Sakamoto Asuka, Nakamura Masatsugu
Ophthalmic Research & Development Center, Santen Pharmaceutical Co., Ltd., Ikoma, Nara, Japan.
Yakugaku Zasshi. 2012;132(8):933-7. doi: 10.1248/yakushi.132.933.
This study evaluated the effects of flavin adenine dinucleotide (FAD) on ultraviolet B (UV-B)-induced damage in cultured human corneal epithelial (HCE-T) cells. The cultured HCE-T cells were treated with 0.003125-0.05% FAD before exposure to 80 mJ/cm2 UV-B. Cell viability was measured 24 h after UV-B irradiation using the MTS assay. Reactive oxygen species (ROS) were detected 30 min after UV-B irradiation using 5-(and-6)-chloromethyl-2',7'-dichlorodihydrofluorescein diacetate acetyl ester. Apoptosis was evaluated 4 h after UV-B irradiation in the caspase-3/7 activity assay. UV-B irradiation reduced cell viability and stimulated ROS production and caspase-3/7 activity in HCE-T cells. Pretreatment of UV-B irradiated HCE-T cells with FAD significantly attenuated cell viability reduction and inhibited the stimulation of both ROS production and caspase-3/7 activity due to UV-B exposure compared with those with vehicle (0% FAD). These results clarified that FAD inhibits ROS-mediated apoptosis by UV-B irradiation in HCE-T cells and suggest that FAD may be effective as a radical scavenger in UV-B-induced corneal damage.
本研究评估了黄素腺嘌呤二核苷酸（FAD）对培养的人角膜上皮（HCE-T）细胞中紫外线B（UV-B）诱导损伤的影响。在暴露于80 mJ/cm2 UV-B之前，用0.003125 - 0.05%的FAD处理培养的HCE-T细胞。使用MTS法在UV-B照射后24小时测量细胞活力。使用5-(和-6)-氯甲基-2',7'-二氯二氢荧光素二乙酸乙酰酯在UV-B照射后30分钟检测活性氧（ROS）。在UV-B照射后4小时通过caspase-3/7活性测定评估细胞凋亡。UV-B照射降低了HCE-T细胞的活力，并刺激了ROS的产生和caspase-3/7活性。与用载体（0% FAD）处理的细胞相比，用FAD预处理UV-B照射的HCE-T细胞显著减轻了细胞活力的降低，并抑制了由于UV-B暴露引起的ROS产生和caspase-3/7活性的刺激。这些结果表明，FAD抑制UV-B照射在HCE-T细胞中诱导的ROS介导的细胞凋亡，并表明FAD在UV-B诱导的角膜损伤中可能作为自由基清除剂有效。
