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目的 探讨NO对重型肝炎肝细胞损伤方式的影响及可能机制。方法 雌性BALB/C小鼠 2 4只随机分为正常对照组、模型组、L Arg组 (NO供体干预 )和L NAME组 (NOS抑制剂干预 )。各组动物均于用药后 6h处死。常规测定血清谷丙转氨酶 (ALT)、血清总胆红素 (TBIL)并进行肝组织HE染色。用Griess法测定血清NO代谢产物NO2 -,末端转移酶标记技术原位检测各组肝细胞凋亡状况及采用流式细胞术 (FCM)检测各组肝细胞凋亡率。结果 急性重型肝炎模型组动物体内产生大量NO ,ALT、TBIL上升 ,出现重肝的肝组织病理学改变。运用NO合成干预因素后 ,L Arg组血清NO2 -升高 ,伴随ALT的上升和肝组织病变的加重 ;L NAME组NO2 -水平降低、ALT水平下降且肝组织损伤有一定程度的改善。正常肝细胞生理凋亡率低于 3% ,模型组凋亡率达 2 0 6 3% ,L Arg组凋亡率上升至 2 9 8% ,而L NAME组凋亡率下降到 10 15 % ,组间比较有显著性差异。结论 NO具有明显的肝细胞损伤作用 ,且坏死和凋亡同时存在。NOS抑制剂可在一定程度上减轻NO对肝细胞的损伤 (包括凋亡和坏死 )。
Objective To investigate the effect of NO on the hepatocyte injury in severe hepatitis and its possible mechanism. Methods Twenty four female BALB / C mice were randomly divided into normal control group, model group, L Arg group (NO donor intervention) and L NAME group (NOS inhibitor intervention). All animals were sacrificed at 6h after treatment. Routine determination of serum alanine aminotransferase (ALT), serum total bilirubin (TBIL) and liver tissue HE staining. The serum NO metabolites NO2 - was measured by Griess method. The apoptosis of hepatocytes in each group was detected by end-transferase-labeled technique. The apoptosis rate of hepatocytes in each group was detected by flow cytometry (FCM). Results Acute severe hepatitis model group produced a large number of animals NO, ALT, TBIL increased, the occurrence of severe liver pathology changes. NO synthesis in L Arg group was increased with the increase of ALT and the increase of liver tissue lesion. NO2 - level decreased, ALT level decreased and liver damage improved to some extent in L - NAME group. The normal hepatocyte apoptosis rate was less than 3%, the apoptosis rate of model group was 2036%, the apoptosis rate of L Arg group was increased to 298%, while the apoptosis rate of L NAME group was reduced to 10 15% There were significant differences between groups. Conclusion NO has obvious hepatocellular injury, and necrosis and apoptosis exist simultaneously. NOS inhibitors can reduce the damage of NO to hepatocytes (including apoptosis and necrosis) to a certain extent.