RGDRGD肽对人晶状体上皮细胞黏附与增生作用的影响

背景RGD肽是一类含有精氨酸-甘氨酸-天冬氨酸的小分子多肽，其在抑制肿瘤细胞黏附、转移和肿瘤新生血管的生成中起重要作用。研究证实RGD肽能够抑制晶状体上皮细胞（LECs）的黏附和增生，RGDRGD肽串联起来作用增强。目的研究RGDRGD肽对体外培养的人LECs黏附与增生的影响，并与RGD肽的作用进行比较。方法将在体外分离培养的LECs中分别加入250、500、1000mg／L的RGDRGD肽和RGD肽作为实验组，仅加入细胞培养液作为对照组。将LECs以2×10。／ml密度接种到含有纤维连接蛋白（FN）和I型胶原蛋白预孵化的96孔培养板中，培养1h后用MTT比色法检测RGDRGD肽与RGD肽对细胞黏附率的影响。将LECs接种于培养板，分别加入250、500、1000、2000mg／L的RGDRGD肽和RGD肽培养24、48、72h，检测RGDRGD肽和RGD肽对LECs增生的影响。结果RGDRGD肽对人LECs黏附率的抑制呈明显的剂量依赖性，随着其质量浓度的增加，对细胞黏附的抑制作用越强，500mg／L的RGDRGD肽比RGD肽抑制人LECs黏附的作用更强（P〈0．01）。RGDRGD肽对人LECs增生的抑制呈明显的时间剂量依赖性，1000mg／L的RGDRGD肽作用48h比RGD肽对人LECs增生的抑制作用更强（P〈0．01）。结论RGDRGD肽抑制LECs黏附与增生的作用强于RGD肽，为进一步临床应用提供了理论依据。

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

Background RGD is a small molecular multiple peptide containing Arg-Gly-Asp with an important role in inhibiting the adhesion,migration and neovascularization of tumor.Our previous study determined that RGD can suppress the adhesion and proliferation of lens epithelial cells(LECs),and RGDRGD may be of a stronger effect. Objective Present study was to investigate and compare the effect of RGDRGD peptide on the proliferation and adhesion of immortalized human LECs(HLEB-3)in vitro. Methods Human LECs harvested by trpsin-EDTA were suspended in DMEM medium with serial dilutions of RGD peptide and RGDRGD peptide(from 1000 mg/L to 250 mg/L)at 37℃ for 15 minutes as experimental group,and the HLECs cultured by common culture medium were used as the control group.The cells were then seeded into the 96-well plates with precoated fibmnectin (FN)and I collagen at the density of 2×104/ml.MTT stainingcolorimetry was used to measure the adhesion rates of lens epithelial cells cultured in different concentrations RGDRGD and RGD peptides after 1 hour.Cells were seeded into the 96-well plates for 24 hours at 37℃ in 5% CO2.Medium was then replaced with DMEM overnight.Subsequently,the cells were treated with serial dilutions of RGD and RGDRGD(from 2000 mg/L to 250 ms/L)dissolved in DMEM medium plus 20% fetal bovine serum.The inhibition of RGDRGD and RGD on the adhesion and proliferation of Human LECs was analyzed by MTT aher 24,48 and 72 hours. Results The inhibition rate of RGD peptide on the adhesion of LECs was gradually enhanced with the increase of concentration with the significant difference among the different concentrations groups(F=1089.56,P＜0.01),and the statistically significant elevation in inhibitory rate was found in RGDRGD peptide compared with RGD peptide(P＜0.01).The inhibition rate of RGDRGD peptide on the proliferation of LECs wag gradually increased with the increase of concentration with the significant difference among the different concentrations groups with a strongest effect in 1000 ms/L group(F=127.31,P＜0.01),and the much stronger inhibition Wag Been in RGDRGD peptide(F=1589.85,P＜0.01).The suppression rate of RGDRGD on LECs proliferation Wag much stronger with the prolong of time(F=1606.43,P＜0.01). Conclusion RGDRGD peptide and RGD peptide have inhibitory effect on adhesion and proliferation of human LECs in a dose-and time-dependent manner.Effect of RGDRGD peptide is much stronger than RGD peptide.These results imply that RGDRGD peptide and RGD peptide have the important role for prevention of PCO.