74 MARINE INGREDIENTS TNF-α■ IL-6 ■
1.2 1.0 0.8 0.6 0.4 0.2 0
Ctrl -46.4% -51.4%
1.2 1.0 0.8 0.6 0.4 0.2 0
Ctrl
Red grape algae extraction 0.2% Quercetin
0.005%
Figure 1: Effect of red grape algae extract on the β-hexosaminidase secretion of RBL-2H3 cells (Ctrl = control)
In vitro studies Antiallergic activity: materials and methods RBL-2H3 cells, rat basophilic leukemia mast cells, are commonly employed in inflammation, allergy and immunological research and has proven to be a pertinent model for skin allergy. Upon activation of RBL-2H3 cells, degranulation occurs, leading to the release of markers such as β-hexosaminidase. This model mimics the pathophysiology
of skin allergy in vivo. A23187, a calcium ionophore, was used to activate the release of β-hexosaminidase from RBL-2H3 cells. Quercetin is a control group to suppress β-hexosaminidase activity as an index of anti- allergy activity. RBL-2H3 cells were seeded in 96-well plates. After an overnight incubation, those
RBL-2H3 cells were treated with 0.05% and 0.2% red grape algae extract and quercetin. Then the cells were stimulated with A23187. Released β-hexosaminidase was detected by measuring optical absorbance at 405 nm.
Results Figure 1 shows that RBL-2H3 cells were stimulated with A23187, which directly evokes degranulation, leading to the release of
-97.0%
-93.1%
0.2% Red grape algae extraction
0.1% 0.05%
Figure 2: Secretion of proinflammatory TNF-α and IL-6 in LPS-stimulated cells after treatment with 0.2%, 0.1% and 0.05% red grape algae extract. TNF-α: tumor necrosis factor-alpha; IL-6: interleukin-6; LPS: lipopolysaccharide (Ctrl = control)
intracellular β-hexosaminidase from RBL- 2H3 cells. Pretreatment with 0.05% and 0.2% red grape algae extract significantly reduced release of β-hexosaminidase from cells. The inhibition ratios are 46.4% and 51.4% respectively.
Anti-inflammatory activities: materials and methods Macrophages are specialized phagocytic cells of the innate immune system and play an important role in inflammatory responses. Therefore, in many studies, macrophages have been utilized as a cell type for immune response experiments. Lipopolysaccharide (LPS) is the
component of Gram-negative bacteria cell walls and can induce an inflammatory reaction by triggering the release of inflammatory cytokines, including interleukin-6 (IL-6) and tumor necrosis factor α (TNF-α). Macrophages were seeded in 96-well
plates. After an overnight incubation, those macrophages were treated with 0.05%, 0.1% and 0.2% red grape algae extract. Then the cells were stimulated with LPS. The levels of IL-6 and TNF-α in the cell culture supernatants were detected using ELISA kit.
Results Figure 2 shows the effects of red grape algae extract exposure on the LPS-stimulated IL-6 and TNF-α production of macrophages. IL-6 and TNF-α production was significantly downregulated in a dose-dependent manner in macrophages treated with red grape algae extract compared to the control group. Specifically, 0.2% red grape algae extract inhibited 93.1% and 97.0% of IL-6 and TNF-α production respectively in the macrophages.
Anti-ageing effect: materials and methods The proliferation of fibroblasts and keratinocytes plays the major role in the skin repair process and skin ageing. Therefore, fibroblasts and keratinocytes are commonly used to skin related studies. Fibroblasts and keratinocytes were seeded in 96-well plates. After an overnight incubation, those cells were treated with 0.025%, 0.05%, 0.01% and 0.2% red grape algae extract for 24 hours. The MTS/PMS assay is used to assess cell proliferation.
Results Figure 3 shows red grape algae extract significantly stimulate fibroblasts and
150 125 100 75 50 25 0
Ctrl
Increased 23%
150 125 100 75 50 25 0
0.025% 0.05% 0.1% 0.2% Ctrl
Increased 23%
0.05%
0.1%
0.2% Figure 3: The proliferation of fibroblasts and keratinocytes after treatment with 0.025%, 0.05%, 0.1% and 0.2% red grape algae extract (Ctrl = control) PERSONAL CARE March 2024
www.personalcaremagazine.com
Fibroblast viability %
β-Hexosaminidase OD 405
Keratinocyte viability %
Inflammatory cytokines (%)
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