Eleutherococcus Species Cultivated in Europe: A New Source of Compounds with Antiacetylcholinesterase, Antihyaluronidase, Anti-DPPH, and Cytotoxic Activities.

Secondary metabolites of the roots of Eleutherococcus spp. cultivated in Poland, or the bioactivity, are not fully known. The 75% methanol extracts of five Eleutherococcus spp. (E. senticosus, E. divaricatus, E. sessiliflorus, E. gracilistylus, and E. henryi) were examined for the content of polyphenols and phenolic acids as well as for antiacetylcholinesterase, antihyaluronidase, anti-DPPH∗, and cytotoxic activities. The richest in polyphenols were the roots of E. henryi (10.4 mg/g DW), while in flavonoids the roots of E. divaricatus (6.5 mg/g DW). The richest in phenolic acids occurred the roots of E. henryi [protocatechuic acid (1865 µg/g DE), caffeic acid (244 µg/g DE), and p-coumaric and ferulic acids (55 µg/g DE)]. The highest inhibition of AChE was observed for E. gracilistylus and E. sessiliflorus (32%), at the concentration of 100 µg/0.19 mL of the reaction mixture, while that of Hyal for the roots of E. henryi (40.7%), at the concentration of 100 µg/0.16 mL of the reaction mixture. Among five species tested, the E. henryi extract exhibited the strongest HL-60 cell line growth's inhibition (IC50 270 µg/mL). The extracts reduced DPPH∗ in a time-dependent mode, at the concentration of 0.8 mg/mL. After 90 min from 14.7 to 26.2%, DPPH∗was reduced. A phytochemical composition and activity of the Eleutherococcus species, cultivated in Poland, are still under research; however, on the basis of the results obtained, it may be concluded that they may become a source of phytochemicals and be useful for Europe's citizens.

HPTLC-profiling of eleutherosides, mechanism of antioxidative action of eleutheroside E1, the PAMPA test with LC/MS detection and the structure-activity relationship.

Human body is constantly generating free radicals, which causes oxidative stress. Despite naturally occurring antioxidant systems in human body, free radicals cause lipid, proteins and DNA oxidation. New antioxidants are still urgent as well as their mechanisms of action should be explained. In this study, we investigated the mechanism by which eleutherosides B, E and E1 may act as antioxidants, identified eleutherosides in Eleutherococcus lasiogyne and Eleutherococcus giraldii, and explained in vitro the absorption of eleutheroside E1 based on passive transport. The DPPH∗ and DB-HPTLC tests were used to assess the antioxidant activity. Of the three eleutherosides, only eleutheroside E1 exhibited a strong anti-DPPH∗ activity (EC50 37.03 µg/mL; 63 mMol) compared to the raw extracts (EC50 170 and 180 µg/mL for E. lasiogyne and E. giraldii). This activity was also confirmed by the DB-HPTLC autography technique. According to Zaluski's hypothesis, the antioxidant mechanism of eleutheroside E1 is based on the complexation of DPPH∗ molecule with its aryl radical. During this reaction, the aryl radical of eleutheroside E1 (E1∗) and DPPHH are created. Next, the aryl radical (E1∗) is complexed with another DPPH∗ molecule. Additionally, the aryl radical can be stabilized by the presence of the methoxy groups in the aromatic ring, which increases its antioxidative action. The HPTLC-identification of extracts showed the presence of eleutherosides B, E and E1 in both species. The PAMPA test coupled with LC/MS detection showed a low permeability of eleutheroside E1 across artificial membrane. Because eleutherosides belong to the polyphenols, the TPC and TFC were quantified. The TPC and TFC varied from 51.4 to 49.3 mg/g dry extract for TPC, and from 5.73 to 4.91 mg/g dry extract for TFC, for E. giraldii and E. lasiogyne, respectively. In conclusion, eleutheroside E1 in its pure form could be a chemopreventive ingredient of new pharmacological or dietary products, stimulating the GALT. These findings can explain partially the adaptogenic activity of eleutheroside E1 on the GALT, which has been still unknown.

LC-ESI-MS/MS profiling of phenolics in the leaves of Eleutherococcus senticosus cultivated in the West Europe and anti-hyaluronidase and anti-acetylcholinestarase activities.

