铅对体外培养HK-2细胞毒性及碘化钾的拮抗作用  

The toxicity of lead to HK-2 and the antagonistic effects of KI

作　　者：周桂凤[1] 刘栋[1] 乔玉峰[1] 彭佑铭[1] 杨波[1] 蒋云生[2] 

机构地区：[1]中南大学湘雅二医院肾内科,湖南长沙410011 [2]湖南师范大学医学院

出　　处：《毒理学杂志》2008年第4期283-286,共4页Journal of Toxicology

基　　金：湖南省自然科学基金资助项目(05JJ40115)

摘　　要：目的研究醋酸铅对人肾小管上皮细胞系HK-2细胞毒性剂量-反应关系及碘化钾(KI)的拮抗作用。方法体外培养HK-2细胞分为不同剂量染铅组:分别加入浓度为0,2.5,5,10,20和40μmol/L的醋酸铅;KI干预组:加入终浓度为40 mg/L的KI、37℃孵育30 min后加入上述不同剂量的醋酸铅。染毒72 h后,利用噻唑蓝法(MTT)测定各组细胞的存活率;用相差显微镜观察细胞形态改变;Annexin-V/PI双染法检测细胞凋亡并用Hoechst33258染色、荧光显微镜观察细胞凋亡形态。结果细胞存活率与醋酸铅浓度存在剂量-反应关系(r=-0.878,P<0.01),其IC50为15.68μmol/L;低剂量铅组部分细胞形态拉长呈长梭形,随着铅剂量增加,细胞变小、变圆,直至无完整细胞形态、细胞成团,脱壁漂浮;与对照组(0μmol/L铅)比较,各种剂量铅染毒组HK-2细胞凋亡率明显增加(P<0.05),凋亡细胞核染色加深呈高亮蓝色、模糊、核明显固缩和碎裂;各剂量铅组加40 mg/L的KI,可使细胞存活率增加,IC50上升了3.33倍,细胞凋亡率下降,细胞形态改变明显改善。结论醋酸铅对HK-2细胞毒性存在明显剂量-反应关系,KI可部分拮抗铅的细胞毒性。Objective To investigate the dose-response relationship of cytotoxic effects of lead acetate（Pb） on human proximal tubular epithelial cell line （HK-2） and the antagonistic effects of potassium iodide（ KI）. Methods HK-2 cells were divided into two groups. （1）treated with Pb （0,2.5,5,10,20,40 μmol/L）. （2）pretreated with KI of final concentration 40 mg/L. The cell viability was measured with MTT assay and morphological observation. The apoptosis was monitored by using flow cytometry and Hoechst 33258 staining. Results The viability of HK-2 cells was negatively correlated with dosages of Pb （ r = - 0. 878, P 〈 0.01 ） after exposure of Pb for 72 h. The concentration of Pb that inhibited 50% （IC50） of HK-2 cells at 72 h was 15.68 μmol/L. HK-2 cells cultured in high dosages of Pb showed obvious morphologic changes, including elongated, contracted and rounded even agglomerated, apoptosis ratio was elevated, chromatin was condensed and cell nuclei exhibited the dense fluorescence in every dosage of Pb. Compared with same dosages of Pb groups, after pretreatment with KI, cell viability was increased, IC50 being enhanced 3.33 times. Morphologic changes were significantly improved and apoptosis ratio was lowered. Conehlsions There was a dramatically dose-response relationship between dosages of Pb and the toxic effects on HK-2 cells, while 40 mg/L KI could protect Pb-induced cytotoxicity in part.

关 键 词：铅 碘化钾 HK-2细胞 细胞毒性 凋亡 

分 类 号：R99[医药卫生—毒理学]