Antitumor activity of Bulgarian herb Tribulus terrestris L. on human breast cancer cells

Medicinal plants have been intensively studied as a source of antitumor compounds. Due to the beneficial climate conditions Bulgarian herbs have high pharmacological potential. Currently, the antitumor effect of the Bulgarian medicinal plant Tribulus terrestris L. on human cancer cell lines is not studied. The main active compounds of the plant are the steroid saponins.The present study aims to analyze the effect on cell viability and apoptotic activity of total extract and saponin fraction of Bulgarian Tribulus terrestris L. on human breast cancer (MCF7) and normal (MCF10A) cell lines. Antitumor effect was established by МТТ cell viability assay and assessment of apoptotic potential was done through analysis of genomic integrity (DNA fragmentation assay) and analysis of morphological cell changes (Fluorescence microscopy). The results showed that total extract of the herb has a marked dose-dependent inhibitory effect on viability of MCF7 cells (half maximal inhibitory concentration is 15 μg/ml). Cell viability of MCF10A was moderately decreased without visible dose-dependent effect. The saponin fraction has increased inhibitory effect on breast cancer cells compared to total extract. Morphological changes and DNA fragmentation were observed as markers for early and late apoptosis predominantly in tumor cells after treatment. Apoptotic processes were intensified with the increase of treatment duration.The obtained results are the first showing selective antitumor activity of Bulgarian Tribulus terrestris L. on human cancer cells in vitro. Apoptotic processes are involved in the antitumor mechanisms induced by the herb. This results give directions for future investigations concerning detailed assessment of its pharmacological potential

ПРОУЧВАНЕ ВЛИЯНИЕТО НА СУХ ЕКСТРАКТ ОТ TRIBULUS TERRESTRIS ВЪРХУ ОСНОВНИ БИОХИМИЧНИ И ХЕМАТОЛОГИЧНИ ПОКАЗАТЕЛИ НА КРЪВТА ПРИ ТОКАЧКИ (NUMIDA MELEAGRIS)

The aim of the present research was to investigate the effect of Bulgarian additive Vemoherb-T (dry extract of the annual plant Tribulus terrestris – L), produced by Vemo 99 Ltd Company, Sofia on main biochemical characteristics and hematological parameters of the blood in guinea fowl (Numida meleagris). An experiment was carried out with 30 Pearl-gray Guinea fowl (32 weeks old), distributed in two groups – a control and an experimental, 12 female and 3 male each. All birds were fed the same mixture for breeder guinea fowl. Vemoherb-T was supplemented to the compound feed of the experimental group in a daily dose of 10 mg/kg body weight for a period of 12 weeks. The tested product decreased significantly the levels of total triglycerides (P < 0.05), total cholesterol (P <0.01) and glucose (P < 0.01; P < 0.001 in male and female birds respectively) in the blood serum. It was established significantly higher total protein- ((P < 0.001) and calcium (P<0.01) values in the blood serum of the treated birds. The addition of Vemoherb-T increased significantly hemoglobin level, the number of erythrocytes and leukocytes and decreased the number of eosinophils in guinea fowl from the both sexes.Целта на настоящото изследване беше да се установи влиянието на Българския продукт Vemoherb-T (сух екстракт от едногодишното растение Tribulus terrestris-L), произведен от фирма Вемо 99 ООД, София, върху основни биохимични характеристики и хематологични показатели на кръвен серум при токачки (Numida meleagris). Беше проведен научен експеримент с 30 Бисерносиви токачки (на 32 седмична възраст), разпределени в две групи – опитна и контролна, 12 женски и 3 мъжки всяка. Всички птици получаваха една и съща смеска за разплодни токачки. Vemoherb-T беше добавян към комбинирания фураж на опитната група ежедневно в доза 10 mg/kg жива маса в продължение на 12 седмици. Изпитваният продукт понижава достоверно нивата на общите триглицериди (P < 0.05), общия холестерол (P <0.01) и глюкозата (P < 0.01; P < 0.001 съответно за мъжките и женски птици) в кръвния серум. Бяха установени достоверно по- високи стойности на общия протеин ((P < 0.001) и на калция (P < 0.01) в кръвния серум на третираните птици. Добавката на Vemoherb-T повишава достоверно нивото на хемоглобина, броя на еритроцитите и лeвкоцитите, и понижава броя на еозинофилните левкоцити при токачките и от двата пола

