Determinación de la actividad de la oxido nítrico sintetasa endotelial y del factor NF-kB, en piel de pacientes con Pseudoangiomatosis eruptiva

Abstract

La pseudoangiomatosis eruptiva se caracteriza por la aparición brusca de múltiples pápulas eritematosas, asintomáticas, rodeadas de un halo blanquecino, con remisión espontánea. Histológicamente se observa dilatación vascular con escaso infiltrado inflamatorio. Su etiología permanece incierta a pesar de ser relacionada con virus o picaduras de insectos. Basados en el compromiso vascular, el objetivo del trabajo fue investigar la actividad de la enzima endotelial oxido nítrico sintetasa (eNOS) y la expresión del factor NF-kB por inmunohistoquimica en un intento de esclarecer su patogenia. Material y métodos: Se estudiaron diez pacientes con diagnóstico clínico de pseudoangiomatosis eruptiva (PAE) que presentaron la dermatosis en forma epidémica. Se realizaron biopsias teñidas con Hematoxilina-Eosina y Tricrómico de Masson. Se efectuó estudio virológico de los pacientes Nª 4, 9 y 10 mediante determinaciones serológicas para echovirus, enterovirus, citomegalovirus, parvovirus B19 y hepatitis A, B y C. En cinco pacientes se obtuvo material para determinación de eNOS y NF-kB. Resultados: Todos los pacientes, 5 hombres y 5 mujeres presentaron pápulas eritematosas rodeadas por un halo blanquecino, especialmente en las extremidades, alrededor de las rodillas. Histológicamente mostraron vasos dilatados y células endoteliales prominentes con un infiltrado discreto perivascular. Todos los estudios serológicos fueron negativos. La actividad de eNOS fue significativamente menor comparada con la piel normal (p= 0,002) y la expresión de NF- ĸB fue fuertemente positiva en los vasos de la dermis papilar y reticular. Conclusiones: Todos los pacientes fueron afectados en verano, por lo que la picadura del mosquito debe ser considerada como un factor etiológico. La baja expresión de eNOS está relacionada con la vasodilatación y la expresión aumentada de NF-ĸB confirma que el proceso es de tipo inflamatorio.

Eruptive pseudoangiomatosis (EPA) is a dermatosis characterized by sparse erythematous papules, with vascular appearance and surrounded by a blanching halo. It has acute onset and spontaneous resolution. Histopathology shows vascular affection. Though it has been related to viral infection and insects bites its aetiology and pathogenesis remains unclear. Based on clinical and histological findings, related to vascular affection, the main objective was to observe changes in endothelial nitric oxide synthase (eNOS) activity and nuclear factor-ĸ B (NF-kB) expression related to inflammatory pathogenesis. Materials and methods: Samples from ten patients with clinical diagnosis of EPA were fixed for light microscopy in formalin solution, embedded in paraffin and stained with haematoxylin and eosin and Masson´s trichrome stain. In three patients serological testing for echovirus, enterovirus, cytomegalovirus, parvovirus A and B, hepatitis C and hepatitis B virus were performed. eNOS activity: in five patients snap-frozen tissue was obtained and homogenized in a buffer containing 250 mM sucrose, 50 mM tris-HCl pH 7.4 and added protease inhibitors: SBTI, aprotinin, PMSF, leupeptin, DTT and EDTA. The activity was evaluated in the supernatant measuring the conversion of L-[3H]-arginine in L-[3H]- citruline and subsequent liquid scintillation counting. 4. Expression of NF-ĸB, a promoter of inflammatory processes activated by redox status was determined by immunohistochemistry. Results: The patients, 5 women and 5 men, had 3 to 6 mm, red papules surrounded by a 2 to 5 mm blanching halo, especially located on the lower extremities, around knees, 5 5 forearms and arms. In two patients the lesions were also on the trunk and face. Histological findings show dilated dermal blood vessels with plump endothelial cells and perivascular lymphohistiocytic infiltrate. All serological tests for virus were negative. eNOS activity in lesions was significant lower when compared with normal skin (p= 0,002). NF-ĸB expression was strongly positive in dermal vessels of the papillary and reticular dermis. Conclusions: As all our patients were affected in summer, mosquito bites must be considered as etiologic factor. eNOS lower expression suggests that probably it has no role in this inflammatory process, but linked to blood stasis. The increased expression of NF-ĸB confirms that the inflammatory process is related to the developing process.