Hyaluronidase Inhibitory Activity of Polysaccharides Separated from a Fermented Beverage of Plant Extracts

Six anti-allergic phlorotannins from the brown alga Eisenia arborea were examined for their inhibitory effects on oxidation and hyaluronidase activities. To investigate the radical scavenging activities of the phlorotannins, the 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity was evaluated. Although the radical scavenging activity of a typical scavenger, epigallocatechin gallate (EGCg), was the strongest, the six tested phlorotannins exhibited activities of various degrees. When the hyaluronidase inhibitory activity was examined, eckol inhibited the hyaluronidase activity similar to the typical inhibitors, EGCg and disodium cromoglycate. The other five phlorotannins inhibited the enzymatic activity stronger than the above inhibitors. Since the six phlorotannins from E. arborea expressed radical scavenging and hyaluronidase inhibitory activities, E. arborea could be used as an anti-inflammatory and anti-allergic food ingredient.

The genus Ononis L. which belongs to Fabaceae family, is represented by more than 75 species in Europe as well as Atlantic Islands, West Asia and North Africa. The genus is especially distributed closed to the coasts of Mediterranean Sea (Al-Khalil, 1995; Reyes-Betancort & Scholz, 2008). Ononis spinosa L. is a well known species especially in the central region of Turkey and the roots of this species is used as folk medicine against skin diseases due to its wound healing activity (Altanlar et al., 2006; Sever Yilmaz et al., 2006). In this study, in vitro collagenase, elastase and hyaluronidase inhibitory activities of five polyphenolic compounds, namely trifolirhizin, ononin, medicarpin-3-O-glucoside, onogenin-7-O-glucoside and sativanone-7-O-glucoside, were evaluated. The compounds were isolated from the ethylacetate fraction of the roots of O. spinosa and the structure of the bioactive compounds were established by MS, 1H- and 13C-NMR as well as 2D-NMR (COSY, TOCSY, HSQC, HMBC) techniques. According to the results; among the tested compounds, ononin and sativanone-7-O-glucoside were found to possess hyaluronidase and elastase inhibitory activity. At the dose of 100 μg/ml, ononin showed %31.66 hyaluronidase inhibitory activity and %41.75 elastase inhibitory activity; as the results were found to be %45.58 and %46.88 sativanone-7-O-glucoside, respectively.

In Japan, the growing interest in anti-aging skin care is associated with the unprecedented aging society. Skin aging can be attributed to various factors, including the activation of hyaluronidase enzyme in subcutaneous tissues exposed to ultraviolet radiation. This enzyme breaks down hyaluronic acid, leading to skin sagging. Therefore, hyaluronidase inhibitors can effectively prevent skin aging. Previously, food components have been actively explored to search for hyaluronidase inhibitors considering the high safety of these materials. Although lactic acid bacteria (LAB)-fermented foods inhibit this enzyme, their active compounds responsible for hyaluronidase inhibition remain unknown. Thus, in this study, we aimed to explore the mechanism underlying the LAB-mediated inhibition of hyaluronidase activity. Supernatants of a LAB-fermented milk-based beverage were subjected to a hyaluronidase inhibition assay, followed by purification and separation using hydrophobic adsorbents and high-performance liquid chromatography, respectively. Subsequently, liquid chromatograph time-of-flight mass analysis was performed, revealing α-ketoglutarate (AKG) as the inhibitor of this enzyme. The half-maximal inhibitory concentration (IC50) of AKG was approximately 0.13-fold that of the known strong hyaluronidase inhibitor disodium cromoglycate (DSCG). To the best of our knowledge, this is the first report on hyaluronidase inhibition mediated by AKG, a metabolic product of LAB. Additionally, Lactobacillus acidophilus JCM1132 was identified as a highly effective AKG-producing LAB (63.9 µg/mL) through LC-MS/MS-based quantitative analyses using various LAB-fermented milk samples. We anticipate that the findings of this study will potentially support the development of functional foods and cosmetics enriched with AKG.

