葛根素微脂體製備與其對皮膚光老化的抑制作用

老化現象一直以來都是一個重要的議題，皮膚老化日益受到關注，近來許多皮膚抗老化產品推陳出新。本研究的目的在於探討紫外線引起之皮膚光老化，主要機制為發炎、氧化壓力、DNA損傷等。本研究利用天然萃取物葛根素抗氧化特性做為抗皮膚老化藥物。葛根素在沒有藥物載體的情況下生物利用率低，因此利用微脂體做為葛根素之藥物載體應用於皮膚遞送系統上，提高生物利用率。本研究利用有機溶劑從植物野葛根萃取葛根素後以固相萃取法純化，可以得到異黃酮類純度98.5 %、HPLC測試葛根素純度75.5 ± 2.36 %、MS測試葛根素純度33.085 ± 0.95 %。利用ANOVA分析得到微脂體最佳比例為E6C2T2P2，載藥量達16.23 %，粒徑在140 nm左右。在蔗糖與EGG PC重量比30：1之蔗糖溶液環境下置備微脂體後冷凍乾燥，粒徑改變率為1.20 %，說明此置備方式成功製作微脂體凍乾試劑。葛根素、葛根素微脂體與微脂體凍乾試劑作用於HaCaT細胞上有效抑制了UVB誘導的凋亡與發炎反應，且葛根素微脂體與凍乾試劑以較低劑量500 ng/mL達到與高劑量葛根素200 μg/mL相同的抑制效果。並且確定了紫外線誘導凋亡路經由活性氧化物質(ROS)介導JNK訊息傳遞路徑誘導細胞凋亡，藉由ROS介導MAPKs、PI3K/Akt、PKC家族與NF-κB等訊息傳遞路徑誘導細胞呈現發炎狀態。

關鍵字

皮膚老化；凋亡；發炎；葛根素；微脂體

並列摘要

Aging has always been a big issue for human being, and there is growing concern about the problem of cutaneous aging. Therefore, cosmetic products preventing cutaneous aging has increased significantly. This study focuses on photoaging, which is caused by UV radiation. It has been well known that photoaging includes several pathways, such as, oxidative stress, inflammation and apoptosis. In this study, puerarin, a natural extract from Pueraria lobata, is used to prevent photoaging. In view of the poor bioavailability of puerarin without a drug carrier, a liposomal dosage has been chosen to increase the biovailability of puerarin. Organic solvent is used to extract the puerarin, and solid phase extraction is used to purify the puerarin. The purity of isoflavone is found to be 98.5 % and the purity of puerarin is found to be 75.5 ± 2.36 % by HPLC, 33.085 ± 0.95 % by MS. ANOVA analysis is used to optimize the ratio between the components of liposome, which is found to be E6C2T2P2. Sucrose shows that minimal particle size changes after the freeze-dry study, while the weight ratio of sucrose to EGG PC is 30 : 1. Delivering puerarin, liposomal puerarin and proliposome of puerarin into HaCaT cell effectively inhibits inflammation and apoptosis induced by UVB. In addition, using low concentrations of liposome dosage can reach the same expectation as using high concentrations of puerarin. The role of UVB-induced apoptosis and inflammation was found that UVB up-regulation of MAPKs, PI3K/Akt, PKC, NF-κB and CPLA2 pathway via reactive oxygen species (ROS).

並列關鍵字

Skin aging, Apoptosis ； Inflammation ； Puerarin ； Liposome

參考文獻

72. 陳竺音, 葛根素微脂體於大鼠腎系膜細胞之體外抗氧化評估. 2013.

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葛根素微脂體製備與其對皮膚光老化的抑制作用

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