人表皮角质形成细胞急性光损伤模型的构建

Construction of acute light damage model with human epidermal keratinocytes

目的探讨长波紫外线(UVA)和中波紫外线(UVB)对人永生化角质形成细胞(HaCaT细胞)造成急性损伤所需要的最小剂量,为构建人表皮细胞急性光损伤模型提供科学依据。方法用不同剂量的UVA或UVB照射HaCaT细胞。观察细胞形态学改变,检测细胞增殖活性(吸光度)和凋亡率。结果 2.5～10 J/cm2UVA照射对HaCaT细胞无损伤,20～40 J/cm2可致细胞出现形态学改变,与空白对照组相比增殖活性降低,凋亡率增加(P<0.01);11.4～34.2 mJ/cm2UVB照射对HaCaT细胞无损伤,随照射剂量增加,细胞出现不同程度损伤,与空白对照组相比增殖活性降低,凋亡率增加(P<0.01)。结论 UVA和UVB照射均可构造急性光损伤模型。HaCaT细胞对UVB敏感性更高,引起急性光损伤所需的最小剂量更低。

Objective To explore a minimum dose of ultraviolet A （UVA） and ultraviolet B （UVB） that could cause immortalized keratinocytes （HaCaT cells） acute injury, and provide a scientific basis for building human epidermal cell acute light damage model. Methods HaCaT cells were treated with different dose of UVA / UVB. The morphological changes, as well as proliferation activity and apoptosis rate were ob- served. Result No injury changes were onservd with radiating dose of low radiation dose as 2. 5 - 10 J/cm2 UVA. Yet 20 - 40 J/era2 caused cellular morphological changes, with decreased proliferative activity and in- creased apoptosis rate （P 〈 0. 01 ）. No injury ehanges were observed with radiating dose of 11.4 - 34. 2 mJ/ cm2 UVB. However, cells manifested with varied morphological changes, activity and apoptosis rate with te in- vrease of UVB dosage （P 〈 0. 01 ）. Conclusion Both UVA and UVB radiation can be used as construction to make acute light damage model. HaCaT cells presented with more sensitivity with UVB, showing lower mini- mum required dose to cause acute light damage. As a dose-dependent model, type and degree of radiations were alternative in different experimental settings.