Radical scavenging and antioxidant activities of Laminaria japonica and fermented its extracts were evaluated. Freeze-dried L. japonica was fermented by Aspergillus oryzae and extracted with distilled water. The extract solution was mixed with ethanol and centrifuged. The supernatant was ethanol soluble fraction, non-polysaccharide fraction (ESF), and residue was ethanol insoluble precipitation, polysaccharide fraction (EIP). ESF was subjected to sequential fractionation with dichloromethane, ethyl acetate, butanol and water. To determine the radical scavenging and antioxidant activities of these, DPPH radical, hydroxyl radical, superoxide anion radical scavenging activities and linoleic acid oxidation were tested. Among the extracts, ESF of fermented L. japonica showed the highest radical scavenging activity. The ESF showed DPPH radical scavenging activity of 64.33% at concentration of 50 ㎍/mL. It was higher than 57.70% of vit. C. Ethyl acetate and butanol fraction had high value of radical scavenging and antioxidant activities, especially butanol fraction of fermented L. japonica was 79.48% of hydroxyl radical scavenging activity at concentration of 50 ㎍/mL. The fermented L. japonica had radical scavenging and antioxidant activities higher than L. japonica. These results suggest that fermented L. japonica is healthy food having radical scavenging and antioxidant activities.

Objective: The present study was undertaken to evaluate the anti-inflammatory, anti-oxidant and antibacterial potential of Boswellia serrata extract. Methods: Antioxidant activity was determined using DPPH, Hydroxyl, and Total antioxidant capacity and nitric oxide radical scavenging activities. Anti-inflammatory activity was determined using hyaluronidase enzyme inhibitory assay in which Indomethacin was taken as a reference drug. Agar diffusion was used as a test for antibacterial activity. Grampositive (Staphylococcus aureus) and gram-negative (Escherichia coli) bacteria were chosen for the experiment. As a standard medication, ciprofloxacin was used. Results: Test sample free radical scavenging activity was assessed at 10μg, 50μg, and 100μg concentrations. Results demonstrated dose-dependent inhibition (75.90% at 100μl concentration). Results for hydroxyl radical scavenging activity were compared to Ascorbic acid at concentrations of 10μl, 50μl, and 100μl. Hydoxyl radical scavenging activity was dosedependent like free radical scavenging activity. Inhibition was highest at 100μl concentration (89%), while standard medication Ascorbic acid demonstrated 94.84%.Sample concentrations (10μg, 50μg, and 100μg) were tested for nitric oxide scavenging activity, with maximal scavenging reported. Obtained 87.64% at 100μl concentration, compared to 89.73% for reference medication BHA. Hyaluronidase enzyme inhibition testing results show 84.84 % inhibition at 100μl concentration (Maximum inhibition). Conclusions: Boswellia serrata standard extract shows dose-dependent free radical scavenging and hyaluronidase inhibitory activities.

This study was designed to examine the nitric oxide production, hyaluronidase, and prostaglandin E2 (PGE2) inhibitory activities for the anti-inflammatory effect and DPPH and ABS radical scavenging activities for antioxidant effect using Pleurotus nebrodensis mycelium extract for a functional tea development. The cell viability was confirmed that all extracts were not cytotoxic to RAW264.7 cell within a concentration of 90 μg/mL irrespective of solvent extracts. The nitric oxide inhibitory activity in LPS-stimulated RAW264.7 cells was the highest at 43.6% using 90 μg/mL of hot water extract. The PGE2 inhibitory level was the highest 32.9% of 90 μg/mL of hot water extract. The hyaluronidase inhibitory activity as the highest 53.5% using 90 μg/mL of hot water extract. On the other hand, in the case of antioxidant activity, the maximum DPPH radical scavenging activity using 90μg/mL of ethanol extract obtained 69.6%. The maximum ABS radical scavenging activity using 90μg/mL of ethanol extract obtained 71.3%. When hot water extract was mixed in green tea, they was considered an excellent extract because the sensory evaluation showed a relatively high value. Based on the results of these experiments, it was possible to confirm the possibility of the functional material development of the tea beverage for anti-inflammatory and antioxidant agents using the hot water and ethanol extract of P. nebrodensis mycelium.

