金雀异黄素诱导培养人视网膜色素上皮细胞凋亡

Genistein induces the apoptosis of cultured human retinal pigment epithelial cells

目的 :探讨金雀异黄素对培养人视网膜色素上皮细胞 (hRPE)的促凋亡作用 .方法 :将不同浓度的金雀异黄素作用于培养人视网膜色素上皮细胞 ,通过TUNEL染色和光、电镜的形态学观察 ,观察金雀异黄素对培养hRPE凋亡的诱导作用 .结果 :不同浓度的金雀异黄素可以诱导RPE凋亡 ,5 0mg·L-1 金雀异黄素作用 2 4h即有TUNEL阳性的凋亡细胞出现 ,作用时间在 2 4 ,4 8和 72h ,其凋亡细胞百分数的中位数分别为 7.6 % ,9.8%和 1 3.7% ;当金雀异黄素浓度为 75和1 0 0mg·L-1 时 ,以上 3个时间点凋亡细胞百分数的中位数分别为 1 0 .3% ,1 6 .4 %和 2 3.4 % ;1 5 .4 % ,2 1 .2 %和 35 .8% .透射电镜观察 ,5 0mg·L-1 浓度作用于细胞 ,胞核内染色体出现边集 ,表现凋亡的早期征象 ;75mg·L-1 作用呈现典型的凋亡特征 ;1 0 0mg·L-1 作用于细胞 ,除凋亡细胞外 ,部分细胞出现胞质的进行性溶解 ,有胞膜破坏现象 ,坏死细胞比例增多 .结论 :金雀异黄素可诱导RPE凋亡的发生 ,并有剂量和时间效应关系 .1 0 0mg·L-1

AIM: To investigate the induction of genistein on the apoptosis of the cultured human retinal pigment epithelial (hRPE) cells. METHODS: Different concentration of genistein was administrated to the culture of the hRPE cells. Cell apoptosis were detected by TUNEL methods. The morphologic changes of cells were observed by light microscopy and transmission electron microscopy. RESULTS : TUNEL positive RPE cells were detected under treatment of 50 mg·L -1 genistein for 24 h. At 24, 48, and 72 h, the median of percentage of TUNEL positive cells treated with 50 mg·L -1 genistein was 7.6%, 9.8%, and 13.7% respectively, 10.3%, 16.4%, and 23.4% respectively when treated with 75 mg·L -1 genistein, and 15.4%, 21.2%, and 35.8% respectively when treated with 100 mg· L -1 genistein. After 50 mg·L -1 genistein treatment, hRPE cells exhibited earlier stage of apoptosis presentation. While after 75 mg·L -1 genistein treatment, RPE cells showed condensation of nuclear chromatin and dense apoptosis bodies, which was identified typical apoptosis characterization. When treated with 100 mg·L -1 genistein,besides apoptosis manifestation, parts of RPE cells presented progressively lysis of cytoplasm and destruction of cell membrane, the necrosis rate increased. CONCLUSION: Genistein can induce the apoptosis of hRPE cells in a dose and time dependent manner. 100 mg·L -1 genistein can induce RPE cells necrosis.