ANTI-POLLUTION 25 The expression of NQO1 was also


greatly enhanced (+47%) as observed using a western blot assay (Fig 4c).


Control BaP


Red Vine Leaf Extract protects skin cells from BaP In order to evidence that Red Vine Leaf Extract can protect skin cells because of its ROS scavenging properties, and because of the enhancement of the cell detoxification system, keratinocytes were pretreated with Red Vine Leaf Extract and then exposed to BaP. Monitoring of DNA damages was performed by the detection of the phosphorylated histone gH2Ax, a useful marker to detect various types of DNA damages.10


While an exposure to the BaP


led to a dramatic increase in DNA alterations, the pretreatment with Red vine Leaf Extract led to a 68% decrease in the amount of damaged DNA (Fig 5).


Sulforaphane


140 120 100 80 60 40 20 0


Control BaP (2.5µM) Redivine *** *** -68%


Red Vine Leaf Extract and indoor pollution l Red Vine Leaf Extract protects skin against house dust Cultured keratinocytes were exposed to a standardised house dust (NIST-SRM 2584) for 24h in the presence or in the absence of Red Vine Leaf Extract (Fig 6). While exposure to house dust led to a 29% decrease in cell viability, treatment with Red Vine Leaf Extract (0.05%) reduced the cell mortality down to 14.1%, hence preventing 52% of house dust-induced cell death. Red Vine Leaf Extract is therefore capable of efficiently opposing the noxious effects of house dust.


Sulforaphane (5µM) Redivine (0.1%)


Figure 5: Redivine protects skin cell DNA from pollution. Immunostaining pictures of keratinocytes exposed to BaP (2.5µM) that were treated with Redivine or sulforaphane. Damaged DNA (in green) was quantified. Cell nuclei are in blue (DAPI). (vs. BaP: ***p-value <0.005)


Red Vine Leaf Extract activates the Nrf2 pathway In order to assess the Nrf2 pathway activation, cultured keratinocytes were treated with Red Vine Leaf Extract. Nrf2 translocation from the cytoplasm to the nucleus (in the absence of any external stress) was monitored by immunofluorescence (Fig 4a). Sulforaphane, a well-known pharmaceutical activator of Nrf2, was used as a positive reference. The immunofluorescence assay shows well the Nrf2 translocation to the nucleus


Table 1: Redivine has strong anti-oxidant properties. The anti-oxidant properties were assessed using DPPH, ABTS and PNBT techniques.


EC50


DPPH ABTS PNBT


November 2018 (%)


0.071 ± 0.012 0.052 ± 0.018 0.075 ± 0.09


with a 153% higher nuclear location (Fig 4a) after the treatment with Grape Leaf Extract. Translocation was comparable when cells were exposed to the positive reference sulforaphane. To verify that Nrf2 translocation was followed by a downstream over-production of protective mediators (cf. Fig 2), we checked for glutathione (GSH) and NAD(P)H:quinone oxidoreductase 1 (NQO1) expression after treatment with Red Vine Leaf Extract. These two mediators are endogenous antioxidants and thus play an important role in BaP detoxification. Besides, both mediators are involved in BaP metabolites deactivation.8,9 Measurement of intracellular GSH content was assessed by fluorimetry. It confirmed that Red Vine Leaf Extract enhances the production of this powerful antioxidant molecule (Fig 4b). This enhancement was comparable with the positive reference.


l Red Vine Leaf Extract protects skin against blue artificial light


Artificial light, and especially blue light, may affect skin and cause premature ageing. Indeed, it was reported to cause skin hyperpigmentation, to delay skin barrier recovery and to produce free radicals, especially ROS.3 In order to assess Red Vine Leaf Extract’s ability to protect skin against the noxious effects of domestic light, fibroblasts were exposed to domestic blue light (380-470nm – 30J/cm2) for 24h in the presence or in the absence of Red Vine Leaf Extract. N-acetyl cysteine (NAC), a potent reference anti- oxidant, was used as the positive reference. As ROS production induced by visible light was reported to stimulate the production of matrix metalloproteinase-1 (MMP-1), an enzyme responsible for collagen degradation,11


the production of both


MMP-1 (ELISA test) and type 1 collagen (cytoimmunofluorescence) by cultured human fibroblasts exposed to domestic blue light was monitored. As expected, blue light exposure induces a dramatic increase in MMP-1 expression and a concomitant reduction of detectable collagen (Fig 7a, b). The


PERSONAL CARE EUROPE


Damaged DNA (% of BaP)


Page 1  |  Page 2  |  Page 3  |  Page 4  |  Page 5  |  Page 6  |  Page 7  |  Page 8  |  Page 9  |  Page 10  |  Page 11  |  Page 12  |  Page 13  |  Page 14  |  Page 15  |  Page 16  |  Page 17  |  Page 18  |  Page 19  |  Page 20  |  Page 21  |  Page 22  |  Page 23  |  Page 24  |  Page 25  |  Page 26  |  Page 27  |  Page 28  |  Page 29  |  Page 30  |  Page 31  |  Page 32  |  Page 33  |  Page 34  |  Page 35  |  Page 36  |  Page 37  |  Page 38  |  Page 39  |  Page 40  |  Page 41  |  Page 42  |  Page 43  |  Page 44  |  Page 45  |  Page 46  |  Page 47  |  Page 48  |  Page 49  |  Page 50  |  Page 51  |  Page 52  |  Page 53  |  Page 54  |  Page 55  |  Page 56  |  Page 57  |  Page 58  |  Page 59  |  Page 60  |  Page 61  |  Page 62  |  Page 63  |  Page 64  |  Page 65  |  Page 66  |  Page 67  |  Page 68  |  Page 69  |  Page 70  |  Page 71  |  Page 72  |  Page 73  |  Page 74  |  Page 75  |  Page 76  |  Page 77  |  Page 78  |  Page 79  |  Page 80  |  Page 81  |  Page 82  |  Page 83  |  Page 84  |  Page 85  |  Page 86  |  Page 87  |  Page 88  |  Page 89  |  Page 90  |  Page 91  |  Page 92  |  Page 93  |  Page 94  |  Page 95  |  Page 96  |  Page 97  |  Page 98  |  Page 99  |  Page 100  |  Page 101  |  Page 102  |  Page 103  |  Page 104