水蛭渗滤液对凝血酶诱导的人视网膜色素上皮细胞周期的影响

目的 研究水蛭渗滤液对凝血酶诱导的人视网膜色素上皮细胞周期的影响,从而探讨中药水蛭用于防治PVR的作用机制.方法 (1)采用消化培养法,对人视网膜色素上皮细胞进行体外传代培养. (2)在前期实验基础上选择128mg/ml、64mg/ml、32mg/ml、16mg/ml、8mg/ml浓度的水蛭渗滤液,用流式细胞仪测定细胞周期,观察水蛭渗滤液单独或与0.5NIHU/ml凝血酶共同作用于人视网膜色素上皮细胞24h对细胞周期的影响. (3)选择64mg/ml水蛭渗滤液,观察单独或与0.5NIHU/ml凝血酶共同作用于人视网膜色素上皮细胞6、12、24和48h对细胞周期的影响.结果 (1)水蛭渗滤液在所选择浓度范围内具有使人视网膜色素上皮细胞抑制在G0/G1期的趋势,其中最显著的是64mg/ml水蛭渗滤液可以将77.9%的人视网膜色素上皮细胞抑制在G0/G1期,使进入S期的细胞数减少.细胞增殖活性降低. (2)在观察的时限内,64mg/ml水蛭渗滤液随作用时间的延长,能使更多的人视网膜色素上皮细胞抑制在G0/G1期,增殖活性降低.结论 证实了水蛭渗滤液能抑制体外培养的人视网膜色素上皮细胞增殖.其机制可能是作用于细胞周期,使更多的视网膜色素上皮细胞抑制在G0/G1期,进人S期的细胞数减少,细胞增殖活性降低.

The effects of leech percolate on cell dividing cycle of human retinal pigment epithelium cells induced by thrombin.

Objective To investigate the effects of leech percolate on cell division cycle of culturedhuman retinal pigment epithelium (hRPE)cells induced by thrombin.Exploring mechanism of leech percolateon preventing and curing PVR.Methods (1)Human retinal pigment epithelium(hRPE)cells were cultured invitro with digest culture method. (2)The selected concentrations of leech percolate (128mg/ml,64mg/ml,32mg/ml,16mg/ml and 8mg/ml)were based on the previous experiments.With or without 0.5NIHU/mlthrombin, hRPE cells were treated with leech percolate by various concentrations for 24hrs, and the effects ofcell division cycle of leech percolate were studied with Flow Cytometer (FCM). (3)With or without0.5NIHU/ml thrombin, hRPE cells were treated with 64mg/ml leech percolate at different times(6hr, 12hr,24hrand 48hr), and the effects of cell division cycle of leech percolate were treated with FCM.Results (1)Therewas a general tendency that leech percolate of the selected concentrations arrested hRPE cells on G0/G1 phaseand 64mg/mi leech percolate inhibited 77.9% hRPE cells on G0/G1 phase and made hRPE cells on S phaseless especially. (2)During the period of 6hrs and 48hrs the longer the treat time of 64mg/ml leech percolate, themore the RPE cells arrested on G0/G1 phase.Conclusion The results approve leech percolate can inhibitproliferation of human RPE cells cultured in vitro.Its mechanism may be that leech percolate could act on celldivision cycle and arrest RPE cells on G0/G1 phase to make proliferation power of hRPE cells lower.