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Objective - Diabetic hepatotoxicity involves complex events which include kupffer cell activation, formation of reactive oxygen species, cytokines release (TNF-α, IL-1β), and finally leads to hepatocyte death. “β- Aescin showed anti-inflammatory, anti-oxidant, gastroprotective and anti-oedema properties. The present study investigated the protective effect of β- Aescin in streptozotocin induced diabetic hepatotoxicity. Method - Female mice were divided into six groups, the first group served as the control, the second to sixth group received single i.p. dose of 90 mg/kg of STZ, the second group served as the untreated diabetic group, the third, fourth and fifth group received β- aescin intra-peritoneally at the dose of 0.9 mg/kg, 1.8 mg/kg and 3.6 mg/kg body weight respectively. The last sixth group was treated with 10 mg/kg glibenclamide i.p. for 14 days. A significant decrease in the blood glucose level was showed in β-aescin group as compared to the control group. Result - A significant increase of blood glucose level was observed in high and mid dose of β- aescin (3.6 mg/kg and 1.8 mg/kg respectively), standard drug (glibenclamide 10 mg/kg) groups as compared to control group. ROS generation was evaluated by using DCF-DA estimation method for the acute toxicity in liver tissue. Streptozotocin group showed more ROS generation in comparison to β- aescin group (3.6 mg/kg). Serum biochemical markers showed a significant decrease in β- aescin treated diabetic mice compared to untreated diabetic mice. Histopathological evaluation showed severe changes in untreated diabetic liver tissue marked by large number of inflammatory cells such as lymphocytes along with hepatic sinusoidal inflammation and hepatocyte necrosis whereas treated diabetic mice with β- aescin showed reduction in hepatotoxicity marked by regeneration changes of hepatocytes and mildly hepatocyte degeneration. Conclusion - In the study, β- aescin showed beneficial effects on the efficient properties of the liver and microscopic improvements in diabetic hepatotoxicity.

Ischemic stroke is one of the major causes of disability; widely use of endovascular thrombectomy or intravenous thrombolysis leads to more attention on ischemia-reperfusion injury (I/R injury). Aescin, a natural compound isolated from the seed of the horse chestnut, has been demonstrated anti-inflammatory and antiedematous effects previously. This study was aimed at determining whether aescin could induce protective effects against ischemia-reperfusion injury and exploring the underlying mechanisms in vitro. Primary cultured neurons were subjected to 2 hours of oxygen-glucose deprivation (OGD) followed by 24 hours of simulated reperfusion. Aescin, which worked in a dose-dependent manner, could significantly attenuate neuronal death and reduce lactate dehydrogenase (LDH) release after OGD and simulated reperfusion. Aescin treatment at a concentration of 50 μg/ml provided protection with fewer side effects. Results showed that aescin upregulated the phosphorylation level of PRAS40 and proteins in the mTOR signaling pathway, including S6K and 4E-BP1. However, PRAS40 knockdown or rapamycin treatment was able to undermine and even abolish the protective effects of aescin; meanwhile, the levels of phosphorylation PRAS40 and proteins in the mTOR signaling pathway were obviously decreased. Hence, our study demonstrated that aescin provided neuronal protective effects against I/R injury through the PRAS40/mTOR signaling pathway in vitro. These results might contribute to the potential clinical application of aescin and provide a therapeutic target on subsequent cerebral I/R injury.

In the present study, we show how acrylamide-based microgels can be employed for the uptake and release of the drug β-aescin, a widely used natural product with a variety of pharmacological effects. We show how aescin is incorporated into the microgel particles. It has an important influence on the structure of the microgels, by reducing their natural network-density gradient in the swollen state. Moreover, temperature-dependent measurements reveal how the incorporation of aescin stabilizes the microgel particles, while the volume phase transition temperature (VPTT) is almost constant, which is very important for the intended drug release. Finally, it is shown that upon increase of the temperature above the VPTT the particles are able to release aescin from their network, encouraging the use of this particular drug delivery system for hypothermia treatments., The authors acknowledge financial support by the Deutsche Forschungsgemeinschaft (DFG) grant number HE2995/7-1.

β - escin is a mixture of triterpene saponins and other components including alpha aescin, progestoescigenin, barringtogenol, cryptoescin and benzyopyrones.Βaescin or β-escin isolated from the horse chestnut seeds (Aesculus hippocastanum L.) β-escin has been traditionally used to treat conditons such as chronic venous insufficiency, inflammation, hemorrhoids, edema, elevated glucose, obesity, and cerebral ischemic damage. The drug shows its property in clinical trail’s patient with HIV-1 used as a traditional medicines.

International audience; The plant-derived biosurfactant aescin is naturally present in many plants and is used for treatment of disorders such as varicose veins and inflammation of veins. The hemolytic activity of this saponin is attributed to its interaction with cholesterol in the red blood cell membrane. This work investigates the phase and aggregation behavior of saponin-containing model membranes consisting of the phospholipid 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC). The aescin concentrations studied range from 1 mol% to 7 mol% with respect to the total lipid content. The methods of choice to elucidate the structural picture are small-angle scattering of X-rays (SAXS) and neutrons (SANS) and cryogenic transmission electron microscopy (cryo-TEM). SANS and SAXS revealed that at lower aescin contents vesicular structures are conserved and vesicles tend to aggregate already at aescin contents of around 1 mol%. Aggregation and vesicle deformation effects are found to be stronger when the phospholipids are in the L phase. With increasing aescin content, mixed structures, i.e. aggregated and deformed vesicles and solubilized bilayer fragments, are present. This was proven for a sample with 4 mol% aescin by cryo-TEM. An increasing aescin amount leads to membrane decomposition and free standing bilayers which tend to build stacks at high temperature. These stacks are characterized by SAXS using the modified Caillé theory. Analyses and model dependent fitting reveal formation of well-defined structures beginning at 7 mol% aescin.

Acute kidney injury (AKI) exhibited more than 1 million deaths every year worldwide; inflammation has a vital role in AKI development and considered as a risk factor in its progression. Aescin is a natural compound with anti-inflammatory vasoconstrictor and vasoprotective effects. The current study aimed to assess Aescin’s possible renal protective and therapeutic effect on potassium dichromate (PD)-induced AKI in rats. Single injection of PD was causing AKI (15 mg/kg; s.c). Rats were divided randomly into four groups. Group I: Normal control. Group II: Rats injected with PD and served as AKI group. Group III: Rats received a daily injection of Aescin for 2 weeks (3.6 mg/Kg; orally), prior PD injection and group IV after PD injection. Kidney functions, kidney contents of inflammatory and proliferative markers as toll-like receptor4 (TLR4), tumour necrosis factor-alpha (TNF-α), heat shock protein-70 (HSP-70) and insulin growth factor-1 (IGF-1) were estimated. In addition, histopathological examination was performed. Aescin improves kidney functions, alleviated inflammatory and proliferative markers and ameliorated hyaline materials in the Bowman's space and necrosis of proximal convoluted tubules that induced by PD. The present study confirmed the effectiveness of use of Aescin in AKI protection or treatment.

The saponin beta-aescin from the seed extract of the horse chestnut tree Aesculus hippocastanum has demonstrated a beneficial role in clinical therapy which is in part related to its strong interaction with biological membranes. In this context the present work investigates the self-assembly of nm-sized discoidal lipid nanoparticles composed of beta-aescin and the phospholipid 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC). The discoidal lipid nanoparticles reassemble from small discs into larger discs, ribbons and finally stacks of sheets upon heating from gel-phase to fluid phase DMPC. The morphological transition of the lipid nano-particles is mainly triggered by the phospholipid phase state change. The final morphology depends on the phospholipid-to-saponin ratio and the actual temperature. The study is conducted by small-angle X-ray scattering (SAXS) and transmission (TEM) and freeze fracture electron microscopy (FFEM) are used to cover larger length scales. Two different models, representing a disc and ribbon-like shape are applied to the SAXS data, evaluating possible geometries and molecular mixing of the nano-particles. The stacked sheets are analysed by the Caille theory.

