Association and methods for fighting against the signs of ageing
A technology of signs, compositions, applied in the field of cosmetic treatments, capable of addressing the problems of loss, wrinkles, fine lines skin elasticity etc.
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Embodiment 1
[0129] Example 1: The effect of the combination of betaine and trans-resveratrol on ATP production
[0130] Place human fibroblasts at 37°C, containing 5% CO 2 Cultured on MEM supplemented with glucose, serum and glutamine in a humid atmosphere. Add the solution containing trans-resveratrol and / or betaine to the medium:
[0131] The primary fibroblast cell culture was incubated with resveratrol at a final concentration of 1 μM, betaine at a concentration of 1 mM, or a mixture of both for 120 minutes.
[0132] Cell viability was measured after incubation by fluorescence. Measure intracellular ATP concentration by chemiluminescence-based assay; take the average of 4 measurements.
[0133] The intracellular ATP concentration is expressed in μM of ATP per living cell, and the percentage of ATP increase is calculated relative to the ATP level in untreated cells.
[0134] The results are shown in the attached drawings and provided in the table below.
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[0137] The combinatio...
Embodiment 2
[0140] Example 2: Resveratrol and betaine increase mitochondrial mass
[0141] Place human fibroblasts at 37°C, containing 5% CO 2 Cultured on MEM supplemented with glucose, serum and glutamine in a humid atmosphere. In the presence or absence of the following compounds, the cells were treated with chemical reagents (FK866): trans-resveratrol and betaine. The chemical agent FK866 reduces the amount of cell mitochondria, thereby mimicking one of the signs of aging.
[0142] Under these conditions, the ability of the active ingredients to increase mitochondrial mass was evaluated.
[0143] After 24 hours of treatment, a nonylacridine orange (NAO, Invitrogen) probe that fluoresces at 533 nm was used to measure the amount of mitochondria. Cell viability can be measured by DAPI staining. FACS LSRII was used for flow cytometry analysis, and the results were analyzed by FlowJo software.
[0144] The results are shown in the following table:
[0145] a) Treatment with FK866 reduces the amou...
Embodiment 3
[0150] Example 3: Resveratrol and betaine increase collagen production
[0151] Skin fibroblasts were cultured with resveratrol, betaine or a mixture of the two compounds for 24 hours.
[0152] For gene expression analysis, total RNA was extracted from fibroblast cell culture, and expression was measured by quantitative real-time PCR. RNA extraction was performed using RNeasy Mini Kit (Qiagen). The total RNA was used for cDNA synthesis using iScript cDNA synthesis kit (Biorad). Use mix 480SYBR Green I Master (Roche) performs PCR reaction. Perform two PCRs on each sample using the appropriate primers listed in the table below. In order to compare relative expressions, the ΔΔCt method is used. The relative expression of the gene of interest is represented by the expression variation compared to the untreated sample.
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[0155] The results are shown in the following table:
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[0157] Resveratrol and betaine have a synergistic effect on increasing the expressi...
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