高糖对人晶状体上皮细胞凋亡的作用

Effect of high glucose with various concentration on apoptosis of human lens epithelial cells

目的：研究不同浓度的高糖（HG）对人晶状体上皮细胞凋亡的作用。方法：用不同浓度的HG诱导人晶状体上皮细胞凋亡,处理组：DMEM＋HG;对照组：DMEM。用四甲基偶氮唑盐（MTT）比色法检测人晶状体上皮细胞的生长活性;原位末端核苷标记（TUNEL）法检测细胞凋亡的凋亡指数（AI）,并在电镜下观察TUNEL染色阳性细胞的形态及分布。结果：HG可抑制人晶状体上皮细胞生长,诱导细胞的凋亡,并呈剂量及时间依赖性,HG诱导组细胞凋亡率明显高于正常组（P〈0.01）。TUNEL染色法在电镜下观察到HG诱导24h后,TUNEL染色阳性细胞核固缩或碎裂呈棕黄色,细胞大小不一;而TUNEL染色阴性细胞着色均匀,大小均一。结论：高糖具有促使人晶状体上皮细胞凋亡形成的作用,尤其是在120～150mmol/L浓度范围内。

AIM. To investigate the effect of high glucose （HG） with various concentration on apoptosis of human lens epithelial cells （hLECs）. METHODS. hLECs were divided into 2 groups, the normal control group （DMEM） and the treated group （DMEM ＋ HG）. The inhibition of the HG with various concentration on activity of hLECs was analyzed by MTT chromatometry. The induction of high glucose on apoptosis of hLECs （Apoptotic Index, AI） was analyzed by the terminal deoxynucleotidy I transferase-mediated dUTP-biotin nick end labeling （TUNEL） method, the morphous and distribution of the TUNEL-positive cells was also observe by electron microscope. RESULTS. The activity of hLECs was inhibited by HG, and the apoptosis of hLECs was induced by HG, with a dose and time dependence effects. TUNEL-positive cells rate in the treated group was higher than that in the normal control group （ P〈 0.01 ）. Twenty-four hours after incubation with the HG, the nucleus of TUNEL-positive cells was to be pyknoic or was broken to pieces with the dark buffy color, while the TUNEL- negative cells was well-distributed with the light brown color. CONCLUSION. Apoptosis of hLECs can be induced by HG, especially with the concentration between 120mM to 150mM.