Bioactive phytochemicals
A plant and plant-derived technology, applied in the field of Cucurbitaceae extracts, to prepare various compositions used in medicine, can solve problems such as unstable supply, no identity and quality standards, and inaccuracy
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Embodiment 1
[0170] Example 1: inhibition of sialase by idoBR1
[0171] Brief introduction
[0172] Sialidase or neuraminidase is an enzyme that catalyzes the cleavage of terminal sialic acid in oligosaccharides and sugar conjugates. They play an important role in regulating the metabolism of sialic acid-containing molecules in biological systems. They are also virulence factors for many viruses and pathogens such as Tannerella forsythia. Sialidase activity in human neutrophils has been reported to play a key role in host inflammatory responses (Glanz, V.Y, 2019, European J. Pharmacol.842, 345).
[0173] Method. The sialic acidase assay uses 2.8 mM and 0.28 mM inhibitors (or water without inhibitors) and 2.5 nM sialase (including NanH from Forseitanella (T.forsythia) incubated in 0.1 mM methyl umbel-N-acetyl neurine (pH 7.2) in 20 mM sodium phosphate buffer. Terminate the reaction at 30 sec and 60 sec by adding pH 10.5 60 mM sodium carbonate buffer. Quantify the release of fluorescent methyl ...
Embodiment 2
[0176] Example 2: Inhibition of endogenous sialase activity assay values in THP-1 cells
[0177] Brief introduction
[0178]The objective of this study was to determine whether idoBR1 or cucumber extract containing more than 1% idoBR1 (Q-actin batch B17CF001) affects the activity of sialidase in human TTHP-1 (monocyte-like) cell cultures. The results of this study may be a combination of decreased sialase expression or inhibition of idoBR1 on this enzyme.
[0179] method
[0180] Sialase activity tested for THP-1 cell line treatment
[0181] THP-1 cells are cultured in RPMI medium supplemented with mercaptoethanol and glutamine to achieve 80% confluency in the culture flask, and then aspirate the cells and centrifuge at 1500 rpm for 5 min. The cell pellet is then resuspended in 1 ml of RPMI complete medium and routinely counted using a hemocytometer. Incubate cells (5×106) with PMA (10 ng / ml) in separate petri dishes to differentiate THP-1 cells. To determine sialidase activity, ...
Embodiment 3
[0197] Example 3: for measuring cd44-HA (hyaluronic acid) binding activity in the presence of idoBR1 using ELISA Treatment of the THP-1 cell line
[0198] Brief introduction
[0199] CD44 has been shown to be involved in hematopoiesis, homing to mucosal lymphoid tissue, and involved in lymphocyte infiltration into inflamed tissue. The interaction of hyaluronic acid (HA) with CD44 and CD168 (RHAMM) can induce a number of cellular behaviors, including tyrosine kinase, protein kinase C, FAK and PI3K, MAPK, NFκB and RAS, as well as activation of cytoskeletal components required for inflammation and cancer. Although most cells express some form of CD44, not all cells are constitutively bound to HA (Kryworuchko, M. et al., 1999, Cellular Immunol., 194, 54; Nandi et al., 2000, J. Biol.Chem., 275, 14939). Functionally active HA adhesion TO CD44 is produced by inducing sialase by MAPK activation. Studies conducted to understand the role of MAPK in the LPS-induced inflammatory response ha...
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