姜黄素抑制人晶状体上皮细胞增殖与胶原蛋白合成的研究

Inhibitive effects of curcumin on proliferation and collagen synthesis of human lens epithelial cell

目的:探讨中药单体姜黄素(Cur)对人晶状体上皮细胞系(HLE-B3)增殖及细胞内Ⅰ型和Ⅲ型胶原蛋白合成的影响。方法:20mg/L的Cur作用于HLE-B3 24h后MTT法检测吸光度值(OD值),苏木精-伊红染色观察细胞形态,流式细胞术检测细胞周期变化,ELISA法检测Ⅰ型和Ⅲ型胶原蛋白表达。结果:rhbFGF组OD值较正常组升高,Cur组OD值较rhbFGF组降低,抑制率达53.70%;Cur组细胞数目明显减少,胞浆减少,轮廓不清,交织的网状结构减少,甚至有的细胞变圆,细胞核凝集,见核固缩现象,胞浆嗜酸性染色;G1期的细胞Cur组较rhbFGF组明显增加,S期的细胞Cur组较rhbFGF组明显减少,G2期的细胞Cur组较rhbFGF组无明显变化;Ⅰ型胶原蛋白浓度为Cur组较rhbFGF组明显下降,Ⅲ型胶原蛋白浓度Cur组较rhbFGF组也明显下降。结论:Cur抑制rhbFGF诱导的HLE-B3增殖的同时也能抑制HLE-B3内Ⅰ、Ⅲ型胶原蛋白合成,干扰HLE-B3纤维化,可望成为防治后囊膜混浊的理想药物。

Objective： To investigate the effects of Curcumin（Cur） on proliferation and collagen synthesis of human lens epithelial cells B3（HLE-B3）.Methods： Recombinant human basic fibroblast growth factor（rhbFGF） was utilized to induce proliferation of HLE-B3.Proliferative HLE-B3 was incubated with 20 mg/L Cur in CO2 incubator for 24 hours.Then the inhibitory effect of Cur on proliferation of HLE-B3 was evaluated by methyl thiazolyl tetrazolium（MTT）.The effect of Cur on HLE-B3 morphology was observed under the optical microscope.The effect of Cur on HLE-B3 cell cycle was analyzed by flow cytometer（FCM）.Then the contents of collagen Ⅰ and collagen Ⅲ of HLE-B3 were assayed by enzyme-linked immunosorbent assay（ELISA）.Results： MTT test showed that the optical density（OD） value of rhbFGF group was remarkably higher than that of control group,and that of Cur group was obviously declined in comparison to rhbFGF group.The inhibitory rate of Cur was 53.70 %;In proliferation group,the number of HLE-B3 was increased with the normal cell structure and abundant cytoplasm under the optical microscope.However,in Cur group,the number of HLE-B3 was evidently decreased with less cytoplasm,undistinguished cell structure,condensed and aggregated nucleuses;The result of flow cytometer showed that the percentage of HLE-B3 in G1 phase in Cur group was evidently increased in comparison with rhbFGF group.HLE-B3 in S phase in Cur group was lower than rhbFGF group HLE-B3 in G2 phase in Cur group was no difference than rhbFGF group;The Collagen I contents of HLE-B3 was（24.58±1.68）μg/L in Cur,which decreased obviously to compare with rhbFGF group.The Collagen III contents of HLE-B3 were（0.18±0.05）μg/L in Cur groups,which decreased obviously to compare with rhbFGF group.Conclusion： Cur could effectively inhibit HLE-B3 proliferation induced by rhbFGF.Cur could inhibite the synthesis of collagen I and collagen III in HLE-B3,as well.The inhibitive effects of Cur on the HLE-B3 proliferation and the synthesis of collagen I and collagen III make it probably become reliable and effective drugs to prevent and treat posterior capsular opacification.