Torricelli Paola, Fini Milena, Fanti Pier Alessandro, Dika Emi, Milani Massimo
Laboratory of Biocompatibility Innovative Technologies and Advanced Therapies (BITTA), Rizzoli Orthopaedic Institute, Bologna, Italy.
Dermatology Clinic of the University of Bologna, Bologna, Italy.
Photodermatol Photoimmunol Photomed. 2017 May;33(3):156-163. doi: 10.1111/phpp.12297. Epub 2017 Feb 23.
Polypodium leucotomos (PL) exerts potent antioxidant, photo-protective, and immune-modulatory activities. A reconstructed human epidermis (RHE) (Episkin) is a suitable model for the evaluation of acute UV-induced cell damage. No data regarding the photo-protective action of PL in this model are available.
We evaluated the effects of PL on the prevention of UVB-induced cell damage assessing sunburn cells, CPD formation, p53, Ki-67, p21 expression, and epidermal growth factor (EGF) production.
MATERIALS & METHODS: RHE was incubated in standard conditions. PL was topically applied at the concentration of 2 mg/cm , immediately before UVB exposition. UVB exposition (300 mJ/cm ) was performed using a dedicated UVB lamp. Irradiated samples without PL and non-irradiated samples were used as positive and negative controls. Expression of p53, p21, and Ki-67 was evaluated with immune-histochemical methods. CPD were measured using a monoclonal antibody.
PL significantly reduced sunburned cells (-80%) in comparison with positive control. PL significantly prevented the increase in EGF production at tested times. PL significantly reduced the p53 (-80%), p21 (-84%), and Ki-67 (-48%) positive cells. Finally, PL prevented the formation of CPD (0% vs. 20% positive cells).
In this model, PL has shown to prevent UVB cell damage, the upregulation of proliferating proteins, and fully blocking the formation of CPD.
白叶铁线蕨（PL）具有强大的抗氧化、光保护和免疫调节活性。重建的人表皮（RHE）（Episkin）是评估急性紫外线诱导细胞损伤的合适模型。目前尚无关于PL在此模型中光保护作用的数据。
我们通过评估晒伤细胞、环丁烷嘧啶二聚体（CPD）形成、p53、Ki-67、p21表达以及表皮生长因子（EGF）产生，来评估PL对预防紫外线B（UVB）诱导细胞损伤的作用。
RHE在标准条件下孵育。在UVB照射前，以2mg/cm²的浓度局部应用PL。使用专用UVB灯进行UVB照射（300mJ/cm²）。未使用PL照射的样本和未照射的样本分别用作阳性和阴性对照。采用免疫组织化学方法评估p53、p21和Ki-67的表达。使用单克隆抗体测量CPD。
与阳性对照相比，PL显著减少了晒伤细胞（-80%）。在测试时间内，PL显著抑制了EGF产生的增加。PL显著减少了p53（-80%）、p21（-84%）和Ki-67（-48%）阳性细胞。最后，PL阻止了CPD的形成（阳性细胞为0% vs. 20%）。
在此模型中，PL已显示出可预防UVB细胞损伤、增殖蛋白的上调，并完全阻止CPD的形成。
