La sobreexpresión de poli (ADP-ribosa) polimerasa se correlaciona con el desarrollo de necrosis tubular aguda y con la función precoz del trasplante renal / Poly (ADP-ribose) polimerase overexpression is correlationship with develop of acute tubular necrosis and early function of renal allograft

Poli (ADP-Ribosa) polimerasa (PARP-1) cataliza la ADP ribosilación de proteínas usando NAD(+) como sustrato. La activación de PARP-1 conduce a la depleción intracelular de NAD(+). El daño por isquemia-reperfusión (IR) induce una activación excesiva de PARP-1 y la muerte celular por consumo masivo de ATP. Nuestra hipótesis de trabajo es que la excesiva expresión tubular de PARP-1 en riñones humanos trasplantados es una de las causas directas de la inducción de necrosis tubular aguda (NTA) y contribuye al retraso de la función renal del injerto. Material y Métodos: Estudiamos 193 biopsias de trasplante renal (95 preimplante –biopsia de donante– y 98 postrasplante) incluidas en parafina con diferentes grados de NTA y 65 biopsias renales de donante sin NTA. La NTA se estratificó en cuatro grados: ausente (0); leve (1) [50%]. La expresión nuclear de PARP-1 fue evaluada mediante inmunohistoquímica con el kit de polimeros conjugado con peroxidasa MasVision y el anticuerpo anti- PARP-1 (clón PARP01) y valorada semicuantitativamente de 0 a 3. Resultados: La expresión nuclear de PARP-1 antecedió a los cambios morfológicos sugerentes de NTA. Principalmente la inmunotinción se localizó en los núcleos de células tubulares, cuando fue intensa la lesión también apareció en glomérulos (epitelio de la cápsula de Bowman y células endoteliales de capilares glomerulares). La inmunotición fue observable hasta fases finales de la necrosis tubular. La totalidad de las 95 biopsias renales pre-trasplante con NTA grado 1 (86%) o grado 2 (14%) mostraron expresión nuclear de PARP-1 en túbulos. Las 98 biopsias postrasplante con NTA mostraron expresión más intensa de PARP-1 [grado 2 (45%), grado 3 (25%)]. El grado de NTA se correlacionó significantivamente con la expresión de PARP- 1(r=0,565, p=0,0001, test de Pearson), con una expresión media 2,74±0,45 en los casos de NTA severa frente a 1,94±0,74 en los casos de NTA leve y 0,29±0,45 en los casos sin NTA (p=0,0001, test ANOVA de una vía). En conclusión, PARP-1 está vinculado a la inducción de NTA y desempeña un papel importante en el comportamiento de la función precoz del injerto renal ya que está correlacionada significativamente con el tiempo en recuperar la diuresis eficaz (r=0,578, p=0,0001, test de Pearson) y con los niveles séricos de creatinina en el momento de la biopsia (r=0,649), y a los 3 meses (r=0,363, p=0,0001, test de Pearson) pero no a los 6 y 12 meses postrasplante

Poly (ADP-Ribose) Polymerase (PARP-1) catalyzes ADP-ribosylation of proteins using NAD(+) as substrate. Its overactivation leads to massive NAD+ consumption and ATP depletion. The ischemia/reperfusion injury (IR) induces PARP-1 overactivation and leads to cellular necrosis by massive ATP consumption. Our working hypothesis was that massive PARP-1 tubular expression in allograft human kidneys are a direct cause of acute tubular necrosis (ATN) and contribute to delay in early recovery of renal function (RRF) of the transplanted organ. Material and Methods:A total of 193 paraffin embedded renal allograft biopsies (95 pre-implant –donor biopsies– and 98 post-implant) with several ATN degrees, and 65 control renal biopsies from donors without ATN were studied. ATN degree was classified as: Absence (0); mild (1) [50%]. Nuclear expression of PARP-1 in tubular cells was evaluated by immunohistochemistry using polymer-conjugate MasVision kit and the monoclonal antibody anti-PARP-1 (clone PAR01). It was semiquantitatively determined, and scored from 0 to 3. Results: The nuclear PARP-1 preceded the morphological features of ATN. Immunostaining was located mainly in tubular cells nuclei, in cases of intense injury also was observed in glomeruli (capillary endothelial cells and epithelial cells of Bowman’s capsule). Immunostaining was observed until advanced ATN condition. All 95 pre-transplant renal biopsies with ATN degree 1 (86%) or degree 2 (14%) showed tubular nuclei PARP-1 expression. The 98 post-transplant biopsies with ATN showed more intense expression of PARP-1 [degree 2 (45%), degree 3 (25%)]. Statistically significant relationship between ATN degree and PARP-1 expression was found (r=0.565, p=0.0001, Pearson test), with a mean expression of 2.74±0.45 in sever ATN cases versus 1.94±0.74 in mild ATN cases, and 0.29±0.45 in non-ATN cases (p=0.0001, one way ANOVA test). In conclusion, PARP-1 are linked to induction of ATN, and plays an important role in early graft renal function. This fact is indicated by the stati! stically significant relation with delay in total RRF (r=0.578, p=0.0001, Pearson test), creatinine serum levels at biopsy time (r=0.649) and at 3 months (r=0.363, p=0.0001, Pearson test), but not at 6 or 12 months post-transplant

Increased expression of p38 mitogen- activated protein kinase is related to the acute renal lesions induced by gentamicin

Mitogen-activated protein kinases (MAPK) may be involved in the pathogenesis of acute renal failure. This study investigated the expression of p-p38 MAPK and nuclear factor kappa B (NF-kappaB) in the renal cortex of rats treated with gentamicin. Twenty rats were injected with gentamicin, 40 mg/kg, im, twice a day for 9 days, 20 with gentamicin + pyrrolidine dithiocarbamate (PDTC, an NF-kappaB inhibitor), 14 with 0.15 M NaCl, im, twice a day for 9 days, and 14 with 0.15 M NaCl , im, twice a day for 9 days and PDTC, 50 mg kg-1 day-1, ip, twice a day for 15 days. The animals were killed 5 and 30 days after the last of the injections and the kidneys were removed for histological, immunohistochemical and Western blot analysis and for nitrate determination. The results of the immunohistochemical study were evaluated by counting the p-p38 MAPK-positive cells per area of renal cortex measuring 0.05 mm². Creatinine was measured by the Jaffé method in blood samples collected 5 and 30 days after the end of the treatments. Gentamicin-treated rats presented a transitory increase in plasma creatinine levels. In addition, animals killed 5 days after the end of gentamicin treatment presented acute tubular necrosis and increased nitrate levels in the renal cortex. Increased expression of p-p38 MAPK and NF-kappaB was also observed in the kidneys from these animals. The animals killed 30 days after gentamicin treatment showed residual areas of interstitial fibrosis in the renal cortex, although the expression of p-p38 MAPK in their kidneys did not differ from control. Treatment with PDTC reduced the functional and structural changes induced by gentamicin as well as the expression of p-p38 MAPK and NF-kappaB. The increased expression of p-p38 MAPK and NF-kappaB observed in these rats suggests that these signaling molecules may be involved in the pathogenesis of tubulointerstitial nephritis induced by gentamicin.