可使离体工作心脏产生剂量依赖性的负性肌力作用 ,同时增加正常离体大鼠心脏冠脉流量 ,其负性肌力作用可能与阻滞钙通道有关 ,且这种负性肌力对心肌缺糖具有保护作用。

蒺藜皂苷抗抑郁作用及机制研究

目的探讨蒺藜皂苷的抗抑郁作用及机制。方法采用孤立大鼠加慢性轻度不可预见性应激方法造抑郁大鼠模型,进行开放旷场,悬尾行为学测试,用LC-MS/MS液质联用法检测大鼠血液中5-羟色胺(5-HT)、γ氨基丁酸(GABA)、去甲肾上腺素(NE)、谷氨酸(Glu)含量变化。实时定量PCR检测海马吲哚胺2,3-双加氧酶(IDO)、核转录因子-κB(NF-κB)、炎性细胞因子白介素1β(IL-1β)基因表达。Western blot检测海马中吲哚胺2,3-双加氧酶(IDO)、细胞因子NF-κB(p65)蛋白水平表达。结果与空白组比较,模型组小鼠旷场运动距离、站立次数和悬尾不动时间明显增加(P<0.05),血液中5-羟色胺明显下降(P<0.05),海马中IDO、NF-κB、IL-1β基因表达上调,IDO、NF-κB蛋白水平明显上调(P<0.05)。与模型组比较,蒺藜皂苷给药组能明显减少慢性应激导致大鼠旷场运动距离、站立次数和悬尾不动时间(P<0.05)。蒺藜皂苷给药组能明显提高大鼠血液中5-羟色胺含量(P<0.05),上调海马中抗炎因子IL-10基因表达,明显下调IDO、IL-1β基因表达(P<0.05),明显下调IDO蛋白表达水平(P<0.05)。结论蒺藜皂苷能明显改善动物行为学,具有抗抑郁作用,其作用可能与提高血液中5-羟色胺水平,下调色氨酸前体代谢酶吲哚胺2,3-双加氧酶(IDO)、IL-1β基因表达和降低吲哚胺2,3-双加氧酶(IDO)蛋白表达水平有关。

蒺藜全草中总皂苷总含量的测定

目的 :测定蒺藜全草中总皂苷的含量。方法 :将对照品蒺藜皂苷 FSA- 1和蒺藜总皂苷分别溶于硫酸 -甲醇 (7∶ 3)溶液中 ,于 6 0℃作用 15 m in后 ,用分光光度法测定蒺藜总皂苷的含量。结果 :在波长 319nm处有最大的吸收峰 ,浓度与吸收值之间有良好的线性关系。结论 :通过测定 6批蒺藜全草中总皂苷的含量 ,表明了本方法简单 ,重现性好 。