维拉帕米对培养的人视网膜色素上皮细胞增殖和DNA合成的影响

目的:观察钙通道阻滞剂维拉帕米(verapamil)对培养的人视网膜色素上皮细胞(human retinalpig-ment epithelium,hRPE)增殖和DNA合成的影响。方法:含不同浓度维拉帕米的条件培养液作用于培养的hRPE细胞后,利用细胞计数和MTT比色试验观察维拉帕米对培养的hRPE细胞的作用,同时采用氚标胸腺嘧啶核苷(3H-thymidine,3H-TdR)掺入试验测定维拉帕米对培养的hRPE细胞DNA合成的影响。结果:维拉帕米在实验所设的浓度范围均能抑制培养的hRPE细胞的增殖和DNA合成,在浓度为10,50,100,200μm ol/L时其细胞增殖抑制率分别为:26.72%,37.25%,58.21%,79.54%(P<0.01);cpm值分别为对照组的0.81,0.61,0.52,0.44倍(P<0.01)。结论:维拉帕米对培养的hRPE细胞增殖和DNA合成具有抑制作用。

Effects on Telomerase Activity and Associated-Protein of hRPE Cells by TGF-β_1

Purpose: To investigate the effects on telomerase activity, human telomerase reversetranscriptase (hTERT) gene and TEP1 mRNA of retinal pigment epithelial (RPE) cells weretreated by TGF-β1 of different concentration.Methods: The cultured human RPE cells were treated with TGF-β1 at differentconcentration(0 ng/ml, 0. 01 ng/ml, 0.1 ng/ml, 1 ng/ml, 10 ng/ml) for 24 h, thentelomerase activity was detected by telomerase repeat amplification protocol(TRAP). Theexpression of hTERT mRNA and TEP-1mRNA were detected by reverse transcriptionpolymerase chain reaction(RT-PCR).Results: TRAP and RT-PCR showed when the concentration of TGF-β1 was graduallyincreased, telomerase activity and the expression of hTERTmRNA were gradually reduced,TEP1mRNA showed no apparent differential expression.Conclusion: TGF-β1 can down-regulate telomerase activity and the expression of hTERTmRNA, but no effection on TEP-1mRNA, hTERTmRNA expression was in accordance withtelomerase activity in hRPE cells, hTERT gene plays a crucial role in the expression oftelomerase activity, while TEP1 plays a much smaller role.

人视网膜色素上皮细胞生长特性及可见光对其影响

研究体外培养的人视网膜色素上皮细胞的生长特性及可见光对其影响。用倒置相差显微镜和透射电镜观察人视网膜色素上皮(hRPE)细胞形态和超微结构,用计数法和噻唑蓝(MTT)法绘制生长曲线。从光照时间、光照时有无血清、光照后是否换液、光照后培养时间这4方面考查可见光对hRPE细胞的损伤情况。实验结果表明,体外培养的hRPE细胞为多角形,有极性,细胞器正常,生长旺盛。(8423±359)lx的可见光光照2h就能对hRPE细胞造成显著损伤。光照时间为5h,光照时培养液中含10%胎牛血清,光照后不换培养液,光照后培养时间为24h,此条件下hRPE损伤率为58%。体外培养的hRPE细胞具有正常的形态结构和生长规律。可见光可对hRPE细胞造成损伤,并且损伤程度有时间积累效应。因此,在日常生活中应尽可能避免阳光直射,减少光照时间,以降低视网膜光损伤发生率,保护眼睛。