氯化镉诱发肾上腺皮质细胞凋亡与蛋白激酶B活力的关系  

Relationship between apoptosis and activity of protein kinase B in adrenocortical cells induced by cadmium chloride

作　　者：赵敏[1] 魏青[2] 卢次勇[2] 杨杏芬[1] 

机构地区：[1]广东省疾病预防控制中心毒理所,广州510300 [2]中山大学公共卫生学院

出　　处：《中华劳动卫生职业病杂志》2008年第3期135-138,共4页Chinese Journal of Industrial Hygiene and Occupational Diseases

基　　金：广东省自然科学基金资助项目（021990）

摘　　要：目的了解氯化镉（CdCl2）诱发肾上腺皮质细胞凋亡的发生与蛋白激酶B（protein kinase B，PKB，又称Akt）活力变化的关系。方法体外分离培养豚鼠肾上腺皮质细胞，以蛋白因子添加素Ⅴ和碘化丙啶联合标记，流式细胞仪检测，观察CdCl2诱发细胞凋亡的特征；采用免疫沉淀-化学发光法测定PKB／Akt活力。结果6．25～100．00μmol／L剂量CdCl2处理肾上腺皮质细胞2h，细胞凋亡发生率随剂量增加而增加，25．00μmol／L及更高剂量组与对照组比较，差异有统计学意义（P〈0．01）；回归分析表明，CdCl2剂量与凋亡发生率呈剂量-效应关系。以50．00μmol／L CdCl2处理细胞，细胞凋亡发生率随时间的延长而增加，CdCl2作用15min～4h，平均凋亡率为5．58％～73．08％，其中1、2、4h时间点与对照相比，差异有统计学意义（P〈0．05）；CdCl2处理时间与凋亡发生率呈时间-效应关系。6．25～200．00μmol／L剂量CdCl2处理30min，显示CdCl2可引起肾上腺皮质细胞PKB／Akt表达降低，在一定剂量范围内呈线性关系。结论CdCl2诱发肾上腺皮质细胞凋亡的同时引起细胞内PKB／Akt表达水平的降低。Objective To study apoptosis induced by cadmium chloride （CdCl2） and the alteration in activity of protein kinase B （PKB/Akt） in adrenocortical cells. Methods Fasciculata-glomerulosa（FG） cells of male guinea pigs were dispersed and primarily cultured in vitro. Features of apoptotic cells were observed using combined labeling with annexin-Ⅴ and propidium iodide（PI） and flow cytometry. The activity of PKB/Akt was determined with immunoprecipitation and the chemiluminescence assay. Results Apoptosis rate of FG cells increased with dose of CdCl2 two hours after treatment with 6.25-100.00 μmol/L of it, with significant difference in the groups treated with 25.00,50.00,100.00 μmol/L of CdCl2, as compared with the control group （P〈0.01）. Regression analysis showed that occurrence of apoptosis was correlated with the dose of CdC12 in a dose-response pattern. In the meanwhile, there were obviously elevated percentages of apoptotic cells as the increase in duration of incubation ranging from 5.58% to 73.08% for incubating cells with 50.00 μmol/L of CdCl2, from 15 minutes to 4 hours. Duration of incubating cells with CdCl2 were correlated with occurrence of apoptosis in a time-effect manner. The gray scales of PKB/Akt was manifested to be decreased as the ascending of CdCl2 dosage from 6.25 to 200.00 μmol/L, with the linear correlation. Conclusion 6.25 to 100.00 μmol/L CdCl2 might elicit apoptosis of adrenocortical cells. Meanwhile, PKB/Akt is decreased.

关 键 词：氯化镉 细胞凋亡 肾上腺皮质 蛋白激酶类 

分 类 号：R114[医药卫生—卫生毒理学] R743.31[医药卫生—公共卫生与预防医学]