Identification of pancreatic type I secreted phospholipase A2 in human epidermis and its determination by tape stripping.
Phospholipases A2 (PLA2) catalyse the release of fatty acids from the sn-2 position of phospholipids and have been suggested to play a key part in permeability barrier homeostasis. Using a sensitive and versatile fluorometric method, significant PLA2 activity has been detected in both human skin homogenates and tape strippings of stratum corneum. Based on various properties (resistance to heat and sulphuric acid treatment, neutral optimal pH, absolute requirement for millimolar calcium concentrations, inhibition by dithiothreitol and p-bromophenacyl bromide, and resistance to a trifluoromethyl ketone derivative of arachidonic acid, AACOCF3, a specific inhibitor of cytosolic PLA2), this enzyme was characterized as a secretory PLA2 (sPLA2). Immunohistochemistry revealed strong labelling of type I pancreatic sPLA2 at the stratum corneum-stratum granulosum junction, type II sPLA2 being undetectable. An increase in PLA2 activity in tape-stripped material from the deepest level of the stratum corneum was correlated with partial morphological disappearance of type I sPLA2 immunolabelling. Our data thus provide the first convincing evidence that pancreatic sPLA2 is significantly expressed in human epidermis, where it might participate in the accumulation of free fatty acids contributing to the permeability barrier. In addition, our method for determining PLA2 activity in easily available tape strippings should allow further clinical studies aimed to explore possible PLA2 abnormalities in various dermatoses.
Mazereeuw-Hautier
J
J
Service de Dermatologie, Hôpital de Rangueil, 1 avenue J Poulhès, 31403 Toulouse Cedex 4, France.
Redoules
D
D
Tarroux
R
R
Charveron
M
M
Salles
J P
JP
Simon
M F
MF
Cerutti
I
I
Assalit
M F
MF
Gall
Y
Y
Bonafe
J L
JL
Chap
H
H
D000293:Adolescent; D000328:Adult; D001706:Biopsy / Q000379:methods; D002118:Calcium / Q000378:metabolism; D004817:Epidermis / Q000201:enzymology*; D005260:Female; D006801:Humans; D007150:Immunohistochemistry; D008875:Middle Aged; D010741:Phospholipases A / Q000737:chemistry* / Q000302:isolation & purification; D054467:Phospholipases A2
D010741:Phospholipases A; D054467:Phospholipases A2; D002118:Calcium