视网膜血管内皮细胞和周细胞在酸性环境中的增生和凋亡  

Investigation of proliferation and apoptosis of retinal vascular endothelial cells and pericytes under acidic extracellular pH

作　　者：朱冬青[1] 许迅[1] 郑志[1] 顾青[1] 

机构地区：[1]上海交通大学附属第一人民医院眼科,200080

出　　处：《眼科研究》2008年第9期663-667,共5页Chinese Ophthalmic Research

摘　　要：目的探索酸中毒在视网膜新生血管生长的细胞机制中的作用。方法原代培养获取牛视网膜血管内皮细胞(BRECs)和周细胞(BRPs);将传代后的细胞暴露在酸性环境中,通过细胞活力、细胞周期和细胞凋亡检测,观察细胞的增生和凋亡状况。结果细胞培养得到纯化的BRECs和BRPs;酸中毒明显抑制BRECs和BRPs的生长(P<0.01);使处于S期的BRECs和BRPs减少(P<0.01),DNA合成受抑制。重度酸中毒抑制BRECs凋亡(P<0.05),引起BRPs凋亡(P<0.01)。结论酸中毒引起BRPs凋亡而抑制BRECs凋亡,提示视网膜酸中毒参与了糖尿病视网膜病变(DR)的新生血管生长过程。Objective Retinal acidosis follows hyperglycemia and hypoxia. The purpose of this study was to explore the role of acidosis in the cellular mechanism of retinal neovascularization. Methods Primary bovine retinal endothelial cells （BRECs） and bovine retinal pericytes （BRPs） were obtained from bovine and were cultured in DMEM ＋ 20% fetal bovine serum. The passaged retinal endothelial cells and pericytes were exposed to various culture condition with pH 7.2,pH 6.8 or pH 6.5. Proliferation rate in normal and acidified cells was determined by MTS,and apoptosis and cycles assay staining were analyzed by Annexin V/FITC and PI staining using flow cytometry, Results Cultured BRECs were identified by anti-Ⅷ factor relavent antigen antibody, showing the brown staining. Cultured bovine retinal pericytes showed α-SMA positive staining, There was statistically significant difference in cultured cell number （ F =432. 910,P =0. 000） ,cell number of S phase （ F = 29. 000,P = 0.001） and cellularapoptosis rate （F=7. 14,P=0.024 ） of BRECs among the pH 7. 2 group,pH6.8 group and pH6.5 group. The growth status of cultured BRPs showed the same pattern （ F = 147. 640,P =0. 000;F =26. 680,P =0, 001 ;F = 11. 680,P = 0. 009,respectively）,indicating acidosis significantly inhibited the proliferation and DNA synthesis of BRECs and BRPs （P 〈 0. 01 ） ,and severe acidosis inhibited the apoptosis of BRECs （ P 〈 0.05 ） but induced the apoptosis of BRPs （ P 〈 0. 01 ）. Conclusion The fact that acidosis inhibits the apoptosis of endothelial cells and induces the apoptosis of perieytes reveals that acidosis may participate in the pathogenesis of diabetic retinopathy.

关 键 词：酸中毒 内皮细胞 周细胞 增生 凋亡 

分 类 号：R774.1[医药卫生—眼科]