摘要
目的:探讨钙通道拮抗剂—维拉帕米(verapamil,Ver)对体外培养的人视网膜色素上皮(humanretinalpigmentepithelium,hRPE)细胞凋亡诱导作用及凋亡过程中细胞内钙浓度[Ca2+]i的变化。方法:应用80mg/L的Ver作用体外培养hRPE细胞12,24及48h,采用吖叮橙(AO)荧光染色法、透射电镜、流式细胞术观察hRPE细胞凋亡;Fluo-3/AM负载技术,观察凋亡过程中细胞[Ca2+]i的变化。结果:一定浓度Ver可诱导hRPE细胞凋亡,荧光显微镜及电镜观察hRPE细胞具有早期凋亡特征:核荧光呈黄绿色,电镜下核染色质浓染、边集。流式细胞术显示,Ver作用后的hRPE细胞凋亡百分率增加(F=12.3415,P<0.05)。Ver作用的hRPE细胞内钙浓度([Ca2+]i)呈下降趋势(F=23.607,P<0.01)。结论Ver能诱导体外培养的hRPE细胞凋亡,细胞内[Ca2+]i浓度的变化可能是诱导hRPE凋亡的机制之一。
AIM： To explore the effect of calcium channel antagonist, verapamil on inducing hRPE apoptosis and the changes of intracellular free [Ca^2＋]i.
METHODS： Cultured hRPE cells were added 80mg/L verapamil for 12, 24 and 48h. AO fluorescence staining, transmission electron microscopy and flow cytometer were used to determine the effect of verapamil on hRPE cells. Fluo-3/AM load technique was used to observed the changes of hRPE intracellular free [Ca^2＋]i.
RESULTS： Verapamil can induce apoptosis of hRPE. Apoptosis features were observed by AO staining with fluorescence microscope： nucleolus fluorescence was kelly. Under transmission electron microscope, nucleolus chromatin concentrated and dyed black. Flow cytometer displayed the apoptosis rate increased （F =12.3415, P 〈 0.05）. hRPE cells intracellular free [Ca^2＋]i decreased gradually （F =23.607, P〈0.01）.
CONCLUSION： Verapamil may induce the apoptosis of cultured hRPE in vitro. The change of [Ca^2＋]i may be the one of important mechanisms of apoptosis of hRPE cells.
出处
《国际眼科杂志》
CAS
2006年第2期324-327,共4页
International Eye Science
基金
中国国家自然科学基金资助(No.30471865)~~
