CAJ | 학술논문

目的】研究铅对肾上腺皮质细胞氧化应激和线粒体功能的影响，为了解其肾上腺皮质毒作用机制提供依据。【方法】原代分离培养豚鼠肾上腺皮质细胞，以0、6.25、12.5、25、50、100 μmol/L醋酸铅(PbAc)处理细胞，观察PbAc诱导肾上腺皮质细胞活性氧(ROS)产生和线粒体损伤作用。ROS检测采用荧光分光光度法，线粒体膜电位(MMP)和细胞存活状态采用Rh123和PI双标记、流式细胞术检测，细胞ATP水平采用化学发光法测定。【结果】 PbAc染毒后肾上腺皮质细胞ROS形成水平随剂量增加而增加,具有剂量效应关系[y=4.16+10.21×1g(x+1)，P<0.01，R2=0.64 1］；线粒体膜电位呈剂量依赖性降低，Rh123的平均荧光强度(MFI)在6.25～100 μmol/L各剂量组依次为1.01,0.94,0.96,0.95和0.91，与对照组(1.35)比较具有显著性差异(P<0 .01)；染毒后细胞死亡率轻度增加，与对照组(1.02%)比较，50 μmol/L和100 μmol/L的 PbAc组分别为3.16%和3.40%，差异具有显著性(P<0.05)；ATP水平降低与PbAc剂量之间存在剂量效应关系[y=212965.7-51592.5×1g(x+1)，P<0.01,R2= 0.568］，剂量和时间对ATP水平的影响呈协同抑制作用(P<0.01)。【结论】线粒体氧化应激介导肾上腺皮质细胞毒性可能是PbAc毒作用机制之一，线粒体损伤是铅致肾上腺皮质毒作用的早期细胞和分子事件。
목적】 연구연대신상선피질세포양화응격화선립체공능적 영향，위료해기신상선피질독작용궤제제공의거。【방법】 원대분리배양돈서신상선피질 세포，이0、6.25、12.5、25、50、100 μmol/L작산연(PbAc)처리세포，관찰PbAc유도신 상선피질세포활성양(ROS)산생화선립체손상작용。ROS검측채용형광분광광도법，선립체막 전위(MMP)화세포존활상태채용Rh123화PI쌍표기、류식세포술검측，세포ATP수평채용화학 발광법측정。【결과】 PbAc염독후신상선피질세포ROS형성수평수제량증가이증가,구유제 량효응관계[y=4.16+10.21×1g(x+1)，P<0.01，R2=0.64 1］；선 립체막전위정제량의뢰성강저，Rh123적평균형광강도(MFI)재6.25～100 μmol/L각제량조 의차위1.01,0.94,0.96,0.95화0.91，여대조조(1.35)비교구유현저성차이(P<0 .01)；염독후세포사망솔경도증가，여대조조(1.02%)비교，50 μmol/L화100 μmol/L적 PbAc조분별위3.16%화3.40%，차이구유현저성(P<0.05)；ATP수평강저여PbAc제량지 간존재제량효응관계[y=212965.7-51592.5×1g(x+1)，P<0.01,R2= 0.568］，제량화시간대ATP수평적영향정협동억제작용(P<0.01)。【결론】 선립체 양화응격개도신상선피질세포독성가능시PbAc독작용궤제지일，선립체손상시연치신상선피 질독작용적조기세포화분자사건。
【Objective】 To study the effect of lead acetate on m itochondria oxidative stress and functional alteration of adrenocortical cells. 【Methods】 Male guinea pig fasciculata-glomerulosa (FG) cells were dispersed and primarily cultured as the testing system. The changes in the formation of re active oxygen species (ROS) and the mitochondria damage were observed when the c ells were incubated with lead acetate at different dosages of 0, 6.25, 12.5, 2 5, 50 and 100 μmol/L. The indices and their corresponding detecting methods inc luded: ① ROS (fluorescence spectrometry); ② Mitochondria membrane potential (M MP) and dead cells rate ［flow cytometry with dual-labeling of rhodamine 123(Rh 123) and propidium iodide(PI)］; ③ ATP level (chemoluminescence assay). 【Resul ts】 The formation of ROS in adrenocortical cells was increased in dose-depen dent manner after lead acetate incubation ［y=4.16+10.21×1g(x +1)，P<0.01，R2=0.641］；MMP decreased as the dose of PbAc increased . The mean fluorescent intensity(MFI) of Rh123 dropped from 1.01 to 0.91 as th e dosage increased from 6.25 μmol/L to 100 μmol/L, which were significantly lower than that of the control (1.35, P<0.01)； Dead cells rate appeared to be slightly increased. Dead cells rates in the grou ps of 50 μmol/L and 100 μm ol/L PbAc were 3.16% and 3.40% respectively when compared with the control (1 .02%, P<0.05)；ATP levels tended to decrease, resulting from the synergist ic inhibition by the dose and incubation period of PbAc. The regression analysis showed that dose-effect relationship existed between the decreased ATP and the increased dosage of PbAc ［y=212965.7-51592.5×1g(x+1)，R 2=0.568，P <0.01］. 【Conclusion】 This study demonstrated that cytotoxicity of adrenoco rtical cells induced by lead acetate is probably attributed to mitochondria oxid ative stress. Mitochondria damage might be the early cellular and molecular even t in toxic effect of lead on adrenal cortex.