Objective This study evaluated hepatoprotective effect of aescin (AES) and diosmin (DIO), individually or in low-dose combination in chemically induced liver injury in rats. Rats were divided into 6 groups; Group 1, control, Group 2, injected with a single dose of a mixture of corn oil and carbon tetrachloride (CCl4) to induce hepatic toxicity. Before CCl4 injection, Groups 3–6 were treated daily for 14 days with silymarin (SIL) (200 mg/kg), aescin (AES; 3.6 & 1.75 mg/kg), Diosmin (DIO; 100 & 50 mg/kg). Serum samples were analyzed for different liver function, oxidative stress and antioxidant markers. Moreover, inflammation and tissue damage were confirmed by histological staining of liver tissue sections. Results Results indicated that CCl4 elevated serum levels of all assessed liver function markers and decreased levels of key antioxidants. Administration of AES and/or DIO significantly reversed all those CCl4-induced effects. Histopathological study showed disruption of the hepatic architecture, necrosis and inflammatory cells and depositions of glycogen and protein in the tissues of CCl4-treated group. Pretreatment with DIO and/or AES significantly improved histopathological structure of liver tissue. In conclusion, low-dose combination of AES and DIO exhibited significant and preferential hepatoprotective activity compared to individual treatment with AES or DIO.

Aescin, a natural mixture of triterpene saponins, has been reported to exert anticancer effect. Recent studies show that aescin increases intracellular reactive oxygen species (ROS) levels. However, whether the increased ROS play a role in the anticancer action of aescin remains to be explored. In this study, we demonstrated that aescin (20-80 μg/mL) dose-dependently induced apoptosis and activated mammalian target of rapamycin (mTOR)-independent autophagy in human hepatocellular carcinoma HepG2 cells and colon carcinoma HCT 116 cells. The activation of autophagy favored cancer cell survival in response to aescin, as suppression of autophagy with ATG5 siRNAs or 3-methyladenine (3-MA), a selective inhibitor of autophagy, promoted aescin-induced apoptosis in vitro, and significantly enhanced the anticancer effect of aescin in vivo. Meanwhile, aescin dose-dependently elevated intracellular ROS levels and activated Ataxia-telangiectasia mutated kinase/AMP-activated protein kinase/UNC-51-like kinase-1 (ATM/AMPK/ULK1) pathway. The ROS and ATM/AMPK/ULK1 pathway were upstream modulators of the aescin-induced autophagy, as N-acetyl-L-cysteine (NAC) or ATM kinase inhibitor (KU-55933) remarkably suppressed aescin-induced autophagy and consequently promoted aescin-induced apoptosis, whereas overexpression of ATG5 partly attenuated NAC-induced enhancement in aescin-induced apoptosis. In conclusion, this study provides new insights into the roles of aescin-mediated oxidative stress and autophagy in cancer cell survival. Our results suggest that combined administration of the antioxidants or autophagic inhibitors with aescin might be a potential strategy to enhance the anticancer effect of aescin.

Medical treatment is very important in the relief of symptoms and pain related to hemorrhoidal disease, even in advanced cases with absolute surgical indication. Medical remedies containing components such as flavonoids, diosmin, calcium dobesilate, oxerutin, and horse chestnut (Aesculus hippocastanum, Hippocastanaceae family) are commonly used in the medical management of hemorrhoidal disease. The primary active constituent found in horse chestnut seed extract, aescin, is a mixture of triterpene saponins present in two forms, alpha and beta, which are distinguished by their water solubility and melting points; other constituents include bioflavonoids (quercetin and kaempferol), proanthocyanidin A2 (an antioxidant), and the coumarins fraxin and aesculin. The antiedematous, antiinflammatory and venotonic properties observed are due exclusively to aescin. Its venotonic effect was shown to be mediated by its sensitizing activity on ion channels in the vessel wall, especially to calcium, which results in an increase in contractility. It has also been proposed that enhanced release of prostaglandin F2 antagonizes the vasodilatory effects of histamine and serotonin, and venous wall damage is reduced by antagonizing proteoglycan degradation, which aids in the preservation of connective tissue integrity. Horse chestnut extract, which owes its antiseptic, venotonic, vasoprotective, and antiinflammatory properties to its aescin content, has emerged as an important agent that can facilitate the treatment of every stage of hemorrhoidal disease. In this review, we investigated these effects as well as its more recently studied apoptotic and antioxidant effects in light of experimental and clinical studies published in the literature.

This study assessed the anthelmintic activity of plant-derived compounds against gastrointestinal nematodes of goats using the egg hatch and larval motility assays. The compounds tested were saponins (digitonin and aescin) and their respective sapogenins (aglycones), hecogenin acetate and flavonoids (catechin, hesperidin, isocordoin and a mixture of isocordoin and cordoin). Additionally, cytotoxicity of active substances was analysed on Vero cell through 3-4,5-dimethylthiazol-2-yl,2,5diphenyltetrazolium bromide (MTT) and propidium iodide (PI) tests. Significant reduction on the egg hatching (P < 0.05) was seen only in the treatments with aescin (99%/EC50 = 0.67 mg mL−1) and digitonin (45%). The compounds that reduced the larval motility (P < 0.05) were digitonin (EC50 = 0.03 mg mL−1 and EC90 = 0.49 mg mL−1) and the hecogenin acetate (75%). The other sapogenins showed low anthelmintic activity. All the flavonoids showed low ovicidal (4–12%) and larvicidal (10–19%) effects. The aescin and digitonin showed low toxicity in PI test (viable cells >90%). Nevertheless, higher cytotoxicity was observed in the MTT assay, with IC50 of 0.20 mg mL−1 (aescin) and 0.0074 mg mL−1 (digitonin). Aescin and digitonin have a pronounced in vitro anthelmintic effect and the glycone portion of these saponins plays an important role in this activity.

Species of the genus Aesculus are very attractive woody ornamentals. Their organs contain numerous health-promoting phytochemicals. The most valuable of them—aescin—is used in commercial preparations for the treatment of venous insufficiency. The industrial source of aescin is horse chestnut seeds because the zygotic embryos are the main site of its accumulation. Horse chestnut somatic and zygotic embryos contain similar amount of aescin, hence somatic embryos could be exploited as an alternative source of aescin. Somatic embryogenesis, androgenesis and de novo shoot organogenesis were successfully achieved in several Aesculus species, as well as secondary somatic embryogenesis and shoot organogenesis, which enables mass production of embryos and shoots. In addition, an efficient method for cryopreservation of embryogenic tissue was established, assuring constant availability of the plant material. The developed methods are suitable for clonal propagation of elite specimens selected as the best aescin producers, the most attractive ornamentals or plants resistant to pests and diseases. These methods are also useful for molecular breeding purposes. Thus, in this review, the medicinal uses and a comprehensive survey of in vitro propagation methods established for Aesculus species, as well as the feasibility of in vitro production of aescin, are presented and discussed.

To achieve a rapid and simple detection for the active ingredients of Aescin in the extraction process using near-infrared spectroscopy (NIR) and to realize the state monitoring and quality control of the extraction process. Partial least square regression (PLS) was applied to build the near-infrared calibration models, and the applicability of the model was investigated by predicting the unknown samples in the extraction process. The correlation coefficients of the established Aescin models (A, B, C, D) were 0.9836, 0.9831, 0.9833, 0.9824, and the prediction standard deviations (SEP) were 0.05636, 0.05043, 0.02412, 0.05636, respectively. This study suggests that the proposed model has superior stability and accuracy. NIR spectroscopy technique provides a novel efficient and environmentally friendly approach to the rapid determination of four Aescin key quality indicators (A, B, C, D) in the extraction, which was solved the problem that the lack of state monitoring during the extraction of Aescin, thereby improved the quality of Aescin.