Effectiveness of Active Compounds of Herbal Plants as Aphrodisiacs Through Molecular Docking Against Human Phosphodiesterase-5 Receptors

Based on the evaluation of side effects the use of sildenafil as a human phosphodiesterase 5 inhibitor drug (HPDE5) has prompted the search for new compounds that have the potential to be aphrodisiacs. The purpose of this study was to determine the interaction of the active compounds nilocitin, stigmasterol, protodioscin, icariin, yohimbine, and ginsenoside against the HPDE5 receptor as an aphrodisiac. The method used in this study was experimental conducted in silico. The metabolite structure is downloaded from the PubChem application, the protein is downloaded from PDB (Protein Data Bank) with the code 2H42. The result of this study is that the active compound may interact with HPDE5 receptors. The interaction that occurs results in the formation of van der waals bonds, hydrogen, carbon hydrogen, sigma, sulfur cation anions, T-shape and alkyls. The active compounds each have a sildenafil bond energy of -9.5 kcal/mol; niloticin -7.8 kcal/mol; stigmasterol -10.7 kcal/mol; protodioscin -13.1 kcal/mol; icariin -11.1 kcal/mol; yohimbine -10.1 kcal/mol and ginsenoside -12.1 kcal/mol with RMSD 0. The interaction with the HPDE5 receptor results in the formation of the same amino acid residue as the comparison ligand. The residual equation shows that the compounds have the same activity and can be predicted as aphrodisiac

ACUTE TOXICITY EVALUATION OF PROTODIOSCIN RICH EXTRACT OF TRIGONELLA FOENUM-GRAECUM L IN RATS

ABSTRACTObjectives: The objective of this study was to investigate the acute toxicity of standardized protodioscin rich extract (PRE) of Trigonella foenumgraecumL (26.63% w/w; PRE).Methods: To evaluate the toxicity of PRE, the acute toxicity of the extract on adult rats were investigated. A fixed large dose of 2 g/kg body weight ofPRE was administrated by a single oral gavage, and 1 g/kg body weight of PRE was administered by intravenous according to the Organisation forEconomic Co-operation and Development guidelines.Results: In 2 weeks, PRE showed no obvious acute toxicity. There were no deaths in either group and no change in the clinical signs. The hematologicaland biochemical analyses showed some changes that returned to reference levels without impairment of homeostasis. The treatment did not induceuntoward changes in organs as shown by histological studies. The in vivo results showed that has low toxicity.Conclusion: PRE is safe and can be potentially used as an aphrodisiac in future studies.Keywords: Protodioscin, Aphrodisiac, Trigonella foenum-graecum, Organisation for Economic Co-operation and Development, Toxicity,Histopathological studies

Bioactive Compounds, Antioxidant, Anti-Inflammatory, Anti-Cancer, and Toxicity Assessment of Tribulus terrestris—In Vitro and In Vivo Studies

