CAJ | 학술논문

目的探讨姜黄素对兔视网膜色素上皮(RPE)细胞增生的抑制作用及其作用机制.方法选取第4代RPE细胞进行实验,分为姜黄素组和空白对照组,姜黄素组设10、15、20 μg/ml 3个质量浓度.噻唑蓝比色(MTT)法检测姜黄素10、15、20μg/ml分别在24、48、72、96 h对体外培养的RPE细胞增生的抑制率.线性回归分析计算各时间段的半数抑制率(IC50)剂量.流式细胞仪(FMC)检测姜黄素15 μg/ml作用72 h后对RPE细胞周期的影响,并且检测姜黄素15μg/ml作用24、48,72 h后RPE细胞增生细胞核抗原(PCNA)的表达量和凋亡率.透射电子显微镜进行RPE细胞形态学检测.结果姜黄素24、48、72、96 h的IC50分别是29.31,17.50、13.24、10.99 μg/ml.姜黄素使RPE细胞阻滞在G0/G1期.姜黄素15 μg/ml作用24、48、72 h后,RPE细胞的PCNA表达量分别为(565.04±23.60),(473.61±36.88),(396.15±32.45),凋亡率分别为(12.83±0.13)%,(32.27±4.51)%,(56.81±8.67)%,各浓度组与对照组相比,差异均有统计学意义(P<0.05).透射电子显微镜显示RPE细胞出现凋亡特征.结论姜黄素可以影响RPE细胞的PCNA合成,诱导其凋亡,从而有效抑制RPE细胞增生.姜黄素有望成为一种有效的防治PVR的天然药物.
목적 탐토강황소대토시망막색소상피(RPE)세포증생적억제작용급기작용궤제.방법 선취제4대RPE세포진행실험,분위강황소조화공백대조조,강황소조설10、15、20 μg/ml 3개질량농도.새서람비색(MTT)법검측강황소10、15、20μg/ml분별재24、48、72、96 h대체외배양적RPE세포증생적억제솔.선성회귀분석계산각시간단적반수억제솔(IC50)제량.류식세포의(FMC)검측강황소15 μg/ml작용72 h후대RPE세포주기적영향,병차검측강황소15μg/ml작용24、48,72 h후RPE세포증생세포핵항원(PCNA)적표체량화조망솔.투사전자현미경진행RPE세포형태학검측.결과 강황소24、48、72、96 h적IC50분별시29.31,17.50、13.24、10.99 μg/ml.강황소사RPE세포조체재G0/G1기.강황소15 μg/ml작용24、48、72 h후,RPE세포적PCNA표체량분별위(565.04±23.60),(473.61±36.88),(396.15±32.45),조망솔분별위(12.83±0.13)%,(32.27±4.51)%,(56.81±8.67)%,각농도조여대조조상비,차이균유통계학의의(P<0.05).투사전자현미경현시RPE세포출현조망특정.결론 강황소가이영향RPE세포적PCNA합성,유도기조망,종이유효억제RPE세포증생.강황소유망성위일충유효적방치PVR적천연약물.
Objective To observe the inhibition effect of curcumin on the proliferation of rabbit retinal pigment epithelial (RPE) cells and investigate its mechanism.Methods The 4th generation of RPE cells were selected and divided into curcumin group and blank control group.The concentration of curcumin included 10,15,and 20 μg/ml.The MTT assay was used to evaluate the inhibition effect on the proliferation of RPE cells at the 24th,48th,72nd and 96th hour after cultured with curcumin (10,15,and 20 μg/ml).The IC50 value of curcumin at different time points were calculated by Linear Regression.Flow cytometry was used to detect the effect on the cell cycle at the 72nd hour after cultured with eurcumin (15 μg/ml) ;the expression and apoptosis of proliferating cell nuclear antigen (PCNA) were also determined at the 24th,48th,and 72nd hour after cultured with curcumin (15 μg/ml) respectively.The configuration of RPE cells were observed by transmission electron microscope.Results The IC50 value of curcumin at the 24th,48th,72nd and 96th hour was 29.31,17.50,13.24,and 10.99 μg/ml respectively.Cell cycel analysis indicated that cureumin blocked cells in G0/G1 phase.At the 24th,48th,and 72nd hour after cultured with curcumin (15 μg/ml),the expression of PCNA of RPE cells were 565.04±23.60,473.61± 36.88,and 396.15±32.45;the apoptosisrate were (12.83±0.13)%,(32.27±4.51)%,(56.81± 8.67)%,respectively.The differeces of curcumin groups compared with the control group were significant (P<0.05).Apoptosis of RPE cells was observed under transmission electron microscope.Conclusions Curcumin can inhibite the proliferation of RPE cells by inhibit the synthesization of PCNA and inducing the apoptosis of RPE cells.Curcumin may become a potential drug to prevent and treat PVR.