肾上腺素诱导人视网膜色素上皮细胞凋亡的实验研究

Experimental Study on Apoptosis of Culture Human Retinal Pigment Epithelium Induced by Epinephrine

目的 观察肾上腺素对体外培养人视网膜色素上皮 (RPE)细胞凋亡的影响 ,探讨细胞凋亡与中心性浆液性脉络膜视网膜病变的关系。方法 通过流式细胞技术 (FCM )、光镜、TUNEL细胞凋亡原位末端标记法检测肾上腺素和 β 受体阻滞剂普萘洛尔对体外培养的人RPE细胞凋亡的作用。 结果 人RPE细胞对低浓度的肾上腺素有一定耐受作用 ,10 pg/ml～ 10 4pg/ml肾上腺素组无明显凋亡的发生 ;10 5pg/ml～ 10 8pg/ml肾上腺素诱导RPE细胞 72h可以检测到凋亡峰 ,与对照组比较有显著性差异 (P <0 0 1) ;加入 10 3 pg/ml普萘洛尔可以提高RPE细胞对肾上腺素毒性作用的耐受 ,未检见RPE细胞凋亡的发生。TUNEL法光镜下观察到细胞核被蓝色染料标记的凋亡细胞。结论 肾上腺素可以诱导培养的人RPE细胞凋亡 ,这可能是中心性浆液性脉络膜视网膜病变早期病理改变的机制之一。

Objective To observe the effect of epinephrine on the apoptosis of in vitro cultured human retinal pigment epithelial (RPE) cells and investigate the relationship between apoptosis and central serous chorioretinopathy (CSC). Methods The RPE cells were grown to confluence and incubated for 72 h in cultured medium containing epinephrine (10 pg/ml to 10 8 pg/ml) and 10 3 pg/ml propranolol. The changes of RPE cell apoptosis were evaluated by flow cytometry (FCM), light microscopy and in situ terminal deoxynucleotidyl transferase mediated dUTP nick end labeling (TUNEL). Results Exposure to lower concentrations of epinephrine (10 pg/ml to 10 4 pg/ml) could not result in RPE cell apoptosis.10 5 pg/ml-10 8 pg/ml epinephrine could induce apoptosis in a dose dependent manner. Propranolol (10 3 pg/ml) could inhibit epinephrine induced apoptosis. The RPE cells were resistant to epinephrine toxicity even coexposure to 10 8 pg/ml epinephrine in the presence of propranolol. The apoptotic cells marked with blue under the light microscopy were found by TUNEL method. Conclusion The RPE cells may undergo apoptosis following exposure to epinephrine, which suggests a possible pathologic mechanism in the early development of CSC.