Neither secondary metabolites of the spring leaves nor the autumn leaves of Eleutherococcus senticosus species cultivated in Poland, or the bioactivity are known. The richest in polyphenols was the autumn leaves (171.1 mg/g DE), while in flavonoids the spring leaves (107.9 mg/g DE). Using LC-ESI-MS/MS, protocatechuic acid has been identified as the most abundant compound in the spring and autumn leaves (200 and 70 µg/g DE, respectively). Amongst flavonoids, naringenin 7-O-glucoside occurred in the largest amount (20 and 10 mg/g DE in the spring and autumn leaves, respectively). The autumn leaves inhibited Hyal the strongest (74.3%), comparing to the spring leaves (33%). A weak inhibition was found towards AChE (0.64 and 5.8% for the autumn and spring leaves, respectively). To our best knowledge, no information was available on the phytochemical composition and activity of the leaves of E. senticosus cultivated in Poland.

LC-ESI-MS/MS profiling of phenolics from Eleutherococcus spp. inflorescences, structure-activity relationship as antioxidants, inhibitors of hyaluronidase and acetylcholinesterase.

Nature is a source of many plant-based molecules used as pro- or drugs. Eleutherococcus species are native to Asia and the North Russia, and are traditionally used to treat various diseases. In turn, neither secondary metabolites of the species cultivated in the West Europe nor the bioactivity is known. No differences in the phenols and flavonoids content in the inflorescences were found. The richest in polyphenols was E. giraldii (5.18 mg/g), while in flavonoids it was E. gracilistylus (1.80 mg/g). Using LC-ESI-MS/MS, protocatechuic and trans-caffeic acids have been identified as the most abundant compounds in E. gracilistylus, E. giraldii, E. senticosus (833.4; 855.6; 614.7 and 280.8; 156.0; 167.6 µg/g DE). It was observed that all species were able to chelate Fe2+ with the EC50 value of 0.2, 0.6, 0.3 mg/mL for E. gracilistylus, E. giraldii, E. senticosus, respectively. E. gracilistylus exhibited the strongest antiperoxidation and anti-DPPH∗ activity (EC50 3.2 and 0.48 mg/mL). The weak inhibitory potential has been observed in case of AChE inhibition at the level of 16.17 and 12.2% for E. gracilistylus, E. giraldii. We report for the first time that the extracts inhibited Hyal activity in the range from 16.4 to 60.7%. To our best knowledge, no information was available on this activity of the inflorescences and this provides a background to study inflorescences in more detail. Considering the SAR, an antioxidant activity may be correlated with a high amount of protocatechuic and trans-caffeic acids and their chemical structure.

Phytoconstituents and Nutritional Properties of the Fruits of Eleutherococcus divaricatus and Eleutherococcus sessiliflorus: A Study of Non-European Species Cultivated in Poland.

Eleutherococcus fruits have been consumed in Russia and Asia throughout the centuries. Currently, there is an increasing interest in these products by the community of Western Europe. Many people suffer from micronutrient deficiencies, known as malnutrition, which consequently influences body condition. The aim of this study was to investigate pharmaconutrition, proximate, mineral, and fatty acid composition, total phenolics content, and total flavonoids content of Eleutherococcus divaricatus and Eleutherococcus sessiliflorus fruits cultivated in Poland. Eleutherococcus divaricatus and E. sessiliflorus contain a high amount of protein and fibres (16.70% and 12.28%; 61.41% and 45.63%, resp.). The fruits were generally high in K (21 g/kg) and low in sodium (0.001 g/kg). In terms of fatty acid composition, both species had a high amount of monounsaturated fatty acids (54.84-57.95%) and polyunsaturated fatty acids (36.22-37.0%). Using LC-ESI-MS/MS, protocatechuic acid has been identified as the most abundant compound, ranging from 260 to 810 µg/100 g DE. Among flavonoids, hyperoside was found to be in the highest amount (120-780 µg/100 g DE). Considering a rich chemical composition of the fruits, a better understanding of their health benefits is important in order to increase their utility and to enrich dietary sources of health promoting compounds. Because of a high amount of protein and a low calorific value, the fruits should be considered food for vegans or vegetarians.

Phytochemical Content and Pharma-Nutrition Study on Eleutherococcus senticosus Fruits Intractum.

