促红细胞生成素对马兜铃酸致肾小管上皮细胞凋亡的影响  

Effect of erythropoietin on renal tubular epithelial cells stimulated by aristolochic acid

作　　者：王巍巍[1] 张金元[2] 

机构地区：[1]南京军区南京总医院博士后科研工作站,南京210002 [2]解放军第四五五医院南京军区肾脏病专科中心

出　　处：《肾脏病与透析肾移植杂志》2006年第6期530-534,共5页Chinese Journal of Nephrology,Dialysis & Transplantation

基　　金：上海市科学技术发展基金资助项目(054109)

摘　　要：目的:探讨促红细胞生成素(EPO)对马兜铃酸(AA)所刺激的肾小管上皮细胞结构损伤的影响。方法:以不同浓度AA(10、20、40μg/ml)刺激LLC-PK1细胞株,同时培养体系中加入不同浓度的EPO(5、10、20U/ml),另设对照组。各组细胞培养24h后,透射电镜观察细胞的超微结构,TUNEL法原位检测细胞凋亡情况,流式细胞仪检测凋亡细胞的比例。结果:电镜显示:与AA10μg/ml组相比,AA10μg/ml+EPO20U/ml组有少数细胞线粒体肿胀或呈早期凋亡改变(异染色质),大部分细胞结构基本正常;AA10μg/ml+EPO10U/ml组与前者相似,有少数细胞出现凋亡。AA20μg/ml和AA40μg/ml刺激的细胞经不同浓度EPO干预后,细胞的损伤无明显变化。TUNEL结果:经AA10μg/ml、20μg/ml刺激后,核染色阳性的细胞百分比与对照组比较明显增加(P<0·05)。与AA10μg/ml组比较,EPO10U/ml和EPO20U/ml可明显降低阳性细胞的百分比(P<0·05)。流式细胞仪显示:经AA10μg/ml、20μg/ml刺激24h后,细胞凋亡的比例明显增加(P<0·05),AA20μg/ml组凋亡和坏死的细胞比例高于AA10μg/ml组。与AA10μg/ml组比较,EPO10U/ml和EPO20U/ml干预后细胞凋亡比例有所下降,以EPO20U/ml作用显著(P<0·05)。AA20μg/ml组细胞凋亡的比例与EPO干预后的各组比较无显著差异。结论:EPO通过抑制细胞的凋亡从而对AA所刺激的肾小管上皮细胞结构的损伤产生保护作用,但对细胞的坏死改变无明显影响。Objective.. To investigate the effect of erythmpeietin（EPO） on renal tubular epithelial cells stimulated by aristolechic acid（AA）. Methodology：LLC-PK1 cells were stimulated by different concentrations of AA（ 10,20,40 μg/ml ）, and EPO （5,10,20 U/ml）were added as prophylaxis. The media of control contained neither of the agents. Electron microscopy was performed to observe the ultrastructural changes of LLC-PK1 cells. In situ cells, the apoptosis were detected by TUNEL and the proportions of apoptosis were assessed by flow cytometry after 24 hours＇ incubation. Results： The groups of AA10 ＋ EPO 10 U/ml and AA10 ＋ EPO 20 U/ml had few cells appeared chondriosome swelling, heterochromatin or individual apoptosis compared with AA10 μg/ml, while the most cells＇ structure was generally normal. But EPO had no apparent effect on cells stimulated by AA20 μgml and AA40 μgml. The apoptosis proportions increased remark- ably after stimulated by AA10 μg/ml and 20 μg/ml for 24 hours assessed by TUNEL and flow cytometry （ P 〈 0. 05 ）. EPO 10 U/ml and 20 U/ml reduced the proportions of apoptosis induced by AA10 μg/ml（ P 〈 0. 05 ）. Conclusion： EPO can protect LLC-PK1 cells from injury by inhibiting the apoptosis cells stimulated by aristolochic acid.

关 键 词：促红细胞生成素 马兜铃酸 肾小管上皮细胞 

分 类 号：R692[医药卫生—泌尿科学]