ABSTRACT
OBJETIVO: Determinar a concentração de nitrato/nitrito no líquido cefalorraquidiano e no interstício do corno dorsal entre L6-S1 da medula espinhal em ratas com ou sem cistite induzida por ciclofosfamida. MÉTODOS: Todos os experimentos foram conduzidos usando ratas Wistar. Um probe de microdiálise foi implantado no espaço subaracnoide ou no tecido da medula espinhal nos segmentos L6-S1 (confirmado histologicamente). Dois dias depois, o probe de microdiálise foi perfundido com líquido cefalorraquidiano artificial, contendo ou não NG-monometil-L-arginina. As amostras foram coletadas a cada 15 minutos e mantidas a -20ºC. As concentrações de nitrito/ nitrato foram determinadas por quimiluminescência. RESULTADOS: Nos animais normais, os valores médios das concentrações de nitrito/nitrato, na primeira amostra de microdialisado de líquido cefalorraquidiano e do interstício da medula espinhal, foram semelhantes (482,5±90,2pmol/75µL, n=20, e 505,7±11,5pmol/75µL, n=6, respectivamente), enquanto nas amostras de ratas com cistite, esses valores foram significativamente maiores (955,5±66,3pmol/75µL, n=8, e 926,5±131,7pmol/75µL, n=11, respectivamente). Em ambos os grupos, a NG-monometil-L- arginina causou uma significativa redução na concentração de nitrito/nitrato. Curiosamente, a redução máxima de concentração de nitrito/nitrato causada pela NG-monometil-L- arginina não foi maior que 30% dos valores iniciais. CONCLUSÕES: Esses resultados constituem a primeira demonstração de que as concentrações de nitrito/nitrato no líquido cefalorraquidiano e no interstício da medula espinhal estão elevadas entre 20 e 22 horas após a cistite induzida por ciclofosfamida, e indicando que a cistite está associada a alterações na produção de óxido nítrico, nos segmentos da medula espinhal, nos quais termina a maioria dos aferentes primários da bexiga.
OBJECTIVE: To determine the concentration of nitrate/nitrite in the cerebrospinal fluid and in the dorsal horn interstice of the L6-S1 spinal cord boundary in rats with or without cystitis induced by cyclophosphamide. METHODS: All experiments were conducted using Wistar female rats. A microdialysis probe was implanted in the subarachnoid space or in the spinal cord tissue at the L6-S1 segments (confirmed histologically). Two days later, the microdialysis probe was perfused with artificial cerebrospinal fluid, containing or not NG-monomethyl-L-arginine. Samples were collected every 15 minutes and kept at -20ºC. Nitrite/nitrate concentrations were determined by chemiluminescence. RESULTS: In normal animals, the mean values of nitrite/nitrate concentrations in the first microdialysate sample of the cerebrospinal fluid and of the spinal cord interstice were similar (482.5±90.2pmol/75µL, n=20, and 505.7±11.5pmol/75µL, n=6, respectively), whereas, in the samples from rats with cystitis, these values were significantly greater (955.5±66.3pmol/75µL, n=8, and 926.5±131.7pmol/75µL, n=11, respectively). In both groups, NG-monomethyl-L- arginine caused a significant reduction in the nitrite/nitrate concentration. Interestingly, the maximal reduction of nitrite/nitrate concentration caused by NG-monomethyl-L- arginine was no greater than 30% of the initial values. CONCLUSIONS: These results constitute the first demonstration that nitrite/nitrate concentrations in the cerebrospinal fluid and spinal cord interstice are elevated between 20- and 22 hours after cyclophosphamide-induced cystitis, and indicate that cystitis is associated with changes in the production of nitric oxide in the spinal cord segments, where most primary bladder afferents end.
Subject(s)
Cyclophosphamide , Cystitis/chemically induced , Microdialysis , Nitric Oxide , omega-N-MethylarginineABSTRACT
Objective To evaluate the effects of endothelial nitric oxide synthase (eNOS) gene transfer on intrahepatic vascular resistance (IHVR) and portal venous pressure (PVP) in cirrhotic rats. Methods (1) 5 days after eNOS gene transfer, the in situ liver perfusion system (ISLP) was prepared and in different groups of controls and eNOS treated rats, the followings were analyzed: portal perfusion pressure (PP) dose-response curve to norepinephrine (NE); the effects on PP caused by specific nitric oxide synthase (NOS) inhibitor N-monomethyl-L- arginine (L-NMMA) or the nitric oxide (NO) synthesis substrate L-arginine (L-Arg). (2) The experiment of perfusion via portal vein in vivo was performed and the effects of L-NMMA on the PVP was observed. Results (1) In ISLP model, after L-NMMA was added into the perfusate of the control rats, PP dose-respose to NE increased remarkably and the peak of PP increased to (26.7?0.9) mm?Hg. The increased PP response to NE caused by L-NMMA was offsetted by L-Arg and the peak of PP decreased to (23.2?0.9) mm?Hg. In eNOS treated rats, PP response to NE was significantly lower than that in controls (P