非瑟酮对人晶状体上皮细胞增殖和凋亡的影响

Effects of fisetin on proliferation and apoptosis of human lens epithelial cells

目的研究非瑟酮(fisetin,Fis)在生理状态下及氧化应激状态下对人晶状体上皮细胞(human lens epithelial cell,HLEC)增殖和凋亡的影响。方法体外培养HLEC,通过H2O2氧化损伤建立氧化应激模型,设置空白对照组、H2O2组、Fis组和Fis+H2O2组,其中Fis组和Fis+H2O2组按Fis浓度(5μg·mL-1、10μg·mL-1和20μg·mL-1)分为3个亚组。分别于培养12 h及24 h后,倒置相差显微镜下观察各组细胞的形态学改变,运用MTT法检测细胞增殖的变化,运用流式细胞技术检测细胞凋亡率的变化。结果与空白对照组比较,H2O2组较多细胞出现典型的形态学改变,细胞增殖能力明显降低(12 h、24 h后分别为0.117 6±0.015 0和0.117 2±0.006 1),凋亡率明显增加(24 h后为12.35%±1.23%),差异均有统计学意义(均为P<0.01)。不同浓度Fis组间的细胞在培养12 h及24 h后细胞形态均无明显改变,细胞增殖也无明显变化(P>0.05)。培养12及24 h后,与H2O2组比较,Fis+H2O2组发生形态改变的细胞减少,细胞增殖能力明显改善,且随时间、Fis浓度增加其作用更明显(最高为0.399 4±0.025 7)(P<0.05)。培养24 h后,与H2O2组凋亡率比较,不同浓度Fis+H2O2组的细胞凋亡率逐渐降低,依次为(9.99±1.53)%、(5.80±1.55)%、(3.58±0.73)%,差异有统计学意义(P<0.05)。结论一定浓度的Fis在一定时段对生理状态下的HLEC增殖无明显影响。在氧化应激状态下,Fis呈时间和浓度依赖性地改善HLEC增殖能力,呈浓度依赖性地降低HLEC凋亡率。

Objective To investigate the effects of fisetin（ Fis） on the proliferation and apoptosis of cultured human lens epithelial cells（ HLEC） under the physiologic condition or oxidative injury. Methods After HLEC was cultured in vitro,the oxidative damage model was established through the H2O2 oxidative damage stress,the collected cells were divided into normal control group,H2O2 group,Fis group,Fis ＋ H2O2 group. According to the concentration of Fis（ 5 μg·mL- 1, 10 μg·mL- 1 and 20 μg·mL- 1）,the Fis group and Fis ＋ H2O2 groups were divided into3 subgroups. After cultured for 12 hours and 24 hours,growth condition and morphologic feature in each group were observed under invert microscope. The cell viability was assayed by MTT. The cell apoptotic rate was determined by flow cytometry. Results At the 12 hours and24 hours, compared with the control group,many cells of H2O2 group appeared typical morphology of apoptosis,the cell proliferation was obviously decreased（ 0. 117 6 ± 0. 015 0 and 0. 117 2± 0. 006 1）,the apoptotic rate was significantly increased（ 12. 35% ± 1. 23% at 24 hours）,there were significant differences（ all P ＆lt; 0. 01）. At the 12 hours and 24 hours,the morphology and the proliferation of Fis groups didn ’t change evidently（ P ＆gt; 0. 05）. Compared with the H2O2 group,the Fis ＋ H2O2 group appeared less morphology change and the cell proliferation was improved in a time and dose-dependent manner（ the peak was 0. 3994 ± 0. 0257）（ P ＆lt; 0. 05）.Compared with the H2O2 group,the apoptotic rate in Fis ＋ H2O2 group with different dose of Fis were（ 9. 99 ± 1. 53） %,（ 5. 80 ± 1. 55） %,（ 3. 58 ± 0. 73） %, respectively,t here was statistical difference（ P ＆lt; 0. 05）. Conclusion Under the physiologic condition, fis does not affect the proliferation of HLEC in a certain time and concentration. On the condition of oxidative stress,Fis improves the proliferation of HLEC in a time and dose-dependent manner, and decrease the apoptotic rate of HLEC in a dose- dependent manner.