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.

不同温度及诱导方法对熊蜂王Bombus terrestris组建蜂群的影响

周年人工繁育熊蜂最困难的时候是年轻蜂王创建新的蜂群。我们在实验室条件下采用不同的温度梯度和诱导方法促进蜂王建立蜂群。B .terrestris在温度为 2 8～ 2 9℃ ,采用bbP诱导方法 ,生产效率最高 ,成群率在 5 0 %以上 ,在没有bbP的情况下 ,用 (YDQ +1btW )诱导方法成群率也在 30 %以上 ,符合工厂化生产的要求。

Antibacterial and antifungal activities of different parts of Tribulus terrestris L. growing in Iraq

Antimicrobial activity of organic and aqueous extracts from fruits, leaves and roots of Tribulus terrestris L., an Iraqi medicinal plant used as urinary anti-infective in folk medicine, was examined against I l species of pathogenic and non-pathogenic microorganisms： Staphylococcus aureus, Bacillus subtilis, Bacillus cereus, Corynebacterium diphtheriae, Escherichia coli, Proteus vulgaris, Serratia marcescens, Salmonella typhimurium, Klebsiella pneumoniae, Pseudomonas aeruginosa and Candida albicans using microdilution method in 96 multiwell microtiter plates. All the extracts from the different parts of the plant showed antimicrobial activity against most tested microorganisms. The most active extract against both Gram-negative and Gram-positive bacteria was ethanol extract from the fruits with a minimal inhibitory concentration （MIC） value of 0.15 mg/ml against B. subtilis, B. cereus, P. vulgaris and C. diphtheriae. In addition, the same extract from the same plant part demonstrated the strongest antifungal activity against C. albicans with an MIC value of 0.15 mg/ml.

Tribulus terrestris extracts alleviate muscle damage and promote anaerobic performance of trained male boxers and its mechanisms： Roles of androgen, IGF-1, and IGF binding protein-3

Purpose: To investigate the effects of Tribulus terrestris(TT) extracts on muscle mass, muscle damage, and anaerobic performances of trained male boxers and its mechanisms: roles of plasma androgen, insulin growth factor 1(IGF-1), and IGF-1 binding protein-3(IGFBP-3).Methods: Fifteen male boxers were divided into exercise group(E, n = 7) and exercise plus TT group(E + TT, n = 8). The 2 groups both undertook3-week high-intensity and 3-week high-volume trainings separated by a 4-week rest. TT extracts(1250 mg/day) were orally administered by boxers in E + TT group. TT extract compositions were detected by UHPLC–Q-TOF/MS. Before and at the end of the 2 trainings, muscle mass, anaerobic performance, and blood indicators were explored.Results: Compared with E group, decreases of plasma CK(1591.5 ± 909.6 U/L vs. 2719.9 ± 832.5 U/L) and IGFBP-3(3075.5 ± 1072.5 ng/m L vs. 3950.8 ± 479.3 ng/m L) as well as increases of mean power(MP, 459.4 ± 122.3 W vs. 434.6 ± 69.5 W) and MP/body weight(MP/BW, 7.5 ± 0.9 W/kg vs. 7.1 ± 1.1 W/kg) were detected in E + TT group after a high-intensity training. For high-volume training, reduction of IGFBP-3(2946.4 ± 974.1 ng/m L vs. 3632.7 ± 470.1 ng/m L) and increases of MP(508.7 ± 103.2 W vs. 477.8 ± 49.9 W) and MP/BW(8.2 ± 0.3 W/kg vs.7.5 ± 0.9 W/kg) were detected in E + TT group, compared with E group. Muscle mass, blood levels of testosterone, dihydrotestosterone(DHT),and IGF-1 were not signifiantly changed between the 2 groups.Conclusion: Taking 1250 mg capsules containing TT extracts did not change muscle mass and plasma levels of testosterone, DHT, and IGF-1 but significantly alleviated muscle damage and promoted anaerobic performance of trained male boxers, which may be related to the decrease of plasma IGFBP-3 rather than androgen in plasma.

Terresoxazine,A Novel Compound with Benzoxazine Skeleton from Tribulus terrestris

Terresoxazine, a novel benzoxazine derivative was isolated from the fruits of Tribulus terrestris. Its structure was determined as 7-hydroxy-3, 3a-dihydro-5H-pyrrolo-[1,2-a] [3,1] -benzoxazin-1(2H)-one, on the basis of the spectral techniques and X-ray crystallographic analysis.

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.

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.

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 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.

