玻璃体液对培养人视网膜微血管内皮细胞和色素上皮细胞增生的影响

目的 探讨玻璃体液对培养的人视网膜微血管内皮细胞(human retinal capillary endothelialcells,HRCECs)和色素上皮(retinal pigmental epithelial cells,RPE)细胞增生的影响.方法 原代培养HRCECs和RPE细胞,鉴定并传至第3代,再分别培养在1:8、1:4、1:2(玻璃体液在总培养液中的体积比)的人玻璃体条件培养液(vitreous-conditioned medium,VCM)中,其中VCM分为有无血清2组,在不同作用时间(24～72h),采用四唑盐(tetrazolium,MTT)比色法检测VCM对人HRCECs和RPE细胞增生的影响.结果 在有血清时.与对照组比较,1:4、1:2的VCM在培养的各时间段对HRCECs的抑增生作用差异有显著性意义(P<0.05),而对RPE细胞和混合细胞(HRCECs和RPE混合1:1)的促增生作用差异有统计学意义(P<0.01).在无血清时与对照组比较,1:4VCM组在培养60、72h时以及1:2VCM组在培养24h,RPE细胞的增生差异有统计学意义(P<0.05),而1:2VCM组在培养48、印、72h时RPE细胞的增生差异有非常统计学意义(P<0.01);1:2VCM组在培养各时间段混合细胞的增生差异有统计学意义(P<0.05),均表现为促增生效应.结论 一定体积分数的人VCM抑制HRCECs的增生,但明显促进人RPE细胞以及混合细胞的增生,提示玻璃体中RPE细胞浸润是加重外伤增生性玻璃体视网膜病变和眼内血管增生性疾病的高危因素.

Effects of human vitreous fluid on the proliferation of cultured human retinal capillary endothelial cells and pigment epithelial cells

Objective To investigate the effects of human vitreous fluid on proliferation of culturedhuman retinal capillary endothelial cells (HRCECs)and retinal pigment epithelial (RPE)cells.MethodsHRCECS and RPE cells were primary cultured and identified.Choose the P3 cells to be treated in differenthuman vitreous-conditioned medium (VCM)with or without serum, vitreous volume concentrations of VCMwere 1:8,1:4 and 1:2.Cells proliferation was assayed by tetrazolium (MTr)color imetry at the 24h,48h,60hand 72h respectively.Results In the presence of serum, 1:4,1:2 VCM had a strongly inhibitory effect onHRCECs proliferation compared with control group at all observed times retrospectively(P <0.05), but exerteda significantly stimulative effect on RPE cells and mixed cells (HRCECs mix RPE by 1:1)proliferationcompared with control group at all observed times retrospectively( P <0.01 ).In the absence of serum, 1:4 VCMat the 60,72h, and 1:2 VCM at the 24h had a stimulative effect on RPE cells growth compared with controlgroup(P <0.05), and 1:2 VCM at the 48,60,72h had a significantly stimulative effect on RPE cells(P<0.01);1:2 VCM had a significantly stimulative effect on mixed cells compared with control group at all observedtimes retrospectively (P <0.05 ).Conclusion Human vitreous fluid inhibits human HRCECS, but boosts RPEand mixed cells growth in vitro.This result suggests that RPE infiltration in vitreous may be a high risk factor intraumatic proliferative vitreo-retinopathy and intraocular neovascular disease.