白介素-1β对大鼠肝切除后剩余肝细胞一氧化氮产生和ATP合成的影响

Effects of Interleukin-1β on Nitric Oxide Production and ATP Synthesis in Rat Surplus Hepatocytes after Partial Hepatectomy

目的探讨大鼠肝部分切除后残存肝分离细胞在白介素-1β(IL-1β)作用下一氧化氮(NO)产生和肝细胞能量代谢的改变。方法将大鼠分为肝部分切除组(PH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IL-1β等细胞因子处理培养肝细胞;应用Griess reagent法检测PH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PH组肝细胞NO产生量约是对照组的2倍。IL-1β能降低两组肝细胞的ATP含量和KBR,PH组降低程度大于对照组。加入L-精氨酸,PH组肝细胞NO的产生增加,ATP水平和KBR降低。一氧化氮合酶抑制剂NG-甲基-L-精氨酸(L-NMMA)可以抑制NO产生,并使降低的肝细胞ATP含量和KBR得以恢复。结论肝部分切除后,在IL-1β作用下的NO产生增加能够抑制ATP合成,促使肝细胞线粒体功能发生障碍。

Objective To detect the nitric oxide （NO） production and energy metabolism of the interleukin （IL）-1β-treated residual hepatocytes from rats after partial hepatectomy. Methods Forty rats were equally divided into partial hepatectomies （PH） group and control group. In the control group the rats were otherwise matched and underwent sham surgeries. The residual hepatocytes were separated by the collagenase perfusion method. The hepatocytes were cultured with cytokines such as IL-1β. The production of NO in the two groups were measured with Griess reagent method, the production of inducible nitric oxide synthase （ iNOS） protein detected with Western blot, the content of the nucleotide in the hepatocytes detected with high-performance liquid chromatography, and the content of the ketone body in the hepatocytes of the two groups determined with the enzymatic method. Afterwards the ketone body ratio （acetoacetate/β-hydroxy butyrate, KBR） was calculated. Results The production of NO in the PH group was twice as much as that in the Sham group. IL-1β decreased the content of ATP and the KBR in the hepatocytes of both groups, and the decrease magnitude in the PH group was significantly larger than that in the Sham group. After the injection of L-arginine, the production of NO in the hepatocytes in the PH group increased, and the level of ATP and KBR decreased. N^G-methyl-L-arginine （ L-NMMA）, the inhibitor of NO synthase, inhibited the production of NO and reversed the decrease of ATP and KBR. Conclusion After partial hepatectomy, increased NO production in the hepatocytes after the treatment of interleukin-1β may disturb the function of mitochondria by inhibiting the synthesis of ATP.