Tribulus terrestris L. belongs to the family Zygophyllaceae and integral part of various ancient medicinal systems including Chinese, Indian, and European to combat various health ailments. The aim of the present study was to assess the phytochemical constituents, in vitro antioxidant activity using DPPH, FRAP, and H2O2 assays, in vitro anticancer activity using MTT assay, and in vitro and in vivo anti-inflammatory properties of T. terrestris. The acute and sub-acute toxicity of extracts exhibiting most biological potential was examined using murine models. Liquid–liquid partitioning followed by RP–HPLC sub-fraction of crude extract was performed. After that, ESI-MS/MS analysis was done for the timid identification of bioactive metabolites responsible for bioactivities of sub-fractions and HPLC analysis to quantify the compounds using external standards. Among all extracts, T. terrestris methanol extract was noted to hold maximum phenolic (341.3 mg GAE/g) and flavonoid (209 mg QE/g) contents, antioxidant activity in DPPH (IC50 71.4 µg/mL), FRAP (35.3 mmol/g), and H2O2 (65.3% inhibition) assays, anti-inflammatory activities in vitro at 400 µg/mL (heat-induced hemolysis, % inhibition 68.5; egg albumin denaturation, % inhibition 75.6%; serum albumin denaturation, % inhibition 80.2), and in vivo at 200 mg/kg (carrageenan-induced paw edema, % inhibition 69.3%; formaldehyde-induced paw edema, % inhibition 71.3%) and anti-cancer activity against breast cancer cell (MCF-7) proliferation (IC50 74.1 µg/mL). Acute and subacute toxicity studies recorded with no change in body weight, behavior, hematological, serum, and histopathological parameters in treated rats with T. terrestris methanol extracts when compared to control group. Fraction B obtained through liquid–liquid partitioning resulted in more bioactive potential as compared to the parent methanol extract. RP–HPLC analysis of fraction B resulted with four sub-fractions (TBTMF1-TBTMF4), wherein TBTMF3 delineated notable bioactive capabilities as compared to other fractions and parent methanol extract. ESI-MS/MS analysis of TBTMF3 resulted with tentative identification of myricetin, rutin, liquitrigenin, physcion, and protodioscin. It can be stated that T. terrestris is a potential bearing herb and findings of current study further verify the claims made in ancient medicinal systems. However, after investigation of each identified compound, it must be considered for drug discovery. © 2022 by the authors. Licensee MDPI, Basel, Switzerland

The disposition and pharmacokinetics of Dioscorea nipponica Makino extract in rats

This study was aimed to investigate the disposition and pharmacokinetics of the total saponins of dioscorea (TSD) in rats. Male Sprague-Dawley rats were orally administrated with 3H labeled TSD at a single dose ratio of 80 mg TSD per 1 kg rat. Blood samples and feces were collected at different time points to measure the level of TSD activity. At the final time point, determination of the disposition of TSD in lung, kidney, heart, liver, adrenal, and small intestine were performed. From the blood samples' emission of radioactivity, pharmacokinetic parameters were derived as T1/2 = 33.33 ± 4.48 h, T max = 6.5 ± 0.71 h, AUC = 119400 ± 421097.67, and C max = 2643.33 ± 192.26 dpm/ml. There was 51.609% of 3H labeled substance excreted in 24 h. These results suggested that blood concentration of 3H-TSD was extremely low and the majority of TSD was excreted in the feces. The TSD was extensively distributed to multitissues. The radioactivity level was measured to be the highest in the liver, adrenal gland, and wall of the gastrointestinal tract. The radioactivity of TSD was still being detected in blood after 96 h. This showed TSD was excreted in vivo very slowly. © 2008 Academic Journals.published_or_final_versio

Signal Grass (Brachiaria decumbens) Toxicity in Grazing Ruminants

Signal grass (Brachiaria decumbens) is a highly productive tropical grass that is widespread through South America, Australia, Indonesia, Vanuatu and Malaysia due to its adaptation to a wide range of soil types and environments. Animal production from these B. decumbens pastures is highly variable due to sporadic outbreaks of photosensitisation associated with low growth rates of young animals, anorexia and wasting. The identification of B. decumbens toxicity through clinical signs may grossly underestimate the impact and severity of the disease. Affected animals without clinical signs have elevated serum liver enzyme concentrations resulting from blockage of the bile ducts by birefringent crystals, identified as calcium salts of steroidal saponins found in leaves and stems. The concentrations of the steroidal saponins vary through the year and within the plant. Young, green leaves contain 5–10 times the saponin concentration of mature leaves indicating that B. decumbens pastures are likely to be more toxic during sprouting and early growth. Previous exposure, selective grazing, and avoiding toxic leaves may partly explain apparent resistance of some animals to B. decumbens toxicity. Further research is needed to define growing conditions that produce elevated saponin levels and to investigate the impact of B. decumbens on rumen function

