Studies of Photoprotection Against Porphyrin Photosensitization Using Dithiothreitol and Glycerol

Although protection against ionizing radiation by compounds containing sulfhydryl (SN) groups, such as cysteine, has been reported, these agents have been unsuccessful to date in protecting mammals against non-ionizing radiation (>200nm). This study describes successful photoprotection by dithiothreitol (DTT) and glycerol against porphyrin photosensitization having an action spectrum of 400nm. Test models used were red blood cells (RBC) obtained from patients with erythropoietic protoporphyria (EPP) and mice photo-sensitized by hematoporphyrin (HP). A mortality rate approaching a lethal dose in 50% of the animals in 1 day (LD50/24 hrs) was established in 100 white mice that had received an intraperitoneal injection of 100mg HP/kg body wt., and were then irradiated with 5 × 106 ergs/cm2 from a fluorescent light source emitting 320-450nm radiation. Another 100 mice were treated in an identical manner except that they received, in addition to HP, 80mg DTT/kg body weight in a 5.5% glycerol solution. This group showed 75% reduction in mortality (p < 0.03). No lethal effects were observed in animals treated with DTT and glycerol or HP in the above concentrations without 400nm irradiation. RBC obtained from patients with EPP and exposed to 107 ergs/mm2 of 400nm radiation showed 100% hemolysis after 180min. These cells, when irradiated under identical conditions except for the addition of DTT, manifested only 19% hemolysis during this period of time. Measurements of SH groups of RBC from patients with EPP showed a progressive decrease during photohemolysis. Comparison of the rate of photohemolysis of normal and glucose-6-phosphate dehydrogenase (G6PD) deficient RBC irradiated in the presence of protoporphyrin IX revealed that G6PD deficient RBC, which have an impaired ability to produce reduced glutathione (GSH), were more susceptible to porphyrin-induced photohemolysis. These studies demonstrate that DTT and glycerol offer photoprotection in an in vivo mammalian system against porphyrin photosensitization. It is suggested that the mechanism of the photoprotective action against 320-450nm radiation has many features similar to that of radioprotection by thiols and glycerol against ionizing radiation

STUDIES IN PORPHYRIA

Porphyrin biosynthesis in mammalian skin and in skin obtained from patients with selected types of porphyria has been studied. Cutaneous porphyrinogenesis required the precursor δ-aminolevulinic acid (ALA) which, when added to murine, rat, and human skin in vitro, was rapidly converted to porphyrins. Total porphyrin content was quantitated by fluorescence assay, and spectral studies indicated that more than 80% of the porphyrin produced was protoporphyrin. The majority of skin porphyrinogenesis occurred in epidermis or in epidermal derivatives such as hair roots. Known inducers of hepatic δ-aminolevulinic acid synthetase (ALAS), the rate-limiting enzyme for heme biosynthesis, were not inducers when added to skin in vitro.Skin from patients with acute intermittent porphyria demonstrated a 43% decrease in cutaneous porphyrin production as compared to unaffected normals. This is consistent with the known deficiency of uroporphyrinogen synthetase that has been previously demonstrated in the liver and red blood cells of these patients. Porphyrinogenesis in skin of patients with porphyria cutanea tarda was not different from controls.These studies demonstrate that skin has the enzymatic capacity to synthesize porphyrins from added ALA and that cutaneous porphyrinogenesis from ALA is deficient in patients with acute intermittent porphyria

Current Status of Mammalian and Human Models for Predicting Drug Photosensitivity

The status of efforts to develop experimental models for drug photosensitivity reactions in small mammals is reviewed. Tests which are practical and also have a high predictive value in determining photosensitivity hazards to man are the goal of this research. The various animal model systems which have been used are evaluated with respect to these goals

Photoallergic Contact Dermatitis in Guinea Pigs: Improved Induction Technique Using Freund's Complete Adjuvant

Efforts to predict the incidence of photoallergic contact dermatitis in man have been hampered by limitations in the animal which have been developed to date. This study reports an improved induction technique in guinea pigs which correlates well with observed clinical experience in man