The saponin escin, extracted from horse chestnut seeds, forms adsorption layers with high viscoelasticity and low gas permeability. Upon deformation, escin adsorption layers often feature surface wrinkles with characteristic wavelength. In previous studies, we investigated the origin of this behavior and found that the substantial surface elasticity of escin layers may be related to a specific combination of short-, medium-, and long-range attractive forces, leading to tight molecular packing in the layers. In the current study, we performed atomistic molecular dynamics simulations of 441 escin molecules in a dense adsorption layer with an area per molecule of 0.49 nm2. We found that the surfactant molecules are less submerged in water and adopt a more upright position when compared to the characteristics determined in our previous simulations with much smaller molecular models. The number of neighbouring molecules and their local orientation, however, remain similar in the different-size models. To maintain their preferred mutual orientation, the escin molecules segregate into well-ordered domains and spontaneously form wrinkled layers. The same specific interactions (H-bonds, dipole–dipole attraction, and intermediate strong attraction) define the complex internal structure and the undulations of the layers. The analysis of the layer properties reveals a characteristic wrinkle wavelength related to the surface lateral dimensions, in qualitative agreement with the phenomenological description of thin elastic sheets.

Aescin nanoparticles (ANPs) were prepared using solution-enhanced dispersion by supercritical fluids (SEDS) in order to improve the dissolution properties of aescin and avoid complicated salt-forming procedures. Because aescin is a mixture, the effects of the operating conditions on the composition of the ANPs were first investigated. The compositional ratio of the four dominant active components in aescin changed with the pressure at low solution concentrations (5 mg/mL), but remained constant at higher concentrations (>10 mg/mL). Spherical ANPs with an average diameter below 50 nm was obtained at 12 MPa with solution concentration between 10 to 15 mg/mL. The morphology of ANPs was primarily influenced by the concentration and pressure but not affected by the nozzle diameter. The dissolution of aescin was increased by near 5.5 times from raw aescin to ANPs. Compared with the sodium aescinate, the ANPs exhibited the similar dissolution, but had a moderate initial dissolution burst.

Background: Aescin, a natural mixture of saponins occurring in Aesculus hippocastanum, exhibits important flebotonic properties, being used in the treatment of chronic venous insufficiency in legs. The inclusion of aescin into cyclodextrins (CDs) is a technical solution for its incorporation into the textile of stockings, but details of the physicochemistry of these host–guest systems are lacking. This work investigates the inclusion of aescin into the cavities of two native cyclodextrins, β-CD and γ-CD.Results: The continuous variation method applied to aqueous-phase 1H nuclear magnetic resonance (1H NMR) has demonstrated that the preferred CD/aescin inclusion stoichiometries are 2:1 with β-CD and 1:1 with γ-CD. The affinity constant calculated for γ-CD·aescin was 894 M−1, while for 2β-CD·aescin it was estimated to be 715 M−1. Density functional theory (DFT) calculations on the interaction of aescin Ib with CDs show that an inclusion can indeed occur and it is further demonstrated that the wider cavity of γ-CD is more adequate to accommodate this large guest. ROESY spectroscopy is consistent with the formation of a complex in which the triterpenic moiety of aescin is included into the cavity of γ-CD. The higher stability of this geometry was confirmed by DFT. Furthermore, DFT calculations were applied to determine the chemical shifts of the protons H3 and H5 of the CDs in the optimised structures of the inclusion complexes. The calculated values are very similar to the experimental data, validating the approach made in this study by NMR.Conclusion: The combination of experimental data from aqueous-state NMR measurements and theoretical calculations has demonstrated that γ-CD is the most suitable host for aescin, although the inclusion also occurs with β-CD. The geometry of the γ-CD·aescin complex is characterised by the inclusion of the triterpene segment of aescin into the host cavity.

Context: β-Aescin has anti-inflammatory, anti-oxidant and antiedematous properties. Objective: The present study investigated the hepatoprotective effect and underlying mechanisms of β-aescin in CCl4-induced liver damage. Materials and methods: Thirty-five Wistar rats were divided into six groups: normal control, CCl4 control, silymarin (50 mg/kg, p.o) and β-aescin (0.9, 1.8 and 3.6 mg/kg, i.p.) treatment for 14 d. CCl4 (1 mL/kg, i.p. for 3 d) was administered to produce hepatic damage. Ponderal changes and liver marker enzymes were estimated. Hepatic oxidative and nitrosative stress was estimated by levels of thiobarbituric acid reactive substances (TBARS), glutathione (GSH) and nitrite/nitrate. Serum TGF-β1 and TNF-α were estimated by ELISA technique. Hepatic collagen and histopathological studies were carried out. Results: β-Aescin (3.6 mg/kg) markedly decreased CCl4-induced increased levels of ALT, AST, ALP (71.77 versus 206.7, 71.39 versus 171.82, 121.20 versus 259 IU/L, respectively), total bilirubin (0.41 versus 1.35 mg/dL), TBARS (2.0 versus 8.83 nmol MDA/mg protein), nitrite/nitrate (352.50 versus 745.15 μg/mL) and increased CCl4-induced decreased GSH levels (0.095 versus 0.048 μmol/mg protein). β-Aescin (3.6 mg/kg) induced focal regenerative changes in liver and markedly decreased TBARS (2.0 versus 8.83 nmol MDA/mg protein), nitrite/nitrate (352.50 versus 745.15 μg/mL), TGF-β1 (92.28 versus 152.1 pg/mL), collagen content (110.75 versus 301.74 μmol/100 mg tissue) and TNF-α (92.82 versus 170.56 pg/mL) when compared with CCl4 control. Discussion and conclusion: The findings suggest that β-aescin has a protective effect on CCl4-induced liver injury, exhibited via its anti-inflammatory, antioxidative, antinitrosative and antifibrotic properties inducing repair regeneration of liver. Hence, it can be used as a promising hepatoprotective agent.

Objective: Aesculus indica Linn. commonly known as Indian horse chestnut belongs to family hippocastanaceae. It is extensively used in folk medicine. The whole plant is proved to possess beneficial therapeutic activities like anti-tumour, anti-diabetic and anti-oxidant etc. However no approved marketed formulation of any of its active ingredients is available in any country of the world till date. There is also a subtle mention of its toxicity in some studies. Its uses can be exploited commercially in a better way if its active pharmaceutical ingredients are able to be separated from its toxic ingredients. Succinctly, the plant can be used commercially in future only when its toxicity is managed somehow. The review briefly summarizes its beneficial effects and the problem associated with it as a counterpart with a brief mention of on-going medicinal research on it. Data Source: The data is collected from different databases i.e. Science Direct, PubMed, Google and Google Scholar. Review Methods: Bibliographic investigation was carried out by analysing various research and review papers. Results and Conclusion: Aesculus indica can be used to develop variety of products with high nutritional and medicinal values. The compound Aescin (a triterpenoids saponin) has been isolated from this plant and i.e. used to prevent several ailments such as rheumatism, skin diseases, relieve headache and wound healing. It also contains some phytoconstituents which have high potency to be used in cosmetics.

