作者
Linghong Guo,Yong Yang,Yiyao Pu,Shuangfa Mao,Yu Nie,Yin Liu,Xian Jiang
摘要
Dendrobium officinale Kimura & Migo is traditionally used to treat skin diseases, gastrointestinal diseases, and other diseases. Dendrobium officinale polysaccharides (DOP) are the main component of Dendrobium officinale that accounts for its bioactivity, which shows a variety of effects such as moisturizing, antioxidant and anti-fatigue. However, there is no comprehensive study on the effect of DOP on skin photoaging combined with in vitro and in vivo models, and its specific mechanism is still unclear.Our study aimed to explore the effect and underlying mechanism of DOP on skin photoaging, as well as to improve the stability and transdermal absorption of DOP.DOP was extracted, purified and structurally characterized. In vitro HaCaT cell photoaging model was used to examine the photoprotection effect of DOP. Cell viability was detected by CCK-8; Intracellular reactive oxygen species were determined by DCFH-DA; DNA damage, cell apoptosis and cell cycle arrest were examined by flow cytocytometry. For autophagy flux detection, the adenovirus loaded with mRFP-GFP-LC3 was introduced into cells. Further, to enhance the stability and absorption of DOP, we designed and prepared the W/O/W type DOP multilayer emulsions (ME) by a two-step emulsification method. The emulsion stability, drug loading and encapsulation rate, DOP stability and DOP transdermal rate were detected. In vivo photoaging animal model was applied to compare the difference of photoaging protection effect between DOP solution and DOP ME. Specifically, skin appearance, histological change, antioxidant system, proinflammatory indicators, matrix metalloproteinases and autophagy level of skin tissues were examined and compared.The results showed that DOP achieve photoaging protection by inhibiting oxidative stress, alleviating cell cycle arrest and apoptosis, and enhancing autophagy flux in photoaged HaCaT cells. The W/O/W type DOP multilayer emulsion (ME) with high encapsulation rate and strong stability was found to significantly improve the stability and transdermal absorption of DOP. In addition, our results showed that DOP (ME) remarkably improved skin condition of photoaged mice. Specifically, DOP (ME) enhanced the expression of antioxidant enzymes and autophagy and decreased the levels of pro-inflammatory factors and matrix metalloproteinases in the skin of photoaged mice as compared with DOP solution.In conclusion, DOP was effective in improving skin photoaging, and the DOP multilayer emulsion we designed enhanced the stability and skin absorption of DOP, boosting DOP's protective effect against photoaging.