Amar Saroj Kumar, Goyal Shruti, Dubey Divya, Srivastav Ajeet K, Chopra Deepti, Singh Jyoti, Shankar Jai, Chaturvedi Rajnish K, Ray Ratan Singh
Photobiology Division, CSIR-Indian Institute of Toxicology Research, P.O. Box 80, M.G. Marg, Lucknow 226001, India; Academy of Scientific and Innovative Research, CSIR-IITR Campus, Lucknow, India.
Photobiology Division, CSIR-Indian Institute of Toxicology Research, P.O. Box 80, M.G. Marg, Lucknow 226001, India.
Toxicol Lett. 2015 Dec 15;239(3):182-93. doi: 10.1016/j.toxlet.2015.09.024. Epub 2015 Oct 9.
Solar UV radiation is main factor of photocarcinogenesis, photoageing, and phototoxicity; thus, protection from UV radiation is major concern. Sunscreens containing UV filters are suggested as sun safe practices, but safety of UV filters remains in controversies. Benzophenone-1 (BP1) is commonly used in sunscreens as UV blocker. We assessed the photogenotoxicity and apoptotic parameters in human keratinocytes (HaCaT cells) by western blot, immunocytochemistry, flowcytometry, comet assay and TEM imaging. Our results exposed that BP1 photosensitized and generated intracellular ROS (2.02 folds) under sunlight/UVR. Decrease in cell viability was recorded as 80.06%, 60.98% and 56.24% under sunlight, UVA and UVB, respectively. Genotoxic potential of BP1 was confirmed through photomicronuclei and CPDs formation. BP1 enhanced lipid peroxidation and leakage of LDH enzyme (61.7%). Apoptotic cells were detected by AnnexinV/PI staining and sub G1 population of cell cycle. BP1 induced up regulation of apoptotic proteins Bax/Bcl2 ratio, Apaf-1, cytochrome c, Smac/DIABLO and cleaved caspase 3 was noticed. Down regulation of pro caspase 3 was inhibited by Z-VAD-fmk (inhibitor of caspase). Thus, study established the involvement of BP1 in photogenotoxicity and apoptosis via release of cytochrome c and Smac/DIABLO. These findings suggest sunscreen user to avoid BP1 in cosmetics preparation for its topical application.
太阳紫外线辐射是光致癌、光老化和光毒性的主要因素；因此，防止紫外线辐射是主要关注点。含有紫外线过滤剂的防晒霜被建议作为防晒安全措施，但紫外线过滤剂的安全性仍存在争议。二苯甲酮-1（BP1）常用于防晒霜中作为紫外线阻滞剂。我们通过蛋白质免疫印迹法、免疫细胞化学、流式细胞术、彗星试验和透射电镜成像评估了人角质形成细胞（HaCaT细胞）中的光遗传毒性和凋亡参数。我们的结果表明，BP1在阳光/紫外线照射下会发生光致敏并产生细胞内活性氧（增加2.02倍）。在阳光、UVA和UVB照射下，细胞活力分别下降了80.06%、60.98%和56.24%。通过光致微核和环丁烷嘧啶二聚体（CPDs）的形成证实了BP1的遗传毒性潜力。BP1增强了脂质过氧化和乳酸脱氢酶（LDH）的泄漏（61.7%）。通过膜联蛋白V/碘化丙啶染色和细胞周期亚G1期群体检测到凋亡细胞。发现BP1诱导凋亡蛋白Bax/Bcl2比值、凋亡蛋白酶激活因子-1（Apaf-1）、细胞色素c、第二线粒体衍生激活因子/直接 IAP 结合蛋白低分子量蛋白（Smac/DIABLO）和裂解的半胱天冬酶-3上调。Z-VAD-fmk（半胱天冬酶抑制剂）抑制了原半胱天冬酶-3的下调。因此，该研究确定了BP1通过细胞色素c和Smac/DIABLO的释放参与光遗传毒性和凋亡。这些发现表明，防晒霜使用者应避免在化妆品制剂中局部使用BP1。
