In vivo oxygen radical generation in the skin of the protoporphyria model mouse with visible light exposure: an L-band ESR study.

Although oxygen radicals are thought to play a key role in the skin injury that is caused by protoporphyria, there is no direct evidence of generation of these radicals in vivo. This study measured the generation of oxygen radicals caused by visible light non-invasively in the skin of griseofulvin-induced protoporphyria model mice, using an in vivo electron spin resonance spectrometer equipped with a surface-coil-type resonator that could detect radicals within about 0.5 mm of the skin surface. A durable nitroxyl radical was administered intravenously as a probe. Light irradiation enhanced the decay of the nitroxyl signal in griseofulvin-treated mice, whereas light irradiation did not enhance the signal decay in control mice. The enhanced signal decay was completely suppressed by intravenous administration of hydroxyl radical scavengers, superoxide dismutase or catalase, or the intraperitoneal administration of desferrioxamine. The enhanced signal decay with illumination was reversible, and quickly responded to turning the light on and off. These observations suggest that the hydroxyl radical is generated via an iron-catalyzed reaction in the skin. This paper demonstrates, for the first time, the specific generation of oxygen radicals in response to light irradiation of the skin of protoporphyria model mice.

Medicamentos según riesgo de desencadenar crisis en pacientes con porfirias agudas / Drugs according to risk of to unchain crisis in patients with acute porphyries

Las variedades agudas de porfirias suelen presentar episodios conocidos como crisis agudas, que pueden ser muy graves y aún fatales. La causa desencadenante más frecuente de estas crisis es el consumo de ciertos medicamentos. Así como algunos fármacos están identificados como productores de estas crisis, otros no tienen ese riesgo. Se puede así distinguir medicamentos sin peligro o "seguros", otros que conllevan el riesgo de desencadenar crisis o "no seguros" y otros en que la información es contradictoria. Esta publicación tiene por objeto facilitar, tanto a pacientes como a médicos tratantes, el listado de principios activos, incluyendo los fármacos que los contienen, ordenados según sean "seguros", "no seguros" y con "información contradictoria" y clasificados según acción farmacológica. A fin de facilitar aún más la selección de medicamentos a usar, se incorpora una segunda lista de productos farmacéuticos que se expenden en el mercado chileno, ordenados también según su riesgo de producir crisis agudas. El mercado de medicamentos es muy dinámico, constantemente están incorporándose nuevos y retirando otros que se dejan de usar, por lo que nuestra Unidad mantiene actualizadas estas listas en nuestra página Web (http://porfiria.med.uchile.cl)

Mdr P-glycoproteins are not essential for biliary excretion of the hydrophobic heme precursor protoporphyrin in a griseofulvin-induced mouse model of erythropoietic protoporphyria.

Hepatic complications in erythropoietic protoporphyria (EPP) have been attributed to toxic actions of accumulated protoporphyrin (PP). PP can only be removed via the bile but transport systems involved have not been defined. The aim of this study was to gain insight in the mode of biliary PP excretion, with emphasis on the potential contribution of the Mdr1 P-glycoprotein export pump and biliary lipids as PP carriers. Control mice and mice homozygous for Mdr1a/b (Abcb1) or Mdr2 (Abcb4) gene disruption, the latter unable to secrete phospholipids and cholesterol into bile, were treated with griseofulvin to chemically induce protoporphyria. All groups showed dramatically increased PP levels in erythrocytes and liver after griseofulvin treatment. Histologically, massive PP deposits were found in livers of control and Mdr1a/b(-/-) mice but not in those of Mdr2(-/-) mice. Serum unesterified cholesterol and phospholipids were increased by griseofulvin because of formation of lipoprotein-X in control and Mdr1a/b(-/-) mice only. Yet, bile flow was not impaired in griseofulvin-treated mice, and biliary bile salt, phospholipid, and cholesterol secretion rates were significantly increased. Surprisingly, biliary PP excretion was similar in all 3 groups of griseofulvin-treated mice: the observed linear relationship between hepatic and biliary PP concentrations and identical liver-to-bile concentration ratios in treated and untreated mice suggest a passive mode of excretion. In conclusion, the data show that Mdr P-glycoproteins are not critically involved in biliary removal of excess PP and indicate that the presence of biliary lipids is required for formation of intrahepatic PP deposits.

Alteration of mRNA levels of delta-aminolevulinic acid synthase, ferrochelatase and heme oxygenase-1 in griseofulvin induced protoporphyria mice.

Human erythropoietic protoporphyria (EPP) is an inherited disorder of porphyrin metabolism and its experimental murine model can be produced by treatment with griseofulvin (GF). We investigated the alteration of mRNA expression in ferrochelatase (FeC), delta-aminolevulinic acid synthase (ALAS) and heme oxygenase-1 (HO-1) in liver, skin and peripheral blood cells of GF-treated mice. In liver, ALAS mRNA was enhanced dramatically by GF administration, in accord with thesis that the expression of ALAS is regulated by feedback mechanism. The expression of HO-1 mRNA increased most rapidly and drastically in liver, however its mechanism of regulation may be different from that of ALAS mRNA. The level of FeC mRNA in liver was less affected with GF treatment. Our results indicate that the inhibition of FeC by GF administration might occur primarily at post-transcriptional level. Similar effects were observed in the ALAS and HO-1 mRNA expression in peripheral blood cells, 2-fold increase in the ALAS mRNA and increase from undetectable level to detectable level in the HO-1 mRNA. In skin of GF-treated mice, average increases of 1.3-fold in the ALAS mRNA and 1.6-fold in the HO-1 mRNA were statistically insignificant. The FeC mRNA level was not altered in peripheral blood or in skin of GF-treated mice. The present study indicates that the molecular analysis is practicable in skin and peripheral blood. In further study, this model could contribute to investigate the pathogenesis of clinical manifestation including possibly cutaneous changes in EPP.

Light-induced deposits in Bruch's membrane of protoporphyric mice.

Photosensitization of choriocapillary endothelium with blood-borne photosensitizers, such as protoporphyrin IX, has been proposed as a mechanism for the choriocapillary sclerosis and Bruch's membrane deposits seen in age-related macular degeneration. Utilizing a mouse model of protoporphyria with approximately a 10-fold increase in protoporphyrin IX level and exposure to blue light (14 microW/cm2; bandwidth, 390 to 430 nm), a time- and light-dependent increase in choriocapillary and sub-retinal pigment epithelium basal laminar-like deposits could be demonstrated at 7 months by transmission electron microscopy. Thickening of the choriocapillary endothelial basement membrane with a homogeneous electron-dense material was first noted in protoporphyric mice exposed to blue light for 13 weeks. At 28 weeks the experimental animals exhibited a thick band of homogeneous deposits at the level of the choriocapillary basement membrane and electron-dense fibrillogranular deposits of varying sizes along the inner aspect of Bruch's membrane, with fibrils measuring up to 16 nm, with a periodicity of 13 nm. These deposits contributed to an overall thickening of Bruch's membrane with narrowing of the choriocapillaris. The morphologic appearance and localization of these deposits within Bruch's membrane of this animal model are similar to previously described deposits noted in the aging Bruch's membrane prior to the development of age-related macular degeneration.