摘要
目的了解中毒性肾病中细胞凋亡的发生机制。方法用不同浓度的庆大霉素（０．５、１、２、４、８、１６ｍｍｏｌ／Ｌ），作用于体外培养的猪肾小管上皮细胞株（ＬＬＣＰＫ１），观察有无细胞凋亡发生以及细胞凋亡与细胞内Ｃａ２＋浓度变化的关系。结果庆大霉素能引起体外培养的肾小管上皮细胞发生凋亡，该凋亡分别经光镜、电镜、琼脂糖凝胶电泳对ＤＮＡ片断的测定以及原位细胞死亡检测等方法证实。利用流式细胞仪，定量测定表明，随着庆大霉素浓度的增加（２～１６ｍｍｏｌ／Ｌ），凋亡细胞所占比例也增加（８．８％、１０．２％、１６．１％、１８．３％，Ｐ＜０．０１）。通过激光共聚焦显微镜对细胞内Ｃａ２＋浓度的测定，发现庆大霉素组（４ｍｍｏｌ／Ｌ）平均细胞内Ｃａ２＋浓度明显高于正常对照组（Ｐ＜０．０１）。结论庆大霉素能诱导ＬＬＣＰＫ１细胞凋亡，凋亡的发生与培养液中庆大霉素浓度及作用时间有关，凋亡细胞内Ｃａ２＋浓度比正常组显著增加。
Objective To study the mechanism of apoptosis in gentamycin treated LLC PK1 cell. Methods One group of renal proximal tubular epithelium (RPTE) cell line LLC PK1 was treated with different concentrations of gentamycin(1,2,4,8,16mmol/L) and another group served as control. RPTE cell apoptosis was identified by light microscopy, electronmicroscopy, agaros gel electronphoresis and in situ cell death test. Results Cell membrane blebbing was seen under inversed microscopy. “Ladder” and nuclear margination and condensation were also identified separately. Quantitative analysis was done by flow cytometry(FACS) using propidium iodide (PI) staining. The ratio of apoptosis increased as gentamycin concentration increased(8.8%, 10.2%, 16.1%, 18 3%). Average calcium concentration assembled by fluorescences condensation was meassured by lazer confocus microscopy using Fluo 3/Am staining. Intracellular calcium concentration in RPTE treated with gentamycin increased markedly as sompared with that in normal cells. Conclusion 1. Gentomycin facilitates RPTE apoptosis in a dose dependent manner. 2. Calcium concentration in the apoptotic cells elevated markedly. It is suggested that intracellular calcium variation is propably related to the mechanism of LLC PK1 apoptosis induced by gentamycin.
出处
《中华内科杂志》
CAS
CSCD
北大核心
1998年第10期667-670,共4页
Chinese Journal of Internal Medicine
关键词
肾功能衰竭
上皮样细胞
庆大霉素
细胞凋亡
钙
Apoptosis Kidney failure, acute Epithelial cell Gentamycins LLC PK1 cells
