一氧化氮介导的线粒体损害诱发败血症肝功衰竭的研究  

No-mediated mitochondria damage induce liver function failure in rats sepsis

作　　者：涂巍[1] 曲文志[1] 赵熳[1] 于作夫[1] 金光华[1] 王勤勇[1] 胡松[1] 

机构地区：[1]中国医科大学附属第四医院普外科,沈阳110032

出　　处：《中华实验外科杂志》2008年第5期599-601,共3页Chinese Journal of Experimental Surgery

基　　金：辽宁省教育厅高等学校科学技术研究项目05L565

摘　　要：目的通过检测败血症大鼠分离肝细胞一氧化氮（NO）的产生，探讨NO对肝细胞功能障碍的影响。方法将大鼠分成盲肠结扎穿刺手术组（CLP组）和假手术对照组（Sham组），采用胶原酶灌注法分离剩余肝细胞并进行培养；应用白细胞介素（IL）-1β等细胞因子处理培养肝细胞；应用Griess reagent法检测CLP组和Sham组肝细胞NO的产生量；应用Western blot检测两组诱导型一氧化氮合酶（iNOS）蛋白的产生；应用高效液相色谱法测定两组肝细胞核苷酸含量；应用酶法检测两组肝细胞的酮体含量并计算酮体比率（乙酰乙酸盐／β-羟基丁酸盐，KBR）。结果CLP组肝细胞NO的产生量是对照组的2～3倍。IL-lβ能够降低两组肝细胞的ATP含量和酮体比率，CLP组降低程度大于对照组。加入L-精氨酸（L-Arg），CLP组肝细胞一氧化氮的产生增加，ATP水平和KBR降低。一氧化氮合酶（NOS）抑制剂NG-甲基-L-精氨酸（L-NMMA）可以抑制NO的产生，并使降低的肝细胞ATP含量、KBR得以恢复。结论对败血症的应答导致NO合成的增加可能与败血症大鼠肝功能障碍有关。对NO产生的调节可能会成为预防败血症所致肝功能障碍的有效方法。Objective By the means of detecting the production of Nitric oxide （NO） in the hepatocytes isolated from rats with sepsis we discussed the influence of NO on the dysfunction of the hepatocytes. Methods Firstly, divided the rats into two groups, one group in which the rats went through the process of Cecal ligation and puncture （the CLP group） ,the other matched control group in which the rats went through a sham surgery process （the Sham group） ; Secondly, separated the hepatocytes through the coUagenase perfusion method, and cultivated the very hepatocytes with some cytokines, such as IL-1β. Then, measured the production of NO in the two groups with the Griess reagent method, detected the production of iNOS protein with Western blot method, assayed the content of the nueleotide in the bepatocytes with the high-performance liquid chromatography （HPLC） ,and determined the content of the ketone body in the hepatocytes of the two groups with the enzymatic method,then calculated the ketone body ratio （acetoacetate/ β-hydroxy butyrate,KBR）. Results The production of NO in the CLP group is twice to third as many as that in the Sham group,IL-1β can decrease the content of ATP and the KBR in the bepatocytes of the two groups, and the decrease degree in the CLP group is greater than that in the Sham group. When the L-arginine was injected,the production of NO in the hepatocytes in the CLP group is increased,and the level of ATP and KBR is decreased. NG-methyl-L-arginine （L-NMMA）, the inhibitor of NO synthetase, can inhibit the production of NO, and recuperate the decrease of ATP and KBR. Conclusion Enhanced production of NO primed by the septic response may be associated with hepatic dysfunction in the liver of septic rats. The regulation of the production of NO may be an effective method in the prevention of the hepatic dysfunction in sepsis.

关 键 词：败血症 一氧化氮 肝功能障碍 

分 类 号：R575.3[医药卫生—消化系统]