Perez S, Sergent O, Morel P, Chevanne M, Dubos M P, Cillard P, Cillard J
Laboratoire de Biologie Cellulaire, INSERM U 49, Faculté de Pharmacie, France.
C R Seances Soc Biol Fil. 1995;189(3):453-65.
Lipid peroxidation has been implicated in skin damage by ultraviolet radiation. The aim of the study was to determine the kinetic of lipid peroxidation induced by ultraviolet beta (UVB) in adult keratinocytes and fibroblasts in culture. The keratinocytes were obtained from a single primary culture and the fibroblasts were in the same subculture (4 to 10 transfers). For UVB irradiation, the cells were maintained in a small volume of Hanks balanced salt solution and were irradiated (0.75, 1.5, 3 and 4.5 Jcm-2). Then cells were cultured for 3 to 48 hours. Lipid peroxidation was estimated by free MDA determination in both extracellular medium and cells using a size exclusion chromatography coupled to an HPLC procedure. In addition, LDH release in culture media was evaluated as in indice of cytotoxicity. An increase of total free MDA was observed 3 hours after cell irradiation which was dose-dependent from 0.75 to 3 Jcm-2 for keratinocytes and fibroblasts. MDA was detected both in cells and in culture media. As soon as 3 hours after irradiation 90% in total MDA was present in the culture media. Kinetic of lipid peroxidation: for 0.75 Jcm-2, an elevation of MDA was observed 12 hours after irradiation in both cultures. A further increase in MDA was noted 24 hours after fibroblasts irradiation but not in irradiated keratinocytes. LDH release in culture media increased with post irradiation time until 48 hours. The cytotoxic effect of UVB irradiation on keratinocytes and fibroblasts cultures was shown by an enhancement of lipid peroxidation which was detectable during 48 hours after irradiation. An increase of LDH release was observed simultaneously.
脂质过氧化作用与紫外线辐射导致的皮肤损伤有关。本研究的目的是确定培养的成年角质形成细胞和成纤维细胞中，紫外线β（UVB）诱导的脂质过氧化作用的动力学。角质形成细胞取自单一原代培养物，成纤维细胞处于同一传代培养（4至10次传代）。对于UVB照射，将细胞维持在少量的汉克斯平衡盐溶液中并进行照射（0.75、1.5、3和4.5 J/cm²）。然后将细胞培养3至48小时。使用与HPLC程序联用的尺寸排阻色谱法，通过测定细胞外培养基和细胞中的游离丙二醛（MDA）来评估脂质过氧化作用。此外，评估培养基中乳酸脱氢酶（LDH）的释放作为细胞毒性指标。细胞照射后3小时观察到总游离MDA增加，对于角质形成细胞和成纤维细胞，从0.75至3 J/cm²呈剂量依赖性。在细胞和培养基中均检测到MDA。照射后3小时，培养基中就存在90%的总MDA。脂质过氧化作用的动力学：对于0.75 J/cm²，两种培养物在照射后12小时均观察到MDA升高。成纤维细胞照射后24小时MDA进一步增加，但照射的角质形成细胞中未出现。培养基中LDH的释放在照射后48小时内随时间增加。UVB照射对角质形成细胞和成纤维细胞培养物的细胞毒性作用表现为脂质过氧化作用增强，在照射后48小时内均可检测到。同时观察到LDH释放增加。