An estimated 20-30% of CML patients will become resistant to tyrosine kinase inhibitors (TKIs) including imatinib. Recent reports suggest that CML resistance to TKI's is driven by interactions with protective microenvironment niches that influence their survival and self-renewal capacity. Hyaluronic acid (HA) has emerged as a key contributor in this phenomenon of CML resistance to TKI’s. HA influences cell viability through molecular weight dependent interactions with cell-surface receptors CD44 and RHAMM. Low molecular weight (LMW) HA, is typically found in association with many cancers including CML and promotes apoptotic resistance, proliferation, and migration of cancer cells. High molecular weight (HMW) HA is primarily produced by fibroblasts and its catabolism to LMW HA is mediated by hyaluronidase (HYAL) activity. HYAL activity may provide a potential therapeutic target in combination with current TKI treatments. Using a co-culture model, we investigated the role of HA produced by bone marrow stromal fibroblasts on CML viability when subject to imatinib (IM) treatment. Furthermore, we investigated the possibility of targeting hyaluronidase as an adjuvant in combination with IM for the treatment of CML. Co-culture with stromal fibroblasts protected CML cells from proliferation inhibition by IM. HA production and its catabolic products including LMW HA were elevated in the supernatant of co-cultures exposed to IM. The expression of genes encoding for HA metabolic proteins including hyaluronic acid synthases and HYALs, were significantly increased in co-cultures subject to IM stress. Additionally, HA treatment of CML mono-culture protected against chemotherapy-induced apoptosis. Treatment of CML co-culture with HYAL inhibitor, glycrrhizic acid (GA), inhibited HYAL activity, greatly reduced proliferation and increased apoptosis of CML cells alone and in combination with IM. This investigation provides additional evidence of the importance of stromal cell support in CML pathology and that resistance to TKI therapy may be mediated in part through upregulation of total HA production and its catabolism. Furthermore, we establish the potent therapeutic effects of GA on CML cells alone and in combination with current IM treatment strategies as a prospective method of circumventing TKI-resistant CML. For the first time we demonstrate GA's effectiveness as a HYAL inhibitor in culture. HYALs are an attractive target not only in CML resistance but in many cancers due to the reported role of its product, LMW HA, in inflammation, proliferation, and apoptotic resistance. Citation Format: Bryan Hostetler, Olga Uchakina, Robert McKallip. Targeting hyaluronidase reduces stromal cell protection of chronic myeloid leukemia to imatinib therapy. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3848.

Hyaluronidase hydrolyzes glycosaminoglycans, including hyaluronan, in the extracellular matrix during tissue remodeling. Hyaluronidase activity increases in chronic inflammatory conditions, e.g., inflammatory joint disease. In this study, we tested the ability of ethanolic extracts (1:9 [wt/vol] of 50% ethanol) of bran from six cultivated varieties of Sorghum bicolor to inhibit hyaluronidase activity in vitro in comparison to extracts of wheat and rice bran. Each extract inhibited hyaluronidase activity with this order of potency: Sumac > Shanqui Red > Black > Mycogen > Fontanelle > White sorghum. Extracts of wheat and rice bran had weak inhibitory activities relative to the high phenolic sorghum brans. Hyaluronidase inhibition correlated positively with total phenolic content and ferric reducing antioxidant power values for each bran extract. Inhibition was not only due to condensed tannins (proanthocyanidins) because the Black sorghum cultivar lacks condensed tannins but has abundant anthocyanins and other polyphenols. Since hyaluronidase activity is important in conditions such as osteoarthritis and skin aging, these sorghum varieties deserve consideration for functional foods and beverages, and for nutraceutical and cosmeceutical ingredients.

The hydrolysates of fresh and boiled Venus clams with five different proteases for the production of low-molecular protein hydrolysates were optimised by response surface methodology. Alcalase hydrolysates exhibited the strongest hyaluronidase inhibitory activity. The optimum hydrolysis conditions of fresh and boiled clams were< enzyme-to-substrate ratio (E/S), 2.15%; time, 150 min; water-to-substrate ratio (W/S), 83.84 mL g− 1 for fresh clam, and E/S, 2.02%; time, 4.11 h; W/S, 69.74 mL g− 1 for boiled clam. The fresh and boiled clam protein hydrolysates were fractionated by S-200 HR size-exclusion chromatography, which resulted in one (FH1) and two (BH1 and BH2) fractions, respectively. BH1 exhibited the highest hyaluronidase and elastase inhibitory activities with specific activities of 141.15 and 81.36% mL mg− 1, respectively. Therefore, the boiled Venus clam hydrolysate might be developed as a cosmeceutical agent because of its strong hyaluronidase and elastase inhibitory activities.

