摘要
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 study the effect of nitric oxide (NO) on hepatocyte injury pathway during hepatitis gravis and its possible mechanisms.Methods 24 female BALB/C murine were divided into 4 groups randomly: normal group, model group, NO donor group and NOS inhibitor group. The model of the murine acute hepatitis gravis was established. All animlas were killed 6 h after administration. Serum ALT and TBIL were measured routinely, and the liver tissue was stained with HE. Griess method was applied to determine serum NO 2 - (metabolite of NO). Apoptosis and apoptostic rate of hepatocytes were detected by using TUNNEL in situ assay and FCM, respectively. Results Large amount of NO was produced in model group, accompanied by the increased serum ALT and TBIL. The pathological changes of the injury in liver tissue was evident. When NO intervening factors was adopted, the level of serum NO 2 - in the NO donor group was increased, accompanied by the increased ALT and deterioration of pathological changes in liver tissue; However, the level of serum NO 2 - in the NO inhibitor group was reduced, accompanied by the lowering of ALT and certain alleviation of injury in liver tissue. The apoptotic cells were not detectable in normal liver by TUNNEL and lower than 3 % by FCM. Apoptotic hepatocytes existed in model group with the rate of 20.63 %. The rate of apoptosis in NO donor group was increased to 29.8 %, while that in NO inhibitor group decreased to 10.15 %, and the differences among these groups were obvious. Conclusion NO acts as an apparently injury function on hepatocytes. The pathway of NO -mediated injury is shown as both causing hepatocyte apoptosis and necrosis, which are both involved in the pathogenesis of acute hepatitis gravis. NOS inhibitor can improve the NO -mediated hepatocyte injury to some extent.
出处
《华中科技大学学报（医学版）》
CAS
CSCD
北大核心
2003年第3期269-272,F003,共5页
Acta Medicinae Universitatis Scientiae et Technologiae Huazhong
基金
国家自然科学基金资助项目 (No.392 70 314 )