Use of a Natural Compound Made of Ecklonia bicyclis Seaweed, Tribulus terrestris and Water-Soluble Chitosan Oligosaccharide, in Male Sexual Asthenia with Mild or Mild-Moderate Erectile Dysfunction and Serum Testosterone Levels at the Lower Limit of Normal

Objectives: to evaluate the effectiveness of a natural compound made of Ecklonia bicyclis Seaweed, Tribulus terrestris and water-soluble chitosan oligosaccharide, in the male sexual asthenia with mild or mild-moderate erectile dysfunction and serum testosterone levels between 280 and 350 ng/dl. Materials and Methods: 84 male patients affected by reduced libido and serum testosterone levels at the lower limit of normal, were recruited. We have separated patients in three different age groups: group A (18 - 45 years), group B (45 - 59 years), group C (>60 years). All subjects answered the International index of erectile function questionnaire (IIEF-5) and underwent determination of serum total testosterone before and after 30 days of treatment. Results: Before treatment, the group A showed mean (± standard deviation) total testosterone 321.9 ± 19.2 ng/dl and mean IIEF-5 18.6 ± 1.97, in the group B it was 318.5 ± 18.1 ng/dl and 16.3 ± 2.66, and finally in the group C it was 305.4 ± 13.1 ng/dl and 14.2 ± 1.95 respectively. After treatment mean total testosterone and mean IIEF-5 were respectively: group A (448 ± 111.46 ng/dl and 21.84 ± 3.41);group B (453.8 ± 105.23 ng/dl and 20.4 ± 3.81);group C (385.8 ± 87.29 ng/dl and 16.7 ± 3.84). Conclusions: The treatment with Ecklonia bicyclis, Tribulus terrestris and water-soluble chitosan oligosaccharide might represent a safe and effective option on the improvement of libido and erectile function in man with testosterone level at the lower limit of normal.

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.

Chromatographic finger print analysis of anti-inflammatory active extract fractions of aerial parts of Tribulus terrestris by HPTLC technique

Objective:To develop HPTLC fingerprint profile of anti-inflammatory active extract fractions of Tribulus terrestris(family Zygophyllaceae).Methods:The anti-inflammatory activity was tested for the methanol and its fractions(chloroform,ethyl acetate,n-butanol and aqueous)and chloroform extract of Tribulus terrestris(aerial parts)by injecting different groups of rats(6 each)with carrageenan in hind paw and measuring the edema volume before and 1,2 and 3 h after carrageenan injection.Control group received saline i.p.The extracts treatment was injected i.p.in doses of 200 mg/kg 1 h before carrageenan administration.Indomethacin(30 mg/kg)was used as standard.HPTLC studies were carried out using CAMAG HPTLC system equipped with Linomat IV applicator.TLC scanner 3.Reprostar 3,CAMAG ADC 2 and WIN CATS-4 software for the active fractions of chloroform fraction of methanol extract.Results:The methanol extract showed good antiedematous effect with percentage of inhibition more than 72%,indicating its ability to inhibit the inflammatory mediators.The methanol extract was re-dissolved in 100 mL of distilled water and fractionated with chloroform,ethyl acetate and n-butanol.The four fractions(chloroform,ethyl acetate,n-butanol and aqueous)were subjected to anti-inflammatory activity.Chloroform fraction showed good anti-inflammatory activity at dose of 200 mg/kg.Chloroform fraction was then subjected to normal phase silica gel column chromatography and eluted with petroleum ether-chloroform,chloroform-ethyl acetate mixtures of increasing polarity which produced 15 fractions(F1-F15).Only fractions F1,F2,F4,F5,F7,F9,F11 and F14 were found to be active,hence these were analyzed with HPTLC to develop their finger print profile.These fractions showed different spots with different R_f values.Conclusions:The different chloroform fractions F1,F2,F4,F5,F7,F9,F11 and F14 revealed 4,7,7,8,9,7,7 and 6 major spots,respectively.The results obtained in this experiment strongly support and validate the traditional uses of this Sudanese medicinal plant.

Separation and Bio-activities of Spirostanol Saponin from Tribulus terrestris

Gross saponins of Tribulus terrestris（GSTT） have exact effect on cardiovascular and cerebrovascular diseases.But as a mixture,the specific efficient component of GSTT is still unknown.Nine monomers of spirostanol saponins were isolated and idendified as JA―JI（named transitorily） by means of NMR spectrometry.After bio-activity screening on them,we defined that monomers tigogenin 3-O-β-D-xylopyranosyl（1→2）-[β-D-xylopyranosyl（1→3）]-β-D-glucopyranosyl（1→4）-[α-L-rhamnopyranosyl（1→2）]-β-D-galactopyranoside（compound JB） and hecogenin3-O-β-D-glucopyranosyl（1→4）-β-D-galactopyranoside（compound JG） have cytoprotective bio-activity.Compound JB display effective dose in 10-8 and 10-9 mol/L,and JG in 10-6―10-9 mol/L.Survival rate,lactate dehydrogenase（LDH） and apoptosis also show that JB（at dose 10-8,10-9 and 10-10 mol/L） can protect myocardial injury caused by hypoxia/reoxygenation（H/R）.While morphology change also shows JG has cytoprotective bio-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.