(1) Background: A naturally occurring glycoside, esculeoside B (EsB), has been identified as a major component in juice or canned tomato. We reported how EsB ameliorated mice experimental atopic dermatitis by a decrease in serum IgE levels. However, the underlying immunologic molecular mechanisms are unknown. (2) Methods: The present study tested the effects of EsB on hyaluronidase activity and CD4+ T lymphocyte activation using concanavalin A (ConA)-blast mouse splenocyte primary culture. (3) Results: We found that EsB and its sapogenol esculeogenin B (Esg-B) decreased hyaluronidase activity by a modified Morgan–Elson method. We demonstrated that EsB/Esg-B dose-dependently suppressed T-lymphoproliferation using CFSE-labeled flow-cytometry and water-soluble tetrazolium (WST) assay. Using ELISA and q-PCR methods, EsB/Esg-B suppressed the cytokine secretion and mRNA expression of Th2-relevant IL-4 and Th1-relevant IFN-γ. Moreover, both EsB/Esg-B showed a reduction in IL-10 secretion, but only Esg-B decreased IL-2 secretion. (4) Conclusions: Our study is the first to demonstrate how EsB/Esg-B inhibit hyaluronidase activity and reduce CD4+ T-lymphocyte activation via a reduction in Th2-lymphocyte activity by modulation of Th2/Th1/Treg subunits differentiation.

Hyaluronidase hydrolyzes glycosaminoglycans including hyaluronan (HA) in the extracellular matrix during tissue remodeling. Upregulation of hyaluronidase activity occurs in chronic inflammatory conditions (e.g., inflammatory joint disease). Prior work demonstrated that Vitis rotundifolia. Michx (muscadine) skin extracts possess significant in vitro. and in vivo. anti-inflammatory activities. In this study, we tested the ability of muscadine skin and seed extracts to inhibit in vitro. hyaluronidase activity. Ethanol extracts (1:9 w/v 50% ethanol) were prepared from dried skins and seeds of Early Fry (bronze) and Ison (purple) muscadine varieties. Each extract inhibited hyaluronidase activity. Seed extracts were two to three times more potent per unit weight than skin extracts. The IC50 values of the Ison seed, Early Fry seed, Ison skin, and Early Fry skin were 0.3, 0.6, 1.0, and 1.0 mg/mL, respectively. Hyaluronidase inhibition correlated positively with total phenolic and ferric-reducing antioxidant power (FRAP) values, but not with anthocyanin content. In addition to anti-inflammatory and antioxidant properties, the results of this study show that polyphenolics in muscadine grapes inhibit hyaluronidase. These three bioactivities are important for maintaining healthy connective tissue metabolism.

Cashew nuts are widely eaten, and the juice made from them is a popular healthy drink. In this study 6- [(Z) -8-pentadecenyl] salicylic acid (1) (anacardic acid monoene) was isolated from the shell oil of Indonesian cashew nuts. From this acid, 19 derivatives were synthesized by epoxidation, methylation and/or reduction of functional groups on the aromatic ring. These derivatives were examined for their use as whitening agents which inhibit Tyrosinase activity and hyaluronidase activity, and as scarvengers of superoxide. Compounds (1 b), (4 a), (4 b), (5 a) and (5 b) derived from (1) were found to be inhibited by 7883 % in tyrosinase activity, by 9698 % in hyaluronidase activity. Scavenging activity of superoxid was 6885 %.