Worldwide, oral squamous cell carcinoma (OSCC) is the sixth most common cancer & ranked as second most common cancer in India. The most implicated etiologic factors in the development of oral cancer is the use of tobacco, alcohol & also HPV infection. Aescin, a pentacyclic triterpenoid saponin mixtures derived from Aesculus hippocastanum L (horse chestnut) that has been used as traditional herbal medicine, exists the pharmacodynamic properties, anti- edematous, anti-inflammatory & antitumor. The present study, Tumors were induced by applying 0.5% DMBA topically to hamster buccal pouch (HBP) thrice a week for 14 weeks & Aescin was orally administered at a various dose of 40 mg/kg body weight on alternate days of DMBA painting. We observed, reduced the tumor incidence, diminished lipid peroxidation, & elevation of antioxidants, liver marker enzymes by Aescin. Furthermore, aescin induced apoptosis in DMBA induced tumor bearing animals significantly inhibited cell to cell communication of Notch-1, Notch-2 & its ligand & Jagged-1 leads to inhibited expression of the Notch downstream signaling target Hes-1 & Hey-1 analyzed by immunohistochemical analysis. These results clearly suggest that Aescin prevents lipid peroxidation, protects the antioxidant defense system, prohibit the dysplasia & anti-carcinogenic potential. we concluded; the results provide an important new insight of Notch signaling pathway & aescin might be developed as a potential chemo preventive agent for oral cancer. Key words: Oral cancer, aescin, antioxidant, apoptosis, chemopreventive, Notch signaling Citation Format: Hemavardhini Ramadoss, Suresh Kathiresan, Sanjay Gupta. Apoptosis & anti-proliferative nature of Aescin on 7,12 dimethylbenz[a]anthracene (DMBA) induced hamster buccal pouch carcinogenesis via downregulating Notch signaling [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 129.

The incidence of Head and Neck cancer in increasing worldwide. The major risk factor for these cancers includes viral infection, smoking, alcohol and tobacco consumption. Natural agents have often exhibited medicinal properties with usefulness in treating various human ailments including cancer. Aescin is one of the most prominent constituent of Aesculus hippocastanum L. (Hippocastanaceae) seeds has been conventionally used as medicinal herb. We determined the anticancer effect of aescin on human oral carcinoma (KB) cells and human laryngeal carcinoma (HEp-2) cells. Treatment of KB cells with 25µM and HEp-2 cells with 22µM aescin for 24 hours suppressed proliferation in these cells and induced morphological changes consistent with apoptosis. Exposure of both cell lines to aescin resulted in marked increase in reactive oxygen species, loss of cell viability, and reduction of cellular glutathione levels resulting in depolarization of mitochondrial membrane potential thereby increasing oxidative DNA damage in KB and HEp-2 cells. Furthermore, aescin-mediated apoptosis in these cells progress via notch signaling as shown by downregulation of Notch1, Notch2, Jagged-1, HES1, and HEY1 expression. Taken together, our results demonstrate that aescin initiate apoptosis in KB and HEp-2 cells by increasing oxidative damage implying that aescin might be developed as a potential chemotherapeutic agent for the treatment of human oral and laryngeal cancer. Key word: Oral carcinoma, Aescin, apoptosis, anti-proliferative, Notch signaling Citation Format: Suresh Kathiresan, Hemavardhini Ramadoss, Sanjay Gupta. Aescin, a triterpene saponin induces apoptosis & oxidative damage in human oral & laryngeal cancer cells via notch signaling [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 6557.

Horse chestnut (Aesculus hippocastanum L.) is a deciduous tree which is widely used on urban green spaces in Serbia. Also, the seed of horse chestnut is an important raw material in pharmaceutical industry, owing to its medicinal properties. In this study, the content of fatty acids, flavonoids, aescin and heavy metals in seeds of 11 horse chestnut genotypes growing in urban conditions was investigated. A total of 16 fatty acids were identified. Unsaturated fatty acids in seed samples were the most abundat. Among the saturated fatty acids, palmitic acid was the dominant one, with a total average content of 8.12%. Oleic acid, with the total average content of 53.46%, was the most abundant unsaturated fatty acid. The results of this study indicated the presence of the following flavonoids: quercetin, kaempferol and rutin. Rutin was the most abundant, averagely contributing with 6.071 μg/g to the total quantified polyphenolics in all samples. The total aescin content ranged from 0.82% to 4.16%. Fruits of park and street trees from small- and middle-sized cities can be used in pharmaceutical industry as raw material. Differences between different urban site categories and localities were detected, which indicates the biochemical plasticity of the seeds to urban morphology. Supplementary material: [http://cherry.chem.bg.ac.rs/handle/123456789/3807]

In the present work, we study the interaction of the saponin aescin with the nonsteroidal anti-inflammatory drug (NSAID) ibuprofen at concentrations of 1.2-2.5 mM. These amounts are higher than those usually used for medication (10-300 mu M) to show possible structures and formulations for orally absorbed drug delivery systems. It is shown how the interaction of both substances, separately or together, alters the thermotropic phase behavior of the 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC) bilayer in the presence of different amounts of aescin, ranging from 20 mu M to 1 mM. The methods of choice are differential scanning calorimetry (DSC), and additionally wide-angle (WAXS) and small angle X-ray scattering (SAXS). We found that these two additives, aescin and ibuprofen, alter the temperature-dependent structural appearance of the DMPC membrane depending on the aescin and drug content. The presence of the saponin and the drug become visible on different length scales, i.e., ranging from a global structural change to inner-membrane interactions. DSC reveals that the drug and saponin alter the cooperativity of the DMPC phase transition in a concentration-dependent manner. Furthermore, there is a significant difference between the drug-containing compared to the drug-free systems. By WAXS, we could resolve that aescin reverses the strong impact of ibuprofen on the diffraction peak of DMPC. Both molecules interact strongly with the phospholipid headgroups. This becomes visible in a changing area per lipid and shifting phase transition to higher temperatures. SAXS experiments reveal that the addition of ibuprofen leads to major morphological changes in the phospholipid bilayer. SAXS experiments performed on representative samples do not only cover the drug saponin interaction within the bilayer from the structural perspective but also confirm the visually observed macroscopic concentration and temperature-dependent phase behavior. Vesicular shape of extruded samples is conserved at low aescin contents. At intermediate aescin content, aggregation between vesicles occurs, whereby the strength of aggregation is reduced by ibuprofen. At high aescin contents, DMPC bilayers are solubilized. The kind of formed structures depends on temperature and drug content. At low temperature, separated bilayer sheets are formed. Their size increases with ibuprofen in a concentration dependent manner. At high temperature, the drug-free system reorganizes into stacked sheets. Whereas sheets at 5 mol % ibuprofen close to vesicles, the ones with 10 mol % of the drug increase massively in size. Altogether, ibuprofen was found to rather enhance than inhibit structural and thermotropic membrane modifications induced by the aescin on the DMPC model membrane.

This study is aimed to evaluate the potential effects of sodium aescinate (SA, the sodium salt of aescin) on wound healing in streptozotocin-induced diabetic rats. An excision skin wound was created in diabetic rats, and the wounded rats were divided into three groups: I) control group, II) gel-treated group, and III) SA-treated group. The control group wounds received topically normal saline once daily for 19 days. The gel-treated and SA-treated wounds received topically 400 μl of pluronic F-127 gel (25%) and 400 μl of SA (0.3%) in pluronic gel, respectively, once daily for 19 days. SA application in diabetic rats increased the wound contraction and significantly decreased the level of the inflammatory cytokine tumor necrosis factor-alpha (TNF-α) in comparison to the gel-treated group and control group. SA application in diabetic rats also resulted in a marked increase in the level of anti-inflammatory cytokine interleukin-10 (IL-10) and activities of antioxidant enzymes superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GSH-Px) compared to the other groups. Histopathologically, SA-treated wounds showed better granulation tissue dominated by marked fibroblast proliferation, and wounds were covered by thick regenerated epithelial layer. Additionally, the application of only pluronic gel produced some beneficial effects in some parameters in comparison to control group, but most of them were not significantly different. These findings demonstrated that SA may effectively control and improve wound healing in diabetic rats via its anti-inflammatory and antioxidant activities.

