非瑟酮对人晶状体上皮细胞增殖和凋亡的影响

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（ｈｕｍａｎｌｅｎｓｅｐｉｔｈｅｌｉａｌｃｅｌｌ，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１２ｈ及２４ｈ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（１２ｈ、２４ｈ后分别为０．１１７６±０．０１５０和０．１１７２±０．００６１），凋亡率明显增加（２４ｈ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１２ｈ及２４ｈ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２４ｈ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０．３９９４±０．０２５７）（Ｐ＜０．０５）。培养２４ｈ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。

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

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 H202 0xidative damage stress . the collected cells were divided into normal control group , H2 02 group , Fis group , Fis + H202 group. According to the concentration of Fis (5 yg . mL-’,IO yg . mL-l and 20 yg . mL ’1 ) .the Fis group and Fis + H202 groups were divided int0 3 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 and 24 hours , compared with the control group , many cells of H2 02 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 sigruficant differences ( all P < 0. 01) . At the 12 hours and 24 hours.the morphology and the proliferation of Fis groups didnt change evidently (P > 0. 05 ). Compared with the H202 group , the Fis + H202 group appeared less morphology change and the cell proliferation was improved in a time and dose-de-pendent manner ( the peak was 0. 3994 + 0. 0257 ) ( P < 0. 05 ) . Compared with the H2 02 group , the apoptotic rate in Fis + H202 group with different dose of Fis were ( 9. 99 +1. 53 ) % , ( 5. 80 + 1. 55 ) % , ( 3. 58 + 0. 73 ) % , respectively , there was statistical difference (P < 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.