Increased synthesis of high-molecular-weight cPLA2 mediates early UV-induced PGE2 in human skin.
Ultraviolet light (UV) B-induced inflammation is characterized by dramatic increases in prostaglandin E2 (PGE2) synthesis due to enhanced arachidonate deacylation from the membrane. Therefore, the effect of UV on sythesis, mass, and distribution of the high-molecular-weight phospholipase A2 (cPLA2) in cultured human keratinocytes and human skin was studied. The 105-kDa cPLA2 was demonstrated to be the critical enzyme in UV-induced PGE2 synthesis and erythema in the first 6 h postirradiation. Immunoprecipitation of 35S-labeled protein showed cPLA2 synthesis increased three- to fourfold 6 h after irradiation. Immunoprecipitated 32P-labeled cPLA2 demonstrated phosphorylation of cPLA2 was concurrently induced, suggesting that UV also activates cPLA2. This increase in cPLA2 synthesis and activation also closely correlated with increased PGE2 synthesis and [3H]arachidonic acid release and was effectively blocked by both an S-oligonucleotide antisense to cPLA2 and methyl arachidonate fluorophosphate, a specific inhibitor of cPLA2. Biopsy and histochemical examination of erythematous sites expressed increased amounts of cPLA2 whereas nonerythematous irradiated sites did not. In contrast, cyclooxygenase-1 and -2 in cultures and skin explants were unaffected 6 h post-UV, and no change in cyclooxygenase activity was observed at this time. These results suggest that increased cPLA2 synthesis occurs only when skin is exposed to UV doses that are sufficient to cause erythema and indicate expression of cPLA2 participates in acute UV inflammation.
Gresham
A
A
Department of Medicine, Washington University School of Medicine, St. Louis, Missouri, USA.
Masferrer
J
J
Chen
X
X
Leal-Khouri
S
S
Pentland
A P
AP
D000595:Amino Acid Sequence; D000818:Animals; D016718:Arachidonic Acid / Q000378:metabolism; D003600:Cytosol / Q000201:enzymology*; D015232:Dinoprostone / Q000096:biosynthesis*; D004307:Dose-Response Relationship, Radiation; D004890:Erythema / Q000201:enzymology / Q000209:etiology; D006801:Humans; D015603:Keratinocytes / Q000378:metabolism; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010741:Phospholipases A / Q000096:biosynthesis* / Q000737:chemistry; D054467:Phospholipases A2; D011833:Radiation Injuries, Experimental / Q000201:enzymology; D012867:Skin / Q000166:cytology / Q000378:metabolism* / Q000528:radiation effects*; D013997:Time Factors; D014466:Ultraviolet Rays
D016718:Arachidonic Acid; D010741:Phospholipases A; D054467:Phospholipases A2; D015232:Dinoprostone