【評(píng)價(jià)原理】
人類皮膚衰老的原因分為內(nèi)源性和外源性，外源性主要包括日曬、氧化和糖化。這里的糖化是指蛋白非酶糖基化反應(yīng)，即在非酶促條件下，蛋白質(zhì)、氨基酸、脂類或核酸等大分子物質(zhì)的游離氨基與還原糖的羰基經(jīng)過縮合、重排、裂解、氧化修飾等一系列反應(yīng)，最終形成晚期糖基化終產(chǎn)物（AGEs）。AGEs與皮膚衰老密切相關(guān)，過量的AGEs可以與皮膚彈性纖維、膠原蛋白發(fā)生糖化交聯(lián)反應(yīng)，并且AGEs呈棕色，最終使皮膚表現(xiàn)黃化、彈性降低。
常規(guī)的AGEs具有熒光特性，可利用葡萄糖與牛血清蛋白在體外發(fā)生糖化反應(yīng)生AGEs的原理進(jìn)行造模處理，加入抗糖化產(chǎn)品后通過熒光定量方法評(píng)價(jià)其抑制AGEs的水平來反映其抗糖化功效。
【實(shí)驗(yàn)方法】
將實(shí)驗(yàn)分為六個(gè)組，分別是正常對(duì)照組、模型對(duì)照組、陽性對(duì)照組、供試品低濃度組、供試品中濃度組和供試品高濃度組。選用牛血清蛋白作為基體，模型對(duì)照組加入葡萄糖溶液，陽性對(duì)照組加入葡萄糖和陽性對(duì)照藥，不同濃度供試品組加入葡萄糖和待檢測(cè)供試品，在60℃下孵育一定時(shí)間，用酶標(biāo)儀在370nm/440nm的熒光檢測(cè)條件下檢測(cè)AGEs的含量變化，從而反映待檢測(cè)供試品是否具有抗糖化功效。
【評(píng)價(jià)指標(biāo)】
AGEs熒光強(qiáng)度的變化
【結(jié)果展示】（展示圖片僅供參考，實(shí)際實(shí)驗(yàn)組別依據(jù)合同而定）
AGEs熒光強(qiáng)度的變化：提供柱狀圖
圖1. AGEs熒光強(qiáng)度的變化
【參考文獻(xiàn)】
建模方法及指標(biāo)應(yīng)用：
[1] Gkogkolou P, B?hm M. Advanced glycation end products: key players in skin aging?[J]. Dermato-endocrinology, 2