摘要
目的研究藏红花素(Crocin)对过氧化氢(H2O2)诱导的人视网膜色素上皮(RPE)细胞凋亡的影响及可能机制。方法采用300μmol·L^-1的H2O2培养RPE细胞建立细胞氧化损伤模型,用10μmol·L^-1及100μmol·L^-1的藏红花素干预,观察藏红花素对在H2O2作用下RPE细胞活性、细胞内活性氧(ROS)水平、凋亡细胞比例和线粒体凋亡相关信号分子Bcl-2及Bax表达的干预。结果对照组RPE细胞存活率为98.33%,氧化损伤组RPE细胞存活率为43.12%,与对照组比较差异有统计学意义(F=48.08,P<0.05),不同浓度藏红花素干预后,RPE细胞存活率分别提高到61.76%和77.67%,与氧化损伤组比较差异均有统计学意义(F低=40.89,F高=90.33,均P<0.05);H2O2处理RPE细胞内ROS水平明显升高,10μmol·L^-1及100μmol·L^-1的藏红花素可不同程度抑制H2O2处理后RPE细胞内ROS水平;流式细胞仪的检测结果,与对照组(4.33%)比较,氧化损伤组RPE细胞凋亡率(63.12%)明显升高(F=89.02,P<0.05),随着藏红花素作用浓度的递增,H2O2诱导的RPE细胞凋亡率呈剂量依赖性递减(39.00%、18.00%),与氧化损伤组比较差异均有统计学意义(F低=192.40,F高=95.76,均P<0.05);此外,藏红花素可以改变细胞内Bcl-2及Bax的表达。结论藏红花素通过抑制RPE细胞内ROS生成、改变线粒体凋亡相关信号分子Bcl-2及Bax的表达,防止RPE细胞凋亡反应的发生。
OBJECTIVE To investigate the effect and possible mechanism of crocin on hydrogen peroxide(H2O2)-induced human retina pigment epithelium(RPE)apoptosis.METHODS The RPE cells oxidative damage model was induced by 300μmol·L^-1 H2O2,and intervened with 10μmol·L^-1 and100μmol·L^-1 of Crocin.Then RPE cell activity,intracellular reactive oxygen species(ROS)levels,apoptosis cell ratio and expressions of mitochondrial apoptosis signaling molecules as Bcl-2 and Bax were detected.RESULTS The survival rate of RPE in the control group was 98.33%,and 43.12%in the oxidative damage group,there was significant difference between two group(F=48.08,P<0.05).After the intervention of crocin in different concentration,the survival rate of RPE increased to 61.76%and 77.67%,respectively,and there were statistical differences between them and result of oxidative damage group(P<0.05);ROS generation in RPE increased obviously after H2O2 intervention,10μmol·L^-1 and 100μmol·L^-1 crocin both suppressed ROS generation levels within RPE.The results of flow cytometry showed that the apoptotic rate of RPE in oxidative injury group(63.12%)was significantly higher than result of 4.33%in control group(F=89.02,P<0.05).With the concentration of crocin increased,the apoptotic rate of RPE induced by hydrogen peroxide decreased in a dose-dependent manner(39.00%and 18.00%).Compared with result of oxidative damage group,the difference was significant(Flow=192.40,Fhigh=95.76,all P<0.05);In addition,crocin changed the expression of Bcl-2 and Bax within cells.CONCLUSIONS Crocin prevented the occurrence of RPE cells apoptosis response by inhibiting ROS generation in RPE cells,changing the expression of Bcl-2 and Bax which were mitochondrial apoptosis related signal molecules.
作者
李姚
王健
LI Yao;WANG Jian(First Affiliated Hospital of Harbin Medical University,Harbin 150001,China)
出处
《中国中医眼科杂志》
2019年第5期347-350,共4页
China Journal of Chinese Ophthalmology
