陈丽娟,王加志,徐姣,齐永华,黄昕红

[摘要]目的：研究十精丸抗紫外线致ESF-1细胞光老化作用，并初步探讨其作用机制。方法：用40J/cm2的UVA照射人皮肤成纤维细胞建立光老化细胞模型，以不同浓度十精丸提取物处理光老化细胞，MTT法检测细胞活力，羟胺法检测细胞超氧化物歧化酶（SOD）活性，比色法检测谷胱甘肽过氧化物酶（GSH-Px）活性，TBA法检测丙二醛（MDA）含量。结果：UVA照射成纤维细胞后，细胞活力下降，SOD、GSH-Px活性降低，MDA含量升高（P<0.01），十精丸水提液中（100μg/ml）、高（200μg/ml）剂量组能显著提高成纤维细胞细胞活力、SOD、GSH-Px活性，降低MDA含量（P<0.05或P<0.01）。结论：十精丸水提液可抑制UVA引起的成纤维细胞活性下降，推测其机制为通过提高SOD活力、加速氧自由基的清除和减少氧自由基的产生，使细胞的脂质过氧化损伤程度降低有关。

[关键词]十精丸；UVA；ESF-1细胞；光老化

[中图分类号]Q813.1[文献标识码]A[文章编号]1008-6455（2011）04-0582-03

Protective effect of Shijingwan on photoaging induced by Ultraviolet A in ESF-1 cell

CHEN Li-juan,WANG Jia-zhi,XU Jiao,QI Yong-hua,HUANG Xin-hong

(Department of Cosmetology,Jiamusi College, Heilongjiang University of Traditional ChineseMedicine,Jiamusi 154007,Heilongjiang,China)

Abstract: ObjectiveTo study the protective effect of Shijingwan agains UV irradiation-induceddamage in ESF-1 cells and its mechanism.MethodsThe model of photoaging fibroblasts was established by irradiating of ultraviolet light(40J/cm2). The different concentration water-extract solution from Shijingwan were used for photoaging cell. The proliferation of human fibroblasts was detected by MTT, and superoxide dismutase(SOD) and glutathione peroxidase(GSH-Px) activity, malondialdehyde(MDA) content were assayed by hydroxylamine, colorimetric and TBA methods respectively.ResultsThe proliferation activity of fibroblasts descended with the UVA irradiation, the activities of SOD and GSH-Px decreased, when the content of MDA was ascensive(P<0.01). But under the intervention at middle concentration(100μg/ml) and high concentration(200μg/ml) of water-extract solution from Shijingwan,the above changes induced by UVA irradiation were inhibited(P<0.05 or P<0.01).ConclusionThe water-extract solution from Shijingwan might inhibit the proliferation decreasing which induced by UVA. And its mechanism is due to its abilities of increasing SOD，scavenging oxygen free radicals.

Keywords:Shijingwan; UVA; ESF-1cell; photoaging

皮肤长期暴露于紫外线（UV）下可导致结构及功能的改变，从而加速或导致皮肤老化的发生，这种因UV照射引起的老化被称为光老化。皮肤光老化不仅有损于人的美容，而且可能与皮肤癌的发生也有着病因学的联系[1]。随着当前生态环境的恶化，皮肤光老化现象愈来愈严重，严重影响着人们的身心健康。十精丸为补肾方药之一，出自明代《普济方·卷二百二十三·诸虚门·补虚驻颜色附论》，是中医美容保健、延年益寿之传统名方。但目前十精丸抗皮肤光老化的基础实验研究薄弱，严重限制了十精丸疗效的进一步发挥。笔者以体外培养人皮肤成纤维细胞ESF-1为研究对象，在体外成功建立ESF-1 UVA光老化模型，探讨十精丸水提液对UVA损伤ESF-1细胞的保护作用。

1材料和方法

1.1实验材料和仪器

1.1.1材料：皮肤成纤维细胞ESF-1（北京协和细胞中心），DMEM培养基（Gibco公司，批号：12800-058），胎牛血清（Billab公司，批号：20101030）、胰蛋白酶（Billab公司，批号：20100901）、双抗（Billab公司，批号：20101230），MTT（Sigma公司），超……