光甘草定包合物对中波紫外线诱导小鼠皮肤光损伤的保护作用

Protection of glabridin inclusion on middlewave ultraviolet radiation induced skin photo damage in mice

目的 以环糊精(cyclodextrin,CD)为载体制备光甘草定(glabridin,GL)包合物,探讨光甘草定环糊精包合物(GL/CD)对中波紫外线(middlewave ultraviolet radiation, UVB)诱导的小鼠皮肤光损伤的保护作用。方法 制备GL/CD,进行包封率和透射电子显微镜(transmission electron microscope, TEM)表征分析,通过UVB照射小鼠皮肤光损伤模型,在小鼠背部皮肤涂抹不同浓度的GL及GL/CD,通过测试经皮水分散失(trans-epidermal water loss, TEWL)、皮肤组织生化指标、免疫组化染色研究其对光损伤后皮肤屏障保护的效果。结果 GL/CD的包封率为84.21%±1.36%,平均粒径为(533.1±42.8)nm,Zeta电位为(-30.24±1.54)m V,表明包合物在水体系中分散为纳米颗粒且稳定性良好。TEM结果显示GL/CD分散在水体系后主要呈现为球形胶束状态。小鼠皮肤黑色素染色结果显示GL/CD对黑色素生成的抑制效果优于GL。TEWL测试结果显示GL/CD显著降低小鼠经皮失水率,且效果比GL好。苏木精-伊红染色结果显示GL经CD包合之后能显著增强其抗炎作用,降低UVB照射引起的表皮增厚和炎症浸润。酶联免疫吸附实验结果显示GL经CD包合之后能显著提升抗氧化酶包括超氧化物歧化酶和过氧化氢酶的生物合成,降低活性氧及促炎细胞因子包括白细胞介素1、白细胞介素6和肿瘤坏死因子α在体内的表达水平。结论 GL经过CD的包合,有效提升了其在UVB诱导的皮肤损伤方面的保护作用。

Objective To synthesize glabridin(GL)inclusion complexes utilizing cyclodextrin(CD)as a delivery system,and elucidate the protective effects of the glabridin-cyclodextrin inclusion complexes(GL/CD)against middlewave ultraviolet radiation(UVB)-induced photo damage in murine skin.Methods The GL/CD complexes were formulated and subsequently characterized using encapsulation efficiency assays and transmission electron microscopy(TEM).A UVB irradiation-induced photo damage model in murine skin was constructed.Varying concentrations of GL and GL/CD were applied topically to the dorsal skin of mice.Protective effects on the skin barrier post UVB-induced damage were evaluated through measurements of trans-epidermal water loss(TEWL),biochemical indices of skin tissue,and immunohistochemical staining.Results The encapsulation efficiency of GL/CD was recorded as 84.21%±1.36%,with an average particle size of(533.1±42.8)nm and a Zeta potential of(-30.24±1.54)mV,indicating that the complexes dispersed in aqueous systems as nanoparticles and exhibited favorable stability.TEM images revealed that GL/CD primarily displayed as spherically micellar formations in water systems.Analysis of melanin staining in the mouse skin indicated that GL/CD more effectively inhibited melanogenesis compared to GL alone.TEWL assessments illustrated that GL/CD significantly decreased the rate of transepidermal water loss in mice,outperforming GL alone.Hematoxylin&Eosin staining indicated that the anti-inflammatory action of GL was notably enhanced post CD-encapsulation,significantly reducing the epidermal thickening and inflammatory infiltration caused by UVB irradiation.Enzyme linked immunosorbent assay results showed that post CD-encapsulation,GL notably elevated the biosynthesis of antioxidative enzymes,including superoxide dismutase and catalase,reduced reactive oxygen species,and lowered the expression levels of pro-inflammatory cytokines such as interleukin-1,interleukin-6,and tumor necrosis factor-alpha.Conclusion The encapsulation of GL with CD improves its protective effects against UVB-induced skin damage.