RGD肽对体外培养的人晶状体上皮细胞粘附与增殖的影响

Influence of RGD peptide on the proliferation and adhesion of human lens epithelial cells in vitro

目的:研究RGD肽对体外培养的人晶状体上皮细胞粘附与增殖的影响,为RGD肽防治晶状体后囊混浊提供理论依据。方法:将在体外分离培养的人晶状体上皮细胞中分别加入系列浓度的RGD肽(50,125,250,500,1000mg/L)作为实验组,不含RGD肽的培养基作为对照组。以2×107个/L密度接种到预孵化含有纤维连接蛋白和I型胶原蛋白的96孔培养板中,于1h后应用MTT法检测RGD肽对细胞粘附的影响。细胞接种于培养板,加入系列浓度RGD肽(125,250,500,1000,2000mg/L)后的24,48,72h检测对细胞增殖的影响。结果:RGD肽对人晶状体上皮细胞粘附的抑制呈明显的剂量依赖性,随着其浓度增大,细胞粘附数就越低,500mg/L时,抑制作用最强(P<0.05);RGD肽对人晶状体上皮细胞增殖的抑制呈明显的时间剂量依赖性,在48h,1000mg/LRGD条件下,对细胞的抑制作用最强。结论:RGD肽可抑制晶状体上皮细胞的粘附与增殖,具有潜在的防治后囊混浊的作用。

AIM： To study the effect of RGD peptide on the proliferation and adhesion of immortalized human lens epithelial cells （HLEB-3） in culture in order to provide theoretical basis for treatment of posterior capsular opacification （PCO）. METHODS. Human lens epithelial cells （HLECs） harvested by trypsin-EDTA were suspended in DMEM medium with serial dilutions of RGD peptide（from 50mg/L to 1 000mg/L ） at 37℃ for 15 minutes as experimental group, and the common culture medium as the control. Then cells were seeded into the 96-well plates with preincubated fibronectin （ FN ） and Ⅰ collagen at the density of 2 × 10^7/L. MTT colorimetry was used to measure the adhesion rates of lens epithelial cells cultured in different concentrations of RGD peptides after 1 hour. Cells were seeded into the 96-well plates for 24 hours at 37℃ in 50mL/L CO2. Medium was then replaced with DMEM overnight. Subsequently , cells were treated with serial dilutions of RGD （from 125mg/L to 2 000mg/L ） dissolved in DMEM medium plus 200mL/L fetal bovine serum . The inhibition of proliferation was analyzed by MTT after 24, 48 and 72 hours respectively. RESULTS： The inhibition of lens epithelial cells adhesion by RGD was dose-dependent ; The number of attached cells decreased with the increasing of RGD concentrations and peaking at 500mg/L （ P 〈 0.05）. The inhibition of HLECs＇ proliferative ability by RGD was time and dose dependent and peaking at 1 000mg/L （cultured for 48 hours）. CONCLUSION. RGD peptide can inhibit the adhesion and proliferati on of HLECs in vitro , and therefore it is potentially useful for preventing PCO.