Aquaporin Jewellmotz
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E. A. Jewell-Motz, Ph.D., J. R. Kaczvinsky, Ph.D., K. M . Lammers, M.S., S. Xie, Ph.D., R.
P&GBeauty, Cincinnati, OhioUSA
Background
Aquaporins (AQP) are an important class of
proteins that regulate the transport of water
and other small solutes across plasma
membranes. First discovered in the early
1990’s (1), there are now 13 distinct AQP
family members that have been identified in
a variety of mammalian tissues, many of
which are relevant to the consumer products
industry. In particular, two aquaporins
(AQP3 and AQP9) are expressed in human
skin cells (2, 3). AQP3 has been implicated
as playing a key role in the transport and
distribution of epidermal water and glycerol
(3,4) and the regulation of keratinocyte
differentiation (5), both critical processes for
maintaining skin hydration, barrier function
and overall skin health.
● Identify compounds that stimulateaquaporin-3 expression and activity in skin
cells.
● Determine whether identified materials
promote a skin moisturizing benefit in humans
when formulated into product.
Objective
Modulation of Aquaporins to Deliver Consumer Benefitsfor Skin Care
Aquaporin channel
(Science17 October 2003:Vol. 302. no. 5644, pp. 383 –384)
Caffeine Stimulates AQP3Protein Expression in vitro
Caffeine Stimulates AQP3Activity in vitro
ELISA for In Vitro AQP Expression
• Cultured human neonatal keratinocytes were cultured in
96 well plates and treated at 37°C for 48-96 hrs• Fixed cells were incubated with anti-AQP3 antibody.• A peroxidase coupled secondary antibody was added,
and AQP3 protein content was determined after treatment with Horseradish Peroxidase Chromogen
TMB.
Glycerol Transport Assay
● MatTek’s Epiderm cultures were treated with test
compound for 48 hours before adding glycerol to themedium in the lower chamber.
● Glycerol levels within the upper chamber medium weredetermined 24 hours later (Free Glycerol Reagent, Sigma,
St. Louis, MO).
● Measurement in the glycerol assay was converted into apercentage increase over the control (where the control is
equal to 100%). Each mean and SEM were derived from atleast three separate experiments, performed in duplicate.
80
90
100
110
120
130
Control 100uM
%
Increasein
Glycerol
Transport
0.8
0.9
1
1.1
1.2
1.3
1.4
1.5
1.6
Control 100uMFoldChangeinAQPProtein
Product FormuCaffein
Delivers Skin MoBenefit in Hum
Studie
In vivo Expression of AQP 3• 20 Caucasian female subjects, 18-55 years
old• RNA isolated from hip biopsies & 1 leg
biopsy• AQP RNA expression quantitated via
GeneChip™ assay
AQP3 levels decrease withage
20 30 40 50 60 700
2500
5000
7500
10000 r = -0.446
p = 0.063
Age (years)
A Q P 3 S i g n a l
Stratumcorneum
Stratumbasale
Clinical Des●40 female subjects
●4 test sites per leg (8 total per s2 ul/cm2, total 40 ul treatment twweekends- 1x per day
●Assessments performed at basethen 2, 7, and 11 day regression
●Technical measures via expert
dryness and redness,●Panelists were asked not to con
herbal drinks
0
0.1
0.2
0.3
0.4
0.5
0.6
B a s e l i n
e
1 W E E
K
2 W E E
K
3 W E E
K
2
InvivoDrySkinGradeImpro
vements
(vs.moisturize
ralone)
AQP3 is localized tohuman skin