Vesicle shape and bilayer parameters are studied by small-angle X-ray (SAXS) and small-angle neutron (SANS) scattering in the presence of the saponin aescin. We confirm successful incorporation of aescin molecules by analysis of the radii of gyration RG and study furthermore the impact of aescin incorporation on bilayer thickness parameters from the neutron and X-ray perspective. Additionally, the bending elasticity (κ) of these 1,2-dimyristoyl-sn-glycero-3-phosphocholine vesicle bilayers is studied in the presence of aescin. Neutron spin-echo spectroscopy (NSE) allows to detect subtle changes in the dynamics and κ of lipid membranes. Changes of κ are detectable at temperatures below and above the main phase transition temperature Tm of the lipid. The impact of aescin is much more significant below Tm. It has been found that below Tm the addition of aescin to the vesicles decreases the value of κ and softens the bilayer. Above Tm the value of κ increases with increasing aescin content and the bilayer becomes more rigid. Altogether, we demonstrate by analysis of the structure and dynamics of the vesicles that the impact of aescin strongly depends on the lipid state. Below Tm the membrane becomes fluidized and softer, above Tm solidified and stiffer compared to a DMPC membrane without additive at similar conditions.

Rationale: Pachydermoperiostosis (PDP) is a rare hereditary disorder that affects the skin and bones. PDP is characterized by periostosis, digital clubbing, and pachydermia. Previous studies demonstrated that increased prostaglandin E2 (PGE2) levels resulting from defective protein degradation pathways play a crucial role in PDP pathogenesis, and males were more commonly and severely affected than females. Moreover, nearly all PDP patients suffer from refractory arthralgia. Although several different treatment modalities are used for PDP, therapy for this disease remains challenging. Patients concerns: Two cases of PDP showing symptoms consistent with polyarthritis and arthralgia that mainly affected the knees and ankles. Diagnoses: The diagnostic criteria for PDP include digital clubbing, periostosis, and pachydermia. The 2 patients were diagnosed as PDP based on the finger clubbing, facial cutis furrowing, knee and ankle arthritis, and radiographic evidence of periosteal proliferation. Interventions: Patient 1 had massive joint effusion that was treated by oral administration of etoricoxib and aescin combined with arthroscopic synovectomy, whereas Patient 2 had mild joint swelling and accepted only oral medication. Outcomes: Clinical symptoms of the 2 patients greatly improved after the treatment. During the 1-year follow-up, the patient experienced no adverse effects or recurrence. Lessons: The therapeutic results showed that oral etoricoxib could reduce inflammation and retard progression of pachydermia, or even relieve facial skin furrowing, but had limited efficacy for arthralgia. However, oral aescin had satisfactory efficacy for arthralgia. Thus, etoricoxib combined with aescin is a safe and effective treatment for PDP. Meanwhile, arthroscopic synovectomy can be used to treat joint effusion, but had no therapeutic effect on arthralgia. Therefore, postoperative oral medications would be needed as subsequent therapy for joint problems. In conclusion, this study proposes an effective and safe treatment plan to address symptoms experienced by PDP patients.

This work describes the functionalization of a natural and a man-made textile substrate, namely cotton and polyamide 6.6, with inclusion agents, β-cyclodextrins (β-CDs) that are able to release gradually to the user active ingredients. In this study we used aescin (aesculus hippocastanum extract), which is a natural agent with benefits for the treatment of varicose veins. 1H NMR and UV–vis data supported the role of β-CDs as an aescin complexing agent, and the covalent nature of the linkage between β-cyclodextrins and the textile substrates, which showed a wash fastness to more than 45 washing cycles.

The saponin aescin from the horse chestnut tree is a natural surfactant well-known to self-assemble as oriented-aggregates at fluid interfaces. Using model membranes in the form of lipid vesicles and Langmuir monolayers, we study the mixing properties of aescin with the phase-segregating phospholipid 1,2-dimyristoyl-sn-glycero-phosphocholine (DMPC). The binary membranes are experimentally studied on different length scales ranging from the lipid headgroup area to the macroscopic scale using small-angle X-ray scattering (SAXS), photon correlation spectroscopy (PCS), and differential scanning calorimetry (DSC) with binary bilayer vesicles and Langmuir tensiometry (LT) with lipid monolayers spread on the surface of aescin solutions. The binary interaction was found to strongly depend on aescin concentration in two well differentiated concentration regimes. Below 7 mol %, the results reveal phase segregation of nanometer-sized aescin-rich domains in an aescin-poor continuous bilayer. Above this concentration, aescin-aescin interactions dominate, which inhibit vesicle formation but lead to the formation of new membrane aggregates of smaller sizes. From LT studies in monolayers, the interaction of aescin with DMPC was shown to be stronger in the condensed phase than in the liquid expanded phase. Furthermore, a destructuring role was revealed for aescin on phospholipid membranes, similar to the fluidizing effect of cholesterol and nonsteroidal anti-inflammatory drugs (NSAIDs) on lipid bilayers.

Introduction. Primrose or primula ( Primula L.) is a genus of Primulaceae family plants, which includes about 500 species, distributed throughout the globe. 67 species (according to some authors – 80) grow in the CIS countries, in Ukraine – nine of them; the most common is Primula veris . Primula veris L. is a medicinal, vitamin, nutritional, honeyed, dyeing and decorative plant. Its chemical composition is insufficiently studied despite the widespread usage of primula in medical practice. There is no information about research of saponins contents in leaves and underground organs of Primula veris of Ukrainian flora in the available sources of scientific literature. The aim of the study – to determine the content of saponins in leaves and rhizomes with roots of Primula veris . Research methods. Water and spirituous-water extracts of leaves and rhizomes with roots of Primula veris were studied for saponins content. The aqueous extract of primula was used for tests on foam formation and some sedimentary reactions. The determination of chemical nature of saponins was also done. Quantitative content of saponins was determined by spectrophotometric method on a spectrophotometer Lambda 25 UV at a wave length 381 nm with following conversion to aescin. Results and discussion. The emergence of stable foam and positive reactions with 10 % solution of basic lead acetate and 1 % alcoholic solution of cholesterol showed the presence of saponins in the leaves and rhizomes with roots of of the studied plant. It was found out during determining the chemical nature of saponins that the leaves and rhizomes with roots of Primula veris contain triterpene saponins – (1.27 ± 0.02) % and (3.69 ± 0.02) % respectively in conversion to aescin. Conclusion. It was experimentally proved that the leaves and rhizomes with roots of Primula veris contain triterpene saponins. For the first time it was defined quantitative content of saponins in leaves and underground organs of Primula veris from Ternopil region. The content of saponins in leaves was 1.27 %, in rhizomes with roots – 3.69 % in conversion to escin.

Objective To verify the trypanocidal effectiveness of aescin and aescin liposomes against Trypanosoma evansi in vitro and in vivo . Methods Aescin and aescin liposomes were used in vitro on trypomastigotes at different concentrations (0.5%, 1.0% and 2.0%) and exposure times (0, 1, 3, 6 and 9 h). In vivo tests were performed using mice as the experimental model. Trypanosome evansi infected mice were treated with aescin and aescin liposomes with doses of 60 and 100 mg/kg during 4 d. Results The three concentrations tested in free form and nanoencapsulated showed trypanocidal activity in vitro , completely eliminating the parasites in small concentration after 6 h of assay. Animals treated with aescin (100 mg/kg) and aescin liposomes (100 mg/kg) showed increase in longevity, however without curative effect. Conclusions Active compounds present in natural products, such as aescin, may potentiate the treatment of trypanosomosis when used in association with other trypanocidal drugs.

