CAJ | 학술논문

目的探讨溴苯腈对离体小鼠肝组织线粒体膜电位及呼吸控制率(RCR)的影响及N-乙酰半胱氨酸(NAC)的干预作用.方法采用差速离心法分离提取ICR雄性小鼠肝组织线粒体,随机分成正常对照组、溴苯腈染毒组、NAC保护组,用氧电极法测线粒体Ⅲ态呼吸速率(S3)、Ⅳ态呼吸速率(S4)、RCR;用阳离子荧光羰花青染色剂(JC-1)染色,共聚焦荧光显微镜观察线粒体膜电位变化.结果与正常对照组比较,溴苯腈染毒组的S3为(0.031±0.008)nanoatoms oxygen·mg-1·min-1、RCR为1.820±0.181,NAC保护组的RCR为4.253±0.210,均明显降低,溴苯腈染毒组的S4(0.017±0.004)nanoatoms oxygen·mg-1·min-1明显升高,差异均有统计学意义(P<0.01);与溴苯腈染毒组比较,NAC保护组的S3为(0.046±0.005)nanoatoms oxygen/mg-1·min-1]、RCR(4.253±0.210)明显升高,S4[(0.011±0.001)nanoatoms oxygen·mg-1·min-1明显降低,差异均有统计学意义(P<0.01).共聚焦荧光显微镜观察,溴苯腈染毒组线粒体红色荧光明显减弱甚至消失,NAC保护组线粒体红色荧光较溴苯腈染毒组增强,但较正常对照组弱,3组间差别明显.结论离体小鼠肝组织线粒体经溴苯腈染毒后,线粒体RCR与膜电位明显下降;经NAC预处理后,可有效提高溴苯腈染毒线粒体的RCR与膜电位,对线粒体有较好的保护作用.
목적 탐토추분정대리체소서간조직선립체막전위급호흡공제솔(RCR)적영향급N-을선반광안산(NAC)적간예작용.방법 채용차속리심법분리제취ICR웅성소서간조직선립체,수궤분성정상대조조、추분정염독조、NAC보호조,용양전겁법측선립체Ⅲ태호흡속솔(S3)、Ⅳ태호흡속솔(S4)、RCR;용양리자형광탄화청염색제(JC-1)염색,공취초형광현미경관찰선립체막전위변화.결과 여정상대조조비교,추분정염독조적S3위(0.031±0.008)nanoatoms oxygen·mg-1·min-1、RCR위1.820±0.181,NAC보호조적RCR위4.253±0.210,균명현강저,추분정염독조적S4(0.017±0.004)nanoatoms oxygen·mg-1·min-1명현승고,차이균유통계학의의(P<0.01);여추분정염독조비교,NAC보호조적S3위(0.046±0.005)nanoatoms oxygen/mg-1·min-1]、RCR(4.253±0.210)명현승고,S4[(0.011±0.001)nanoatoms oxygen·mg-1·min-1명현강저,차이균유통계학의의(P<0.01).공취초형광현미경관찰,추분정염독조선립체홍색형광명현감약심지소실,NAC보호조선립체홍색형광교추분정염독조증강,단교정상대조조약,3조간차별명현.결론 리체소서간조직선립체경추분정염독후,선립체RCR여막전위명현하강;경NAC예처리후,가유효제고추분정염독선립체적RCR여막전위,대선립체유교호적보호작용.
Objective To demonstrate the effect of bromoxynil on membrane potential and respiratory control rate (RCR) in isolate mitochondria from mice liver tissue in vim and the intervention of NAC. Methods The mitochondrial was randomized to control group, bromoxynil-poisoned group and NAC-prospected group. S3,S4 and RCR of the mitochondria in each sample was detected by the method of oxygen electrode.Each sample was stained by JC-1 and the changes of membrane potential of mitochondria were observed under fluorescence microscope. Results The S3 [(0.031±0.008)nanoatoms oxygen·mg-1·min-1)],RCR (1.820±0.181) of bromoxynil-poisoned group and RCR (4.253±0.210) of NAC-prespected group were significantly lower than those of control group (P<0.01);the S4[(0.017±0.004) nanoatoms oxygen·mg-1·min-1) of NAC-prospected group was significantly higher than control group (P<0.01).The S3[(0.046±0.005) nanoatoms oxygen·mg-1·min-1)] and RCR of NAC-prospected group were significantly higher than group B (P<0.01),S4 [(0.011±0.001) nanoatoms oxygen·mg-1·min-1)] of NAC-prospected group was significantly lower than bromoxynil-poisoned group (P<0.01). Observaion under fluorescence microscope: the red fluorescence of mitochondria was dim or disappearred in bromoxynil-poisoned group while brightened in NAC-prospected group but still dimmer than control group Conclusion In vitro, the mitochondrial RCR and the mitochondrial membrane potential are decreased after the mitochondria is incubated with bromoxynil, and NAC could improve it.