BackgroundHyaluronidases have been found as the target enzymes in the development of osteoarthritis (OA) disease. While there is still no curative treatment for this disease, recent studies on the treatment of OA were focused on the effectiveness of natural products which are expected to improve the symptoms with minimal side effects. The aim of this study was to screen selected Malaysian plants on their anti-hyaluronidase activity as well as to evaluate the active plant and its derived fractions on its potential anti-arthritic and antioxidant activities.MethodsA total of 20 methanolic crude extracts (bark and leaf) from ten different plants were screened using a colorimetric hyaluronidase enzymatic assay. The active plant extract (Payena dasyphylla) was then studied for its hyaluronidase inhibitory activity in the interleukin-1β (IL-1β) stimulated human chondrocytes cell line (NHAC-kn) using zymography method. The Payena dasyphylla methanolic bark extract was then fractionated into several fractions in where the ethyl acetate (EA) fraction was evaluated for its inhibitory effects on the HYAL1 and HYAL2 gene expressions using reverse transcription-polymerase chain reaction (RT-PCR) technique. While the MMP-3 and MMP-13 protein expressions were evaluated using western blot method. The phenolic and flavonoid contents of the three fractions as well as the antioxidant property of the EA fraction were also evaluated.ResultsBark extract of Payena dasyphylla (100 μg/ml) showed the highest inhibitory activity against bovine testicular hyaluronidase with 91.63%. The plant extract also inhibited hyaluronidase expression in the cultured human chondrocyte cells in response to IL-1β (100 ng/ml). Similarly, treatment with Payena dasyphylla ethyl acetate (EA) fraction (100 μg/ml) inhibited the HYAL1 and HYAL2 mRNA gene expressions as well as MMP-3 and MMP-13 protein expression in a dose dependent manner. Payena dasyphylla EA fraction has demonstrated the highest amount of phenolic and flavonoid content with 168.62 ± 10.93 mg GAE/g and 95.96 ± 2.96 mg RE/g respectively as compared to water and hexane fractions. In addition, the Payena dasyphylla EA fraction showed strong antioxidant activity with IC50 value of 11.64 ± 1.69 μg/mL.ConclusionThese findings have shown that Payena dasyphylla might contained potential phenolic compounds that inhibiting the key enzyme in osteoarthritis development, which is the hyaluronidase enzyme through interruption of HYAL1 and HYAL1 gene expressions. The degradation of cartilage could also be inhibited by the plant through suppression of MMP-3 and MMP-13 protein expressions. We also reported that the inhibitory effect of Payena dasyphylla on hyaluronidase activity and expression might be due to its anti-oxidant property.

The quality of bread directly depends on several physical and chemical indicators of the quality of flour, which is used in the technology of making products. Various types of flour, phyto-additives, oils, and leavening microorganisms are added to its recipe to improve the quality and increase the biological value of bread. Pure cultures of lactic acid and propionic acid bacteria are among the fermenting microbiota most often used in the technological process of bread production. The work aimed to determine the effect of adding pure cultures of lactic acid and propionic acid bacteria on the technological process of bread production. Three samples of dough and dough were prepared: the first with lactic acid bacteria and yeast; the second – propionic and lactic acid bacteria and yeast; the third only on a yeast base. Then, samples of rye-wheat dough were mixed in these ovens, and bread was baked. The symbiotic effect of propionic acid bacteria on the acidification of the environment, that is, on the development of yeast and lactic acid bacteria, was established because it was sample № 2 in which this entire microbial landscape was present that had the highest acidity of the dough. An interdependent relationship between added bacteria to the wort and reduction in fermentation time was revealed. In test № 2, the time required to reach the final fermentation process was about 77 min, about 30 min faster than in the control test. In dough № 2 with added microbiota, the lifting force time was the lowest. Therefore, the addition of propionic and lactic acid bacteria to the dough at the same time as yeast contributes to the active gas-forming ability of the dough. Thus, introducing this microbiota into the steam will increase semi-finished products in a short production time.

Polysaccharides constitute one of the main groups of wine macromolecules, and the difficulty in separating and purifying them has resulted in them being less studied than other wine macromolecules. In this study, the biological activity of a number of polysaccharide fractions obtained from yeast lees, must, and wine has been analyzed against a large collection of both lactic acid bacteria (LAB) and acetic acid bacteria (AAB) of enological origin. Results showed that a high proportion of AAB strains (60-88%) was inhibited by concentrations lower than 50 mg/L polysaccharide fractions containing intermediate- (6-22 kD) and small-molecular-weight (<6 kD) mannoproteins and oligosaccharide fragments derived from cellulose and hemicelluloses. Results also showed that, in contrast, yeast mannoproteins in concentrations up to 200 mg/L activated the growth of 23-48% of the studied LAB strains when ethanol was present in the culture broth. Specially, yeast commercial mannoproteins of intermediate molecular weight (6-22 kD) were active in increasing Oenococcus oeni growth (81.5% of the studied O. oeni strains) in the presence of ethanol in the culture broth. These effects of wine polysaccharides on bacterial growth provide novel and useful information for microbiological control of wines and winemaking biotechnology.