Interaction of nitric oxide and reactive oxygen on ADP-ribose-polymerase activation DNA damage

目的：通过抑制多聚腺苷二磷酸核糖聚合酶的激活和NO的产生，观察缺血再灌注时细胞内ATP水平、DNA损伤和器官损伤，以揭示缺血再灌注损伤的机制。方法：60min。缺血180min。对雄性Wistar大鼠70%的肝脏进行再灌流。3AB组大鼠在缺血前给予PARS抑制剂3-氨基苯甲酰胺，L-NAME组大鼠在缺血前不给予任何抑制剂L-NAME。对照组大鼠注入生理盐水。假手术组大鼠仅行肝门区切除，无缺血再灌流。观察缺血再灌流肝(IR-肝)和非缺血再灌流肝(NIR-肝)的组织损伤和DNA损伤。用硝基酪氨酸染色评价ONOO-对组织的损伤。测定各组大鼠血清碱性磷酸酶(ALP)、肝内三磷酸腺苷(ATP)和硝基酪氨酸水平。结果：与…相比，L组血清ALT水平最高与对照组和3AB组比较，差异有统计学意义。对照组和L组IR肝组织和DNA损伤较重，3AB组IR肝组织和DNA损伤较轻。4组大鼠肝脏的近红外光谱差异无统计学意义。对照组和3AB组IR-LOVE呈高硝基酪氨酸染色。他们的组织中也含有高浓度的硝基酪氨酸。L组未见任何硝基酪氨酸染色或产物。在IR肝脏中，3AB组肝内三磷酸腺苷水平高于对照组和L组。在近红外肝脏中，尽管对照组和L组的肝内三磷酸腺苷水平低于假手术组，但3AB组的肝内三磷酸腺苷水平与假手术组相同。结论：脑缺血再灌注损伤后多脏器功能衰竭的发生可能与细胞内ATP的消耗和PARs的激活以修复自由基损伤的DNA损伤有关。较少

Purpose : To reveal the mechanism of ischemia reperfusion injury, we evaluate intracellular ATP levels and DNA damage and organ damage in ischemia reperfusion with inhibition of poly(ADP-ribose) polymerase activation and NO production. Methods : 60 min. ischemia and 180 min. reperfusion in the 70% liver of the male Wister rats were performed. The 3AB group rats received PARS inhibitor, 3-aminobenzamide, and the L-NAME group rats received NO inhibitor, L-NAME, before ischemia. The control group rats were infused saline. The sham group rats were treated only dissection of portal area without ischemia reperfusion. Tissue damage and DNA damage was evaluated both in the ischemia reperfused liver (IR-liver) and the non-ischemia reperfused liver (NIR-liver). Nitrotyrosine stain was performed for evaluating tissue damage by ONOO-. Serum ALP levels, intrahepatic ATP levels and Nitrotyrosine levels were also evaluated in each group. Results : The L-NAME group had highest serum ALT levels compare … More d with the control and the 3AB groups. The control and the L- NAME groups had severe tissue and DNA damages in the IR-liver, while the 3AB group had mild tissue and DNA damages in the IR-liver. There were not differences in the NIR-liver among 4 groups. The IR-love in the control and the 3AB groups had high nitrotyrosine staining. They also had high tissue concentrations of nitrotyrosine. The L-NAME group did not have any nitrotyrosine staining or production. In the IR-liver, the 3AB group had higher intrahepatic ATP levels than the control and the L-NAME groups. In the NIR-liver, although the control and the L-NAME groups had lower levels of intrahepatic ATP levels compared with the sham group, the 3AB group had same intrahepatic ATP levels as the sham group. Conclusion : We concluded that multiple organ failure developed in the ischemia reperfusion injury would be due to consumption of intracellular ATP with activation of PARS to repair DNA damage caused by free radicals in ischemia reperfusion. Less