In the past two decades public interest in herbal products has increased significantly in Europe, especially in the plant-based products from non-European traditions. Eleutherococcus senticosus has been used for the treatment of inflammatory diseases, anemia, and rheumatoid arthritis. The Eleutherococcus senticosus fruits intractum was examined for the content of phenolic acids (LC-ESI-MS/MS), minerals (AAS), TPC, and TFC (spectrophotometric assay). The antioxidant activity was determined using free radical scavenging assay and TLC-DB-DPPH∗ dot-blot test. An anti-Hyal activity was evaluated by the spectrophotometric assay method. Cytotoxicity towards HL-60, HL-60/MX1, HL-60/MX2, CEM/C1, and CCRF/CEM leukemic cell lines was done using trypan blue test. Among eight phenolic acids, trans-caffeic acid was found in the largest amount (41.2 mg/g DE). The intractum presented a high amount of macroelements (Ca, Mg, K; 1750, 1300, and 21000 mg/kg) and microelements (Fe, Mn; 32.7, 54.3 mg/kg), respectively. The content of TPC and TFC was 130 and 92 mg/g DE, respectively. The intractum showed anti-Hyal activity (2.16-60%) and an antioxidant capacity (EC50; 52 µg/mL). The intractum most strongly inhibited the growth of HL-60, HL-60/MX1, and CCRF/CEM. A better understanding of the intractum health benefits is important in order to increase its utility and enrich dietary sources of health promoting compounds.

In Vitro Anti-AChE, Anti-BuChE, and Antioxidant Activity of 12 Extracts of Eleutherococcus Species.

Neurodegenerative diseases are one of the most occurring diseases in developed and developing countries. The aim of this work focused on the screening of the natural inhibitors of AChE and BuChE and antioxidants in Eleutherococcus species. We found that the ethanol extracts of E. setchuenensis and E. sessiliflorus showed the strongest inhibition towards AChE (IC50: 0.3 and 0.3 mg/mL, resp.). Among chloroform extracts, the most active appeared to be E. gracilistylus (IC50: 0.37 mg/mL). In turn, the ethanol extract of E. henryi inhibited the strongest BuChE with IC50 value of 0.13 mg/mL. Among chloroform extracts, E. gracilistylus, E. setchuenensis, and E. sessiliflorus appeared to be the strongest with IC50 values of 0.12, 0.18, and 0.19 mg/mL. HPTLC screening confirmed the presence of inhibitors in extracts. All extracts exhibited anti-DPPH⁎ activity and single antioxidants have been identified. To the best of our knowledge, no information was available on this activity of compounds in Eleutherococcus. These studies provide a biochemical basis for the regulation of AChE and BuChE and encourage us to continue isolation of active compounds.

Activity of cathepsin D and α1-antitrypsin in blood of men with malignant melanoma.

INTRODUCTION: The aetiopathology of melanoma is (like in other cancers) a result of environmental factors and genetic predispositions. One of the most important environmental factors which has an influence on melanoma aetiology is exposure to UV radiation. AIM: To estimate the activity of selected parameters in melanoma diagnosis. MATERIAL AND METHODS: In serum of 25 men with melanoma, activity of cathepsin D and α1-antitrypsin (important inhibitors of protease) was estimated. It was a heterogeneous group, because some men were classified according to Clark's classification, and other men were classified by Breslow's classification. Three months after resection of the lesion, only 13 men came back for the next tests. RESULTS: Activity of α1-antitrypsin in patients before and after removing the affected tissue was similar and related to the control value (0.78 mg of trypsin). Medium activity of cathepsin D in the control group was about 11.88 nM. Activity in patients before surgery was lower: 8.94 nM. After removing the tumour, this activity was 9.31 nM. CONCLUSIONS: It can be supposed that the proteolytic-antiproteolytic balance was disturbed in persons with melanoma, because the activity of cathepsin D in serum of these people was lower than in healthy people, but the activity of α1-antitrypsin was not changed.

[Arylsulfatase activity in the blood of men with malignat melanoma. Preliminary reports]. / Aktywnosc arylosulfatazy we krwi mezczyzn z czerniakiem skóry.

UNLABELLED: High arylsulfatase activity occurs in the course of cutaneous melanoma. There is a correlation between the activity of the enzyme, and the susceptibility to metastatic cancer, including metastatic melanoma. Melanoma of the skin were 1.4% of all cancers in men and the number of deaths from this cancer in Poland in 2002 was the 424th. The aim of the study was decided to determine the usefulness of arylsulfatase activity in patients with melanoma of men for diagnostic purposes and for monitoring of the disease course. MATERIAL AND METHODS: The study included 25 patients diagnosed men with melanoma (mean age 55.5 years). Blood samples were collected before excision of melanoma of the skin and in the period from 2 to 3 months after surgery. To donate blood after resection of malignant melanoma reported only 13 men (mean age 57.4 years). Arylsulfatase activity was determined by Roy the modified Bleszczynski method. RESULTS: Arylsulfatase activity before surgery to remove the changes was higher in comparison to the control group and the patients after surgery. CONCLUSIONS: Of men patients with malignat melanoma arylsulfatase activity was high. Arylsulfatase mark may be useful in diagnosing and monitoring the course of the disease.