组合体外与临床测试评价植物抗氧化剂抗老化功效

Skin antiaging efficacy evaluation for plant antioxidants integrating in vitro and clinical test

目的基于非动物测试理念,研究植物抗氧化剂复配后的皮肤抗老化功效。方法体外培养HaCaT角质细胞,紫外线照射诱发氧化损伤模型,分别加入白藜芦醇、原花青素和二者复配物,用MTT比色法、荧光素标记法和ELISA法检测细胞活性、活性氧簇自由基(ROS)、超氧化物岐化酶(SOD)、细胞周期及细胞凋亡的变化。对含两种植物原料的眼霜配方进行鸡胚尿囊膜血管试验(CAMVA)和牛角膜混浊渗透试验(BCOP)的组合测试评价其眼刺激性。选择平均年龄40岁的女性志愿者25名,眼部连续使用该眼霜60 d,于使用产品的第0、30和60 d分别检测皮肤的水份、水份流失、弹性和肤色等皮肤老化相关指标。结果白藜芦醇-原花青素复配的植物抗氧化剂可明显缓解紫外线造成的HaCaT细胞氧化损伤,SOD水平明显升高(P<0.001),ROS荧光强度、细胞凋亡率及S期细胞比例均明显降低(P<0.001),细胞修复作用强于单独作用。经过替代方法组合评价其安全性后进行临床志愿者测试,证明皮肤保湿、弹性和皮肤颜色等指标得到改善。结论白藜芦醇和原花青素两种植物抗氧化剂复配使用可有效减少皮肤氧化应激和明显改善皮肤性能。

Objective To investigate the skin antiaging efficacy of plant antioxidant blend based on the concept of non-animal testing.MethodsHaCaT keratinocyte cell line were induced oxidation damage by ultraviolet irradiation and treated with resveratrol, procyanidins and two substances blend respectively. The quantitative measurement including cell viability, ROS, SOD, cell cycle and cell apoptosis were detected by MTT assay, fluorescein labeling and ELISA method. The ocular irritation of eye cream formula containing plant material were assessed by combination of CAMVA and BCOP testing. Clinical efficacy was evaluated in terms of chronoaging and photoaging features including skin hydration, elasticity and color by non-invasive methods on 25female volunteers after 60days of treatment. ResultsResveratrol-procyanidins blend antioxidant could significantly relieve HaCaT cell oxidative lesion induced by UV. The SOD level in test group were significantly higher than radiation group （P〈0.001）. All ROS fluorescence intensity, apoptosis cell and the percentage of S-phase cell in cell cycle were obviously decreased in test group （P〈0.001）. The antioxidant ability of blend are better than resveratrol and procyanidins. The clinical test of volunteers after alternative combination evaluation of its safety, which showed that skin moisturization, TEWL, elasticity and color had significantly improved. ConclusionSpecific plant antioxidant blend by resveratrol and procyanidins could effectively reduce skin oxidative stress and improve function.