Muzaffer Umar, Paul V I, Prasad Nagarajan Rajendra, Karthikeyan Ramasamy
Department of Zoology, Faculty of Science, Annamalai University, Annamalai Nagar 608002, Tamilnadu, India.
Department of Biochemistry and Biotechnology, Faculty of Science, Annamalai University, Annamalai Nagar 608002, Tamilnadu, India.
Biochem Biophys Rep. 2018 Jan 28;13:109-115. doi: 10.1016/j.bbrep.2018.01.004. eCollection 2018 Mar.
The present study was aimed to investigate the photoprotective effect of the male flower of L. (MEJR) against ultraviolet-B induced apoptosis in human skin cells. Human skin epidermal keratinocytes were pretreated with the MEJR (80 µg/ml, has been selected after MTT assay), prior to 30 min UVB-irradiation at a dose of 20 mJ/cm. Mitochondrial membrane potential was evaluated using Rhodamine-123 staining; the % apoptosis by Hoechst staining and acridine orange staining; DNA damage was measured by comet assay. The levels of p53, Bax, Bcl-xL, Bcl-2, Cytochrome c, Caspase-9 and Caspase-3 expression in HaCaT cells were analyzed by western blotting and RT-PCR. Pretreatment with MEJR 80 µg/ml prior to UVB-irradiation significantly prevents apoptotic characteristics, DNA damage and loss of mitochondrial membrane potential. Thus, MEJR protects UVB-mediated human skin cells, by modulating the expression of apoptotic markers and UVB-induced DNA damage in HaCaT cells.
本研究旨在探讨光叶楮雄花(MEJR)对紫外线B诱导的人皮肤细胞凋亡的光保护作用。在以20 mJ/cm的剂量进行30分钟紫外线B照射之前,用人皮肤表皮角质形成细胞进行MEJR(80 μg/ml,经MTT试验后选定)预处理。使用罗丹明-123染色评估线粒体膜电位;通过Hoechst染色和吖啶橙染色评估凋亡百分比;通过彗星试验测量DNA损伤。通过蛋白质免疫印迹法和逆转录-聚合酶链反应分析HaCaT细胞中p53、Bax、Bcl-xL、Bcl-2、细胞色素c、半胱天冬酶-9和半胱天冬酶-3的表达水平。在紫外线B照射之前用80 μg/ml的MEJR预处理可显著预防凋亡特征、DNA损伤和线粒体膜电位丧失。因此,MEJR通过调节凋亡标志物的表达和紫外线B诱导的HaCaT细胞DNA损伤来保护紫外线B介导的人皮肤细胞。