Introduction Escin is the predominant active constituent of Aesculus hippocastanum seed extract. In addition, Aesculus hippocastanum seed contains flavonols, glycosides (Aesculin, esculin) and triterpenoid saponins (aescin, escin). The antioxidant biological effect of escin means ability to protect against free radicals and toxins production in lung cells. Also, several in vivo studies have shown that low doses of escin are anti-tumour and anti-inflammatory in different cell lines. In this study was investigated potential cytotoxic effects of escin-loaded solid lipid nanoparticles on the morphology of A549 cell line. Material and methods The MTT method was used to determine the cytotoxic effects of the nano-formulation in A549 cells. Firstly, the nano formulation of escin was prepared and the A549 cells were exposed to this compound at different concentrations for 24 hours, then read on the ELISA reader at 570 nm wavelength. Based on these results, IC 50 value was found. Morphological changes on the A549 cells caused by escin nano-formulation were examined under confocal microscope. For this manner A549 cells were incubated with the IC 50 concentration of nano-formulation for 24 hours and cells were double stained with phalloidin and acridine orange. Changes on the cell morphology were photographed under a confocal microscope. Results and discussions The viability of the treated cells decreased with the increase of the applied concentration. IC 50 value of 3.20 µM for 24 hours. On the confocal micrographs of A549 cells exposed to IC 50 value of escin nano-formulation for 24 hours was seen many morphological alterations as disintegrated and deformed nuclei, chromatin and cytoskeleton, chromatin condensation also cell shrinkage and holes on cytoskeleton. Conclusion According to our laboratory studies and results, escin nano-formulation has been shown that escin-loaded solid lipid nanoparticles induced apoptosis in human lung adenocarcinoma (A549) cells and caused morphological changes on these cells. As escin nano-formulation doses increased the viability of the treated A549 cells decreased. Escin nano-formulation caused holes on the skeleton of these cells and caused cells to shrink. It also triggered apoptosis of these cells. We suggest this drug as an alternative agent in the treatment of cancer but further investigations are needed.

Aesculus hippocastanum L. (Sapindaceae), vulgarly known as horse chestnut, is a native European plant, but widely distributed all over the world. The parts used in medicine are the seeds and the bark of young branches. The seed extract has been marketed as a phytotherapeutic product and it is extensively recommended for treatment of chronic venous insufficiency, traumatic edema, hemorrhoid and postoperative edema. Most of the beneficial effects of horse chestnut are attributed to its principal component aescin. The present study aims to evaluate the citotoxic, genotoxic, mutagenic, and antioxidant activity of this seed extract by performing a set of bacterial and cell-free tests. Phytochemical screening and thin layer chromatography confirmed the presence of aescin as well as others chemical constituents in the studied extract. Negative results were obtained for the occurrence of double-strand breaks in pBCKS plasmid DNA. Two different effects were detected in Escherichia coli strains CC104 and CC104mutMmutY: mutagenic (non-oxidative lesions) and antioxidant. The mutagenic effect was confirmed by Kado assay, where the extract was considered weakly mutagenic towards frameshift mutations. Moreover, the extract demonstrated to protect bacterial cells from lesions induced by mitomycin C, suggesting an antioxidative potential. A. hippocastanum seed extract demonstrated to be genotoxic and weakly mutagenic, as well as antioxidative.

Aescin has many physiological functions that are highly relevant to spinal cord injury (SCI), including anti-inflammation, anti-oxidation, anti-oedema, and enhancing vascular tone. The present study investigated the putative therapeutic value of aescin in SCI, with a focus on its neuroprotective, anti-inflammatory, and anti-oxidative properties. Sodium aescinate (1.0 mg/kg body weight) or equivalent volume of saline was administered 30 min after injury by intravenous injection, with an additional dose daily for seven consecutive days after moderate SCI in rats. After contusion injury of the 8th thoracic (T8) spinal cord, aescin-treated rats developed less severe hind limb weakness than saline controls, as assayed by the Basso-Beattie-Bresnahan scale, the beam walking test, and a footprint analysis. The improved locomotor outcomes in aescin-treated rats corresponded to markedly decreased immune response, oxidative stress, neuronal loss, axon demyelination, spinal cord swelling, and cell apoptosis, measured around T8 after impact. Our data suggest aescin treatment as a novel, early, neuroprotective approach in SCI. Given the known safety of aescin in clinical applications, the results of this study suggest that it is a good candidate for SCI treatment in humans.

Methyl parathion, a highly cytotoxic insecticide, has been used in agricultural pest control for several years. The present study investigated the protective effect of sodium aescinate (SA, the sodium salt of aescin) against liver injury induced by methyl parathion. Forty male Sprague-Dawley rats were randomly divided into 5 groups of 8 animals: the control group; the methyl parathion (15 mg/kg) poisoning (MP) group; and the MP plus SA at doses of 0.45, 0.9 and 1.8 mg/kg groups. Alanine aminotransferase (ALT), aspartate aminotransferase (AST) and acetylcholinesterase (AChE) in the plasma were assayed. Nitric oxide (NO) and antioxidative parameters were measured. Histopathological examination of the liver was also performed. The results revealed that SA had no effect on AChE. Treatment with SA decreased the activities of ALT and AST, and the levels of malondialdehyde and NO. Treatment with SA also increased the level of glutathione and the activities of superoxide dismutase and glutathione peroxidase. SA administration also ameliorated liver injury induced by methyl parathion poisoning. The findings indicate that SA protects against liver injury induced by methyl parathion and that the mechanism of action is related to the antioxidative and anti-inflammatory effects of SA.

To investigate the effect and underlying mechanism of aescin on acute liver injury induced by endotoxin, liver injury was established by injecting lipopolysaccharide (LPS) in mice. Animals were assigned to seven groups: the control group and groups treated with LPS (40 mg/kg), aescin (3.6 mg/kg), LPS plus dexamethasone (4 mg/kg) and LPS plus aescin (0.9, 1.8 or 3.6 mg/kg). Hepatic histopathological changes were examined under a light microscope. Activities of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in serum were determined. Levels of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), nitric oxide (NO) and antioxidative parameters in liver homogenate were measured. Glucocorticoid receptor (GR), 11 beta-hydroxysteroid dehydrogenase type 1 (11β-HSD1) and 11 beta-hydroxysteroid dehydrogenase type 2 (11β-HSD2) expressions in liver were determined by western blotting. Treatment with escin could inhibit immigration of inflammatory cells, alleviate the degree of necrosis, and decrease serum ALT and AST activities. Aescin also down-regulated levels of inflammation mediators (TNF-α, IL-1β and NO) and 11β-HSD2 expression in liver, up-regulated GR expression, enhanced endogenous antioxidative capacity, but have no obvious effect on 11β-HSD1 expression in liver. The findings suggest aescin has protective effects on endotoxin-induced liver injury, and the underlying mechanisms were associated with its anti-inflammatory effects, up-regulating GR expression, down-regulating 11β-HSD2 experssion, and antixoidation.

In phytomedicine complex extracts consisting of phenolics, monoterpenes or saponins are traditionally used. It is often impossible to attribute the biological activity of an extract to one or few compounds. As an explanation of the superior activity of extracts, a synergistic effect of combinations of active compounds has been suggested. Since lipophilic monoterpenes or saponins targeting the biomembrane usually accompany polar polyphenols in phytomedical preparations, we decided to investigate their effect as single substances and in combination to gain further insight into potential synergistic effects of herbal medicine. Combinations of the monoterpenes α-pinene, thymol and menthol with the monodesmosidic saponins digitonin, aescin, glycyrrhizic acid and Quillaja saponin demonstrated strong synergistic activity. The IC50 of haemolysis was lowered by a factor of 10–100 from 316 μg/ml to 2 μg/ml for aescin, 157 μg/ml to 11 μg/ml for Quillaja saponins and 20 μg/ml to 3 μg/ml for digitonin when combined with thymol. A similar significant synergistic cytotoxicity occurred both in HeLa and Cos7 cells by combining the α-pinene, thymol and menthol with the saponins. The IC50 of glycyrrhizic acid was lowered by a factor 100 from around 300 μg/ml to around 1–10 μg/ml and the IC50 of aescin, digitonin and Quillaja saponins about the factor 10. Monoterpenes and monodesmosidic saponins have a common target, the biomembrane, which is present in all animal, fungal and bacterial cells. Disturbance of membrane fluidity and permeability is the mode of action. This activity is non-specific which makes it extremely difficult for bacteria and fungi to develop resistance. This explains the overall success of these molecules as defence chemicals in the plant kingdom. The synergistic effect of combinations of saponins with monoterpenes opens a complete new field of possible applications in medicine to overcome resistance in multidrug resistant microbial and human cell.

