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◍Materials-Hydrolyzed soy protein (hSP):Mixture of peptides
(Average molecular weight 700 Da, Many basic amino acid residues such as Lys,Concentration 20%)TOFUPRO® U, IKEDA CORPORATION
◍ SC collection- The inside of the upper arms by tape-stripping
◍ Carbonylation (induce SCCP)- UV (LAX-C100 solar simulator (Asahi Spectra Co., Ltd.), SQ + UV, Acr
◍ Evaluation of SCCP- Fluorescein-5-thiosemicarbazide (FTSC),
EVOS-FL fluorescence microscope (Thermo Fisher Scientific)
◍Wash out- Soaking the SC in 2 mol/L NaCl
Establishment and application of a novel experimental model of stratum corneum carbonylation induced by UV and sebum
Omoi M1, 2, Arai Y1, Inamura K1, Suzuki N1, Hirao T2
1 IKEDA CORPORATION, MM Park Building 12F, 3-6-3 Minatomirai, Nishi-ku, Yokohama, 220-0012 Japan.2 Faculty of Pharmacy, Chiba Institute of Science, 15-8 Shiomi-cho, Choshi, Chiba, 288-0025 Japan.
Poster-No. P-S2-283
40
50
60
70
80
0 30 120
None SQ
Mean±SDN=3
*: p<0.05Car
bo
nyl
pro
tein
leve
l (A
U)
**
*
UV irradiation (J/cm2)
SCCP was successfully induced by UV irradiation in a dose-dependent manner. We established a novel experimental model of SC carbonylation induced by UV + sebum component.
Promotion of UV-induced SCCP by SQ
We established an experimental stratum corneum carbonyl protein (SCCP) induction model mimicking oxidative modification at the skin surface using
a tape-stripped stratum corneum (SC) irradiated with ultraviolet (UV) + squalene (SQ). This experimental system made it possible to evaluate the efficacy of cosmetic ingredients in suppressing SCCP to shield healthy skin.
Materials and Methods
Conclusion
Results
Loss of the inhibitory effect of hSP on SCCP induction by SQ + UV following removal of hSP
prior to SCCP induction
40
45
50
55
60
65
70
None hSP Hydrolyzedsilk
Lys
Mean± SDN=3
**:p<0.01
Car
bo
nyl
pro
tein
leve
l (A
U)
SQ + UV (120J/cm2)
**
hSP exhibited an excellent inhibitory effect against the SCCP induction as compared with hydrolyzed silk or L-Lys.
Effect of UV irradiation on SQ peroxide value
SQ was irradiated with different amounts of UV. Peroxides were consistently produced in SQ irradiated with UV.
Inhibitory effect of hSP on Acr-induced SCCP induction
SC was treated with deionized water or hSP , then treated without or with 1 mmol/L Acr. hSP showed an excellent inhibitory effect against SCCP induction by Acr.
None No hSP + Acr
Induction of SCCP by UV-irradiated SQ
The SCCP level was increased in a dose-dependent manner by application of UV-irradiated SQ.
SQ with UV
30 J/cm2
SQ with UV
120 J/cm2
SQ without UV
40
45
50
55
60
65
70
75
0 30 120
None hSP
Mean±SDN=4
*: p<0.05
* *
*
Car
bo
nyl
pro
tein
leve
l (A
U)
UV irradiation (J/cm2)
SC was treated without or with hSP, then irradiated with UV + SQ. hSP application to the tape-stripped SC prior to UV + SQ resulted in significant inhibition of SCCP generation.
without UV with UV with SQ + UV
hSP + Acr
Introduction
Typical images of SCCP following treatment without UV, with 120 J/cm2 UV , or 120 J/cm2 UV + SQ.UV promoted greater SCCP induction in the presence of SQ.
DetectionCarbonylationInhibitor Wash outSebum
SC
Observation
SCCPhSP SQ UV 2M NaCl FTSC
Tape-stripping
020406080
0 20 40 60 80 100 120UV irradiation (J/cm2)
Per
oxi
de
valu
e(m
eq/k
g)
Stratumcorneum
hSPUV
Shielded skin
CarbonylationSqualene Aldehydes
UV
Squalene*
*Hydrolyzed soy protein
①Unsaturated lipids (such as SQ)
②Aldehydes
Carbonyl protein (CP) is formed by various oxidative stimuli, including UV radiation.
SC proteins
→ SCCP
We need an experimental system① + ②
→ Lipid peroxides
Various aldehydes (such as acrolein (Acr))Degradation
Targets basic amino acid residues
Include many basic amino acid residues
Pre-wash Post-washhSP + +Wash + -
SQ + UV(30J/cm2)
+ +
Wash - +
No SQ + UV SQ + UV
Detection of carbonyl protein by Acr antibody
SC was treated without or with SQ, then irradiated with UV.Acr adduct was generated in the SC treated with SQ + UV.
without hSP with hSP
Negative control
Inhibitory effect of hSP on UV-induced SCCP induction in the presence of SQ
Inhibitory effect of cosmetic ingredients on UV-induced SCCP induction in the presence of SQ
Removal of adsorbed hSP prior to SCCP induction (pre-wash) eliminated the inhibitory effect of hSP against UV-induced SCCP induction in the presence of SQ.