Lim Wonbong, Ko Mikyung, Lee Sungga, Kim Inae, Jung Mina, Kim Okjoon, Cho Seonghoun, Yang Kyuho, Choi Namki, Kim Sunmi, Choi Hongran
Department of Oral Pathology, Second Stage of Brain Korea 21 for School of Dentistry, Dental Science Research Institute, Chonnam National University, Bug-Gu, Gwangju, Korea.
Photomed Laser Surg. 2008 Jun;26(3):215-20. doi: 10.1089/pho.2007.2142.
The purpose of this study was to examine the protection afforded by 635-nm irradiation against ultraviolet (UV)-C-induced apoptosis in primary human gingival fibroblasts (hGFs).
UV irradiation is known to cause photoaging and cellular apoptosis of skin cells and is considered to be one of the leading causes of skin carcinogenesis.
To induce apoptosis, UV-C (100 mJ/cm2) was used to irradiate hGFs. To protect them from apoptosis, pretreatment with 635-nm irradiation was performed for 1 h immediately after cell plating 36 or 48 h before UV-C irradiation. The light source used for irradiation was a continuous-wave 635-nm LED laser emitting at 1 mW/cm2. Experimental samples were selected 24 h after UV-C irradiation. To measure the numbers of apoptotic cells, MTT assay and flow cytometric analyses were performed. For histomorphologic findings, Diff-Quick staining was carried out. Also, the activities and mRNA expression of caspase-3, caspase-8, and caspase-9 were measured.
In the present study, the number of apoptotic cells declined in the cells that were pretreated with 635-nm light irradiation in a time-dependent manner. In addition, the activities and mRNA expression of caspase-3, caspase-8, and caspase-9 were significantly recovered by pretreatment with 635-nm irradiation.
These results suggest that 635-nm visible light irradiation may be used as a protective tool to prevent UV-C-induced apoptosis.
本研究旨在探讨635纳米照射对紫外线(UV)-C诱导的原代人牙龈成纤维细胞(hGFs)凋亡的保护作用。
已知紫外线照射会导致皮肤细胞光老化和细胞凋亡,被认为是皮肤癌发生的主要原因之一。
为诱导凋亡,用UV-C(100 mJ/cm2)照射hGFs。为保护细胞免受凋亡,在UV-C照射前36或48小时细胞接种后立即用635纳米照射预处理1小时。用于照射的光源是发射功率为1 mW/cm2的连续波635纳米LED激光。在UV-C照射后24小时选取实验样本。为测量凋亡细胞数量,进行MTT法和流式细胞术分析。对于组织形态学观察,进行Diff-Quick染色。此外,还测量了半胱天冬酶-3、半胱天冬酶-8和半胱天冬酶-9的活性及mRNA表达。
在本研究中,经635纳米光照射预处理的细胞中凋亡细胞数量呈时间依赖性下降。此外,经635纳米照射预处理后,半胱天冬酶-3、半胱天冬酶-8和半胱天冬酶-9的活性及mRNA表达显著恢复。
这些结果表明,635纳米可见光照射可作为预防UV-C诱导凋亡的一种保护手段。