8/8/2019 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