Elastosonographic Changes in Patients with Peyronie's Disease, before and after Treatment with a Compound Based on <i>Ecklonia bicyclis</i>, <i>Tribulus terrestris</i>, and Water-Soluble Chitosan

Several diagnostic techniques have been proposed during the time for the diagnostic evaluation of Induratio Penis Plastica (IPP), due to poor sensitivity and specificity of standard B-Mode Ultrasonography. Shear-Wave elastosonography (SWE) is a relative new ultrasound technique, already used for several organs, which allows evaluating the rigidity of the tissues. Furthermore, we used a compound consisting of Ecklonia bicyclis, Tribulus terrestris, and Biovis (water-soluble Chitosan) with antifibrotic and antioxidant properties and analyzed the elastosonographic variations following the assumption of the compound. From March 2016 to April 2017, 40 patients were recruited with a mean age of 62. All patients underwent elastosonographic measurements at the start of treatment. Subsequently each patient took 1 tablet of the compound of Ecklonia bicyclis, Tribulus terrestris, and Biovis (water-soluble Chitosan) for 6 months. All patients underwent elastosonographic investigation at the end of treatment. The elastosonographic measurement prior treatment, showed that the average of the three measurements (proximal, medial and distal), was 37.05 KPa ± 7.76 SD and 38.8 KPa ± 10.11 SD for the left and right cavernous body, respectively. The examination at the end of the six months of treatment, with the compound showed that the average values of fibrosity were 31.07 KPa ± 7.5 SD and 30.86 KPa ± 11.4 SD for the left and right cavernous body respectively, demonstrating a decrease in fibrosity that was statistically significant (P < 0.01). Our data demonstrate that taking the compound for six months reduces the stiffness of the corpora cavernosa.

Protective Effects of Tribulus terrestris,Avena sativa,and White Ginseng Powder on Bone Mineral Density in Hypercholesterolemic Rats

This study investigated the effects of the herbal compounds Tribulus terrestris(TT),Avena sativa(AS),white ginseng(WG),and a triple combination(TC)powder on the serum total cholesterol,low-density lipoprotein cholesterol(LDL-C),high-density lipoprotein cholesterol(HDL-C),triglyceride,and the tibial bone mineral density(BMD)levels in rats fed a high-cholesterol diet.This study comprised Group I(normal pellet feed),Group II(pellet feed with 2%cholesterol),Group III(2%cholesterol plus 15%TT),Group IV(2%cholesterol plus 7.5%AS),Group V(2%cholesterol plus 5%WG)and Group VI(2%cholesterol plus 7.5 TT%+3.75%AS,2.5%WG).The serum total cholesterol,LDL-C,HDL-C,triglyceride,and tibial BMD(g/cm2)levels were measured.Significant decrease in the serum HDL-C levels in Group II than Group I,and significant increase in the serum LDL-C levels in Group II than Groups I and VI were determined.The tibial BMD levels were significantly lower in Groups II and IV than Group I.It was determined that WG and TC significantly prevented the serum total cholesterol increase;TC significantly prevented the serum LDL-C increase;and TT,WG,and TC were non-significantly effective in the improvement of tibial BMD levels.

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.

Lactobacillus melliventris promotes hive productivity and immune functionality in Bombus terrestris performance in the greenhouse

Bumblebees are important pollinators in agricultural ecosystems,but their abundance is declining globally.There is an urgent need to protect bumblebee health and their pollination services.Bumblebees possess specialized gut microbiota with potential to be used as probiotics to help defend at-risk bumblebee populations.However,evidence for probiotic benefits on bumblebees is lacking.Here,we evaluated how supplementation with Lactobacillus melliventris isolated from bumblebee gut affected the colony development of Bombus terrestris.This native strain colonized robustly and persisted long-term in bumblebees,leading to a significantly higher quality of offspring.Subsequently,the tyrosine pathway was upregulated in the brain and fat body,while the Wnt and mTOR pathways of the gut were downregulated.Notably,the field experiment in the greenhouse revealed the supplementation of L.melliventris led to a 2.5-fold increase in the bumblebee survival rate and a more than 10%increase in the number of flowers visited,indicating a better health condition and pollination ability in field conditions.Our study represents a first screening for the potential use of the native gut member,L.melliventris,as probiotic strains in hive supplement for bumblebee breeding,which may be a practical approach to improve immunity and hive health.

What is the effect of Tribulus terrestris on testicular morphology and sperm production?An experimental study in the rat model

The objective of this study was to investigate whether treatment with Tribulus terrestris(Tt)has any impact on the testicular morphology and function in a rodent model.Twenty male rats were divided into a control group and a group receiving 100 mg kg^(-1) body weight of Tt supplementation.After 4o days of experiment,the animals were submitted to euthanasia;epididymal tail spermatozoa were collected;and spermatozoa concentration,motility,and viability were analyzed.In addition,testicles were collected and processed for histomorphometrical analyses.Data were compared using the Student's t-test and considered significant when P<O.05.Spermatozoa concentration,motility,and viability showed no difference between the groups.Further,testicular weight and volume,seminiferous tubule diameter,tunica propria surface density,seminiferous epithelium surface density,and intertubular compartment surface density were statistically similar between the groups.However,seminiferous epithelium height and tubular lumen surface density were augmented in animals treated with Tt.Treatment with Tt does not cause a major impact on testicular morphology,promoting only subtle modifications.No difference on spermatozoa parameters was observed.