The Effect of Temperature on Photosensitization of Erythrocyte Hemolysis by Protoporphyrin and Evaluation of Erythrocyte/ Protoporphyrin/ Human Serum Albumin System

Abstract

Objectives: In this study, we aimed to investigate the effect of temperature on photosensitization of erythrocyte hemolysis by protoporphyrin. We also investigated absorption and fluorescence spectra of erythrocyte/protoporphyrin/human serum albumin system. Material and Methods: Delayed photohemolysis measurements were made in photosensitized erythrocytes by protoporphyrin at irradiation temperatures from 5°C - 35°C and post-irradiation incubation temperatures from 5°C - 42°C. By measuring absorption and emission spectra of protoporphyrin on erythrocyte and human serum albumin (HSA) solutions, erythrocyte/protoporphyrin/HSA system was investigated. Results: Delayed photohemolysis were accelerated at higher irradiation temperature and higher post-irradiation incubation temperature in photosensitized erythrocytes by protoporphyrin. By using spectrometric investigations, strong emission band at 632 nm and a weaker band near 700 nm were determined on erythrocyte/protoporphyrin/HSA system. A 648 photoproduct band was obtained when protoporphyrin was irradiated in the presence of HSA without erythrocyte and this band continued to decrease with increasing irradiation. The formation of the 648 nm band with HSA alone strongly suggests that protoporphyrin bound to erythrocyte reacts with HSA in the buffer. Conclusion: Delayed photohemolysis rate were increased with the square of the light dose over this range of temperature. These temperature dependent photohemolysis curves are consistent with a kinetic model based on multihit target theory. The spectral and photohemolysis properties of the erythrocytes/protoporphyrin/HSA system have been of interest in connection with erythrocyte from erythropoietic porphyria patients. Key words: Protoporphyrin, Photosensitization, Human serum Albumin, Erythrocyte

Keywords

• Protoporphyrin, Photosensitization, Human serum Albumin, Erythrocyte

Protoporfirinle Fotosensitize Edilmiş EritrositlerdekiHemoliz Mekanizmasına Sıcaklığın Etkisi Ve ritrosit/ Protoporfirin/ İnsan Serum Albümini Sisteminin İncelenmesi+

Öz

Amaç: Bu çalışmalarda protoporfirinle fotosensitize edilmiş eritrositlerdeki hemoliz mekanizmasına sıcaklığın etkisi ile eritrosit/protoporfirin/insan serum albümini sisteminin soğrulma ve floresans spektrumlarının incelenmesi amaçladı. Gereç ve Yöntem: Protoporfirinle fotosensitif hale getirilen insan eritrositleri, çeşitli sıcaklıklarda (5-35°C) görünür ışına maruz bırakılıp, takiben karanlıkta değişik inkübasyon sıcaklıklarında (5-42°C) bekletilerek oluşan gecikmiş fotohemoliz ölçümleriyle birlikte protoporfirinin eritrosit ve serum albümin solüsyonundaki soğrulma ve emisyon spektrumları ölçülerek eritrosit/protoporfirin/insan serum albümini sistemi incelendi. Bulgular: Protoporfirin ile fotosensitize edilmiş eritrositlerde görünür ışık uygulanması ve takip eden karanlık devredeki sıcaklıkların artmasıyla, gecikmiş fotohemolizin hızlandığı görüldü. Spektrometrik incelemelerde eritrosit/ protoporfirin/ insan serum albümini sisteminde 632 nm'de kuvvetli ve 700 nm civarında ise zayıf emisyon bandı izlendi. Protoporfirin ile insan serum albümini bulunan ortamda eritrositlerin varlığından bağımsız olarak 648 nm'de oluşan foto-ürün, uygulanan ışık dozu arttıkça azalmaktadır. Bu 648 nm'deki emisyon bandının varlığının eritrositlere bağlı protoporfirinin tampon içinde insan serum albüminiyle reaksiyona girdiğini gösterdi. Sonuç: Gecikmiş fotohemoliz hızı ışık dozunun karesine olan ilişkisinin sıcaklık değişimi sonucu arttığı ve sıcaklığa bağımlı fotohemoliz eğrilerinin “Çok Vuruşlu Hedef Teori'' ile uyumlu olduğu saptandı. Eritrosit/protoporfirin/insan serum albümini sisteminin spektral ve fotohemolitik özellikleri, eritropoietik porfiri hastalarının eritrositleriyle benzerlik gösterdiği belirlendi. Anahtar kelimeler: Protoporfirin, Fotosensitizasyon, İnsan serum albümini, Eritrosit

Anahtar Kelimeler

• Protoporfirin, Fotosensitizasyon, İnsan serum albümini, Eritrosit

Kaynakça

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