Pengaruh Jamu Dengan Tribulus Terrestris Terhadap Kualitas Sperma Tikus Wistar Jantan (Rattus Norvegicus)

: It is known that Tribulus terrestris can improve sperm quality. One of the Indonesian traditional medicine, jamu, contains this Tribulus terrestris. This study aimed to investigate the effect of jamu containing Tribulus terrestris on sperm quality which consisted of concentration, motility, and morphology of spermatozoa of male wistar rats (Rattus norvegicus). This was an experimental study. Nine male wistar rats were randomly divided into 3 groups of 3 rats each: Group P0 (control), group P1 (given 5 mg of Tribulus terrestris), and group P2 (given 10 mg of Tribulus terrestris). This study was conducted for 52 days. The results showed that the spermatozoa concentration of P1 was 49x106 sperm/ml suspension and of P2 53x106 sperm/ml suspension; both were lower than of P0 (59x106 sperm/ml suspension). The sperm motility category (A+B) of P1 was 33% and of P2 (37%); both were lower than of P0 (45%). The motility category A was not found in P1 and P2. The percentage of spermatozoa with normal morphology of P1 was 45% and the abnormal ones was 55%, meanwhile in P2 there was 49% of normal sperm morphology and 51% of abnormal morphology. Conclusion: Jamu containing Tribulus terrestris had no effect in improvement of the sperm quality

Water-soluble saponins accumulate in drought-stressed switchgrass and may inhibit yeast growth during bioethanol production

Background: Developing economically viable pathways to produce renewable energy has become an important research theme in recent years. Lignocellulosic biomass is a promising feedstock that can be converted into second-generation biofuels and bioproducts. Global warming has adversely affected climate change causing many environmental changes that have impacted earth surface temperature and rainfall patterns. Recent research has shown that environmental growth conditions altered the composition of drought-stressed switchgrass and directly influenced the extent of biomass conversion to fuels by completely inhibiting yeast growth during fermentation. Our goal in this project was to find a way to overcome the microbial inhibition and characterize specific compounds that led to this inhibition. Additionally, we also determined if these microbial inhibitors were plant-generated compounds, by-products of the pretreatment process, or a combination of both. Results: Switchgrass harvested in drought (2012) and non-drought (2010) years were pretreated using Ammonia Fiber Expansion (AFEX). Untreated and AFEX processed samples were then extracted using solvents (i.e., water, ethanol, and ethyl acetate) to selectively remove potential inhibitory compounds and determine whether pretreatment affects the inhibition. High solids loading enzymatic hydrolysis was performed on all samples, followed by fermentation using engineered Saccharomyces cerevisiae. Fermentation rate, cell growth, sugar consumption, and ethanol production were used to evaluate fermentation performance. We found that water extraction of drought-year switchgrass before AFEX pretreatment reduced the inhibition of yeast fermentation. The extracts were analyzed using liquid chromatography–mass spectrometry (LC–MS) to detect compounds enriched in the extracted fractions. Saponins, a class of plant-generated triterpene or steroidal glycosides, were found to be significantly more abundant in the water extracts from drought-year (inhibitory) switchgrass. The inhibitory nature of the saponins in switchgrass hydrolysate was validated by spiking commercially available saponin standard (protodioscin) in non-inhibitory switchgrass hydrolysate harvested in normal year. Conclusions: Adding a water extraction step prior to AFEX-pretreatment of drought-stressed switchgrass effectively overcame inhibition of yeast growth during bioethanol production. Saponins appear to be generated by the plant as a response to drought as they were significantly more abundant in the drought-stressed switchgrass water extracts and may contribute toward yeast inhibition in drought-stressed switchgrass hydrolysates