BACKGROUND: Saponins are a class of secondary plant metabolites consisting of a sugar moiety glycosidically linked to a hydrophobic aglycone (sapogenin) that often possess insecticidal activities. Four saponins were selected: two triterpene saponins,Q.saponaria saponins and aescin, and two steroidal saponins, digitonin and diosgenin. Their effects were investigated on an important pest species and a model piercing-sucking insect, the pea aphidAcyrthosiphonpisum. The triterpeneQ.saponaria saponins bark saponin received special attention because of its high activity. Aphids were challenged by oral and contact exposure to demonstrate aphicidal activities, and in choice experiments to support use as a natural deterrent. RESULTS: When aphids were exposed to supplemented artificial diet for 3 days, a strong aphicidal activity was recorded for three of the four saponins, with an LC50 of 0.55 mg mL −1 for Q. saponaria saponins, 0.62 mg mL −1 for aescin and 0.45 mg mL −1 for digitonin. The LT50 values ranged between 1 and 4 days, depending on the dose. For diosgenin, only low toxicity (14%) was scored for concentrations up to 5 mg mL −1 . In choice experiments with treated diet, a deterrence index of 0.97 was scored for Q. saponaria saponins at 1 mg mL −1 . In contrast, direct contact showed no repellent effect. Spraying of faba bean plants with Q.saponaria saponins resulted in an LC50 of 8.2 mg mL −1 . Finally, histological analysis in aphids fed withQ.saponaria saponins demonstrated strong aberrations of the aphid gut epithelium, and exposure of midgut CF-203 cell lines toQ.saponaria saponins invitro confirmed the cytotoxic effect. CONCLUSIONS: The present insect experiments provide strong evidence that saponins, as tested here with triterpene Q.saponaria saponins, can be useful as natural aphicides and deterrents. Furthermore, the insect midgut epithelium is suggested to be a primary target of saponin activity. c � 2011 Society of Chemical Industry

The colorimetric assay used for saponin quantification in plant extracts is subject to interference by common solvents used to extract the saponins from plant materials. Therefore, the degree of interference of ten common solvents was investigated. It was found that the presence of acetone, methanol and n-butanol in the reaction solution caused an intense darkening of the reaction solution in the absence of saponins, which likely could lead to erroneous saponin content values. Using aescin to construct standard curves with different solvents&mdash, such as water, ethanol, and methanol&mdash, also showed significant differences in the standard curves obtained, which led to different values when they were applied to quantify the saponin content of an ethanol extract from dried and powdered Gac (Momordica cochinchinensis Spreng) seed kernels. To improve the method, a solvent evaporation step was added prior to the colorisation reaction to prevent undesired solvent interference during the reaction step. Using this modified protocol for the aescin standard curve and the Gac seed kernel extract eliminated any solvent interference. Thus, this improved protocol is recommended for the quantification of the saponin content of plant extracts irrespective of which extraction solvent is used.

Background: Gac (Momordica cochinchinensis Spreng.) seeds contain saponins that are reportedly medicinal. It was hypothesised that the extraction of saponins from powdered Gac seed kernels could be optimised using microwave-assisted extraction (MAE) with ethanol as the extraction solvent. The aim was to determine an appropriate ethanol concentration, ratio of solvent to seed powder and microwave power and time for extraction. Whether or not defatting the Gac seed powder had an impact on the extraction of saponins, was also determined. Methods: Ethanol concentrations ranged from 60–100% were used to compare total saponins content (TSC) extracted from full-fat and defatted Gac seeds. Ratios of solvent to Gac seeds ranged from 10 to 100 mL g−1 and microwave conditions ranged from 1–4 cycles at power levels ranged from 360–720 W, were examined successively to evaluate their efficiency in extracting saponins from full-fat Gac seeds. Results: A four-fold higher of TSC was obtained in extracts from full-fat Gac seed powder than from defatted powder (100 vs. 26 mg aescin equivalents (AE) per gram of Gac seeds). The optimal parameters for the extraction of saponins were a ratio of 30 mL of 100% absolute ethanol per g of full-fat Gac seed powder with the microwave set at 360Wfor three irradiation cycles of 10 s power ON and 15 s power OFF per cycle. Conclusions: Gac seed saponins could be efficiently extracted using MAE. Full-fat powder of the seed kernels is recommended to be used for a better yield of saponins. The optimised MAE conditions are recommended for the extraction of enriched saponins from Gac seeds for potential application in the nutraceutical and pharmaceutical industries.

The effect of horse-chestnut seed extract (standardized on aescin; Venostasin retard) was assessed in a randomized placebo-controlled crossover double-blind trial of 22 patients with proven chronic venous insufficiency by measuring the capillary filtration coefficient and the intravascular volume of the lower leg by venous-occlusion plethysmography. Three hours after taking two capsules of Venostasin (600 mg; each capsule containing 50 mg aescin) the capillary filtration coefficient had decreased by 22%, whereas after administration of an identical-looking placebo capsule it rose but slightly over three hours. The difference in the effect of Venostasin and placebo is statistically significant (P = 0.006). The intravascular volume was reduced 5% more after Venostasin than the placebo, but this is not statistically significant. It is concluded that Venostasin has an inhibitory effect on oedema formation via a decrease in transcapillary filtration and thus improves oedema-related symptoms in venous diseases of the legs.

Human vascular endothelial cells (HUVECs) were exposed to CoCl2 as an in vitro model of hypoxia. Expression of VCAM-1 (vascular cell adhesion molecule), reduction of PECAM-1 (platelet endothelial cell adhesion molecule) and cytoskeletal changes without alterations in cell viability were observed. HUVECs were also exposed to Escherichia coli lipopolysaccaride (LPS) as an in vitro model of inflammation: significant IL-6 release was measured. Pre-treatment of HUVECs with aescin prevented, in a concentration-dependent fashion (0.1-1 microM), the action of CoCl2 on VCAM-1 and PECAM-1, also preserving endothelial cell morphology. Furthermore, aescin pre-treatment reduced IL-6 release from LPS-activated vascular endothelium.

Aescin (escin) derived from the seeds of horse chestnut (Aesculus hippocastanum L.) is a natural mixture of triterpene saponins exhibiting a wide variety of pharmacological properties, including antiinflammatory, analgesic, and antipyretic activities. However, data concerning antifungal activities of these compounds are limited. This study aims to evaluate the in vitro antifungal susceptibility of Candida glabrata clinical isolates to α-aescin sodium, β-aescin crystalline and β-aescin sodium using the disk diffusion (DD) and broth microdilution (BMD) methods. Moreover, the influence of subinhibitory concentration (0.5×MIC) of β-aescins on the nystatin MIC was also studied. In general, the results obtained by the DD assay correlated well with those obtained by the BMD method. Both β-aescins effectively inhibited the growth of all 24 strains tested. The minimum inhibitory concentration (MIC) values ranging from 8 to 32 μg/ml for β-aescin crystalline, whereas those of β-aescin sodium were slightly lower and ranged from 4 to 16 μg/ml. In contrast, α-aescin sodium was found to be completely ineffective against the strains studied. MIC values of nystatin were reduced 2-16-fold and 2-4-fold in the presence of subinhibitory concentration of β-aescin crystalline and β-aescin sodium, respectively. Results of the present study may suggest the additive interaction between β-aescin and nystatin.

Extracts of Aesculus hippocastanum (horse chestnut) seed have been used in the treatment of chronic venous insufficiency, edema, and hemorrhoids. Most of the beneficial effects of horse chestnut are attributed to its principal component β-escin or aescin. Recent studies suggest that β-escin may possess anti-inflammatory, anti-hyaluronidase, and anti-histamine properties. We have evaluated the chemopreventive efficacy of dietary β-escin on azoxymethane-induced colonic aberrant crypt foci (ACF). In addition, we analyzed the cell growth inhibitory effects and the induction of apoptosis in HT-29 human colon cancer cell line. To evaluate the inhibitory properties of β-escin on colonic ACF, 7-week-old male F344 rats were fed experimental diets containing 0%, 0.025%, or 0.05% β-escin. After 1 week, the rats received s.c. injections of azoxymethane (15 mg/kg body weight, once weekly for 2 weeks) or an equal volume of normal saline (vehicle). Rats were continued on respective experimental diets and sacrificed 8 weeks after the azoxymethane treatment. Colons were evaluated histopathologically for ACF. Administration of dietary 0.025% and 0.05% β-escin significantly suppressed total colonic ACF formation up to ∼40% (P < 0.001) and ∼50% (P < 0.0001), respectively, when compared with control diet group. Importantly, rats fed β-escin showed dose-dependent inhibition (∼49% to 65%, P < 0.0001) of foci containing four or more aberrant crypts. To understand the growth inhibitory effects, HT-29 human colon carcinoma cell lines were treated with various concentrations of β-escin and analyzed by flow cytometry for apoptosis and cell cycle progression. β-Escin treatment in HT-29 cells induced growth arrest at the G1-S phase, which was associated with the induction of the cyclin-dependent kinase inhibitor p21WAF1/CIP1, and this correlated with reduced phosphorylation of retinoblastoma protein. Results also indicate that β-escin inhibited growth of colon cancer cells with either wild-type or mutant p53. This novel feature of β-escin, a triterpene saponin, may be a useful candidate agent for colon cancer chemoprevention and treatment. [Mol Cancer Ther 2006;5(6):1459–66]

A thin-layer chromatographic (TLC) method is developed to analyze the total saponin content, also referred to as the aescin content, in a herbal medicinal product (HMP) containing two dry extracts in capsules. The capsules contain 250 mg of Aesculus hippocastanum dry extract, 120 mg of Vitis vinifera dry extract and 50 mg of excipients. After a purification step using C18 solid phase extraction (SPE) cartridges, the samples are analyzed on a silica-gel HPTLC plate with the upper layer of a mixture of acetic acid/water/butanol (10/40/50 v/v/v) as the mobile phase. Spots are visualized by spraying with anisaldehyde reagent and heating the plate for 5–10 min (100–105 °C) and measured at a wavelength of 535 nm. This method, applicable for the quality control and stability investigation of both the Aesculus dry extract and HMP capsules thereof containing Vitis dry extract in combination with the Aesculus dry extract, is validated according to the International Conference on Harmonization (ICH) guidelines. The proposed assay method is specific for aescin in the presence of Vitis dry extract and formulation excipients. Analysis of stressed samples in forced degradation tests proves the method to be applicable for stability evaluation. The standard aescin curve is linear (r > 0.99) over a concentration range of 0.16–0.80 μg/spot. Recovery from the HMP capsules is statistically equal to 100%. The precision of the method with respect to time and concentration is acceptable, with relative standard deviation (RSD) values of 1.28 and 1.49%, respectively.

PURPOSE: To evaluate the efficacy of the addition of tamsulosin to our standard expulsive pharmacologic therapy for the treatment of distal-ureteral stones. PATIENTS AND METHODS: A series of 96 patients referred to our department for the management of symptomatic distal-ureteral calculi were randomly divided into group 1 (N = 46) who received diclofenac (100 mg/daily) plus aescin (80 mg/daily) and group 2 (N = 50) who received the same therapy plus tamsulosin (0.4 mg/daily) for a maximum of 2 weeks. There were no differences between the groups with respect to age, sex, or stone size. The primary endpoint was the expulsion rate. Expulsion time, need for analgesics, need for hospitalization, and drug side effects were the secondary endpoints. RESULTS: The expulsion rate was significantly higher in group 2 (90%) than in group 1 (58.7%; P = 0.01), and group 2 achieved stone passage in a shorter time (mean 4.4 v 7.5 days, respectively; P = 0.005). Lower analgesic use was found in group 2 (P = 0.003), as well as significantly fewer hospitalizations for recurrent colic (P = 0.01). Both groups experienced few side effects associated with expulsive therapy. CONCLUSIONS: A conservative approach should be considered as an option in the management of uncomplicated distal-ureteral stones. Even if the best pharmacologic expulsive regimen remains to be established, the use of the selective alpha-blocker tamsulosin is recommended in this setting.

This study reports experimental measurements on the ability of aqueous biosurfactant solutions (aescin, lecithin, rhamnolipid, saponin, and tannin) at removing Ekofisk crude oil from a laboratory contaminated soil under varying washing conditions. The oil removal performance of the biosurfactants was evaluated against a synthetic anionic surfactant (sodium dodecyl sulphate, SDS) using distilled water as a base case. The washing parameters and ranges tested were temperature, time, shaking speed, volume/mass ratio, and surfactant concentrations. Results indicated that washing temperature was the most influential parameter on the oil removal whilst washing time was the least. It was possible to obtain more than 80% oil removal at 50 °C for all the surfactant solutions, except lecithin, which yielded less than 15% removal. However, saponin, lecithin, aescin, and tannin removed less than 50% crude oil when tested at a temperature of 20 °C and other parameters. Soil washing was found to have considerable potential in removing crude oil from the contaminated soil therefore, we suggest further testing be performed with weathered contaminated soils. Key words: soil washing, biosurfactants, crude oil, contaminated soil, oil removal, oil spill treatment.

The aim of this study was the evaluation of the effects of Aescin + essential phospholipids (AEPL) gel on plasma free radicals (PFRs). In this study, ten patients with venous hypertension, no ulcerations or infections, venous microangiopathy, and varicose veins were treated with topical AEPL applied in a standard protocol. AEPL was applied for 2 weeks, three times daily at the same skin region (perimalleolar internal region at defined distance from the medial malleolus and from the anterior edge of the tibia). In subjects with venous hypertension, PFR values were higher than 420 Carr units. After treatment, the values of PRF decreased to almost normal as an effect of the massage with AEPL gel. The decrease in value was observed in all treatment subjects. A ranking system indicates that the AEPL preparation is active in all subjects who have venous microangiopathy. The decrease in PFR value was greater than 30% in all treatment subjects (p

The aim of this efficacy study was to evaluate the effects of topical aescin + essential phospholipids gel in a histamine model of acute microangiopathy, comparable to microangiopathy observed in chronic venous disease.

The bioavailability of beta-aescin--the main active constituent of horse chestnut seed extract--in a nonretarded test medication in comparison with that in a retarded reference formulation was evaluated in 2 randomized crossover clinical trials involving 18 healthy volunteers each. Serum concentration/time curves derived under steady-state conditions and pharmacokinetic parameters measured during both studies showed no significant difference between absorption rates for the retarded versus nonretarded preparation. In the first study, investigators found a test-to-reference ratio of 1.06 (90% confidence interval [CI] range: 99-113) for the area under the curve (AUC; the primary outcome measure). Absorption rates were diminished during the night compared with daytime rates for both study preparations. In the second study, using AUC and maximum concentration (Cmax) as the primary characteristics, investigators analyzed bioavailability based on the mean of 2 consecutive daytime periods and obtained estimates of 1.07 for AUC (90% CI range: 0.96-1.19) and 1.05 for Cmax (90% CI range: 0.90-1.21). Bioequivalence of the test and reference drug preparations was thus established according to the Note for Guidance on the Investigation of Bioavailability and Bioequivalence. Both treatments were equally well tolerated.