Nicoletti V G, Caruso A, Tendi E A, Privitera A, Console A, Calabrese V, Spadaro F, Ravagna A, Copani A, Stella A M
Institute of Biochemistry, Faculty of Medicine, University of Catania, Italy.
Biochimie. 1998 Oct;80(10):871-81. doi: 10.1016/s0300-9084(00)88882-3.
In the present study we evaluated the effects of NO synthase (NOS) induction on the regulation of cytochrome c oxidase (CO) and F0F1-ATPase subunit expression in astroglial and mixed cortical cell cultures. In mixed cortical cell cultures, 18 h of treatment with lipopolysaccharide (LPS, 0.1 microgram/mL) plus interferon-gamma (INF-gamma, 10 U/mL) caused an increase of mRNAs for CO-I, F0F1-ATPase 6 and also for iNOS at 20 DIV. The induction of both CO-I and F0F1-ATPase 6 was abolished by the NOS inhibitor N-monomethyl-L-arginine (NMMA) or by the enzymatic scavenger superoxide dismutase/catalase (SOD/CAT). In primary astroglial cell cultures, treatment for 18 h with increasing concentrations of LPS and INF gamma, produced an increase in the amount of mitochondrial encoded CO-I and -II subunits, with no significant modifications of nuclear encoded subunit IV. An increase was also observed at level of transcription for CO-I and -II, and F0F1-ATPase 6 mRNAs. These effects were abolished by addition of NMMA or SOD/CAT. mRNA induction of CO-I was higher in mixed cortical than in astroglial cell cultures while that of F0F1-ATPase 6 was similar in both cell types. These results suggest that the expression of mitochondrial encoded subunits (CO-I, CO-II and F0F1-ATPase 6) is up-regulated in response to oxygen and NO reactive species. The activity of cytochrome c oxidase decreased after LPS/INF gamma treatment in both astroglial and mixed cortical cultures. The activity of ATP synthase was unmodified, while ATP content drastically decreased after LPS/INF gamma treatment, in both astroglial and mixed cortical cultures. The enzymatic activities of catalase and Mn-SOD (mitochondrial) showed a significant increase after LPS/INF gamma treatment, which was abolished by NMMA.
在本研究中，我们评估了一氧化氮合酶（NOS）诱导对星形胶质细胞和混合皮质细胞培养物中细胞色素c氧化酶（CO）和F0F1 - ATP酶亚基表达调控的影响。在混合皮质细胞培养物中，用脂多糖（LPS，0.1微克/毫升）加干扰素 - γ（INF - γ，10单位/毫升）处理18小时，导致在培养20天时CO - I、F0F1 - ATP酶6以及诱导型NOS（iNOS）的mRNA增加。NOS抑制剂N - 单甲基 - L - 精氨酸（NMMA）或酶清除剂超氧化物歧化酶/过氧化氢酶（SOD/CAT）可消除CO - I和F0F1 - ATP酶6的诱导。在原代星形胶质细胞培养物中，用浓度递增的LPS和INFγ处理18小时，使线粒体编码的CO - I和 - II亚基的量增加，而核编码的亚基IV没有明显变化。在CO - I和 - II以及F0F1 - ATP酶6 mRNA的转录水平也观察到增加。添加NMMA或SOD/CAT可消除这些作用。混合皮质细胞培养物中CO - I的mRNA诱导高于星形胶质细胞培养物，而F0F1 - ATP酶6在两种细胞类型中相似。这些结果表明，线粒体编码的亚基（CO - I、CO - II和F0F1 - ATP酶6）的表达响应于氧和NO反应性物质而上调。在星形胶质细胞和混合皮质培养物中，LPS/INFγ处理后细胞色素c氧化酶的活性降低。ATP合酶的活性未改变，而在星形胶质细胞和混合皮质培养物中，LPS/INFγ处理后ATP含量急剧下降。LPS/INFγ处理后，过氧化氢酶和锰超氧化物歧化酶（线粒体）的酶活性显著增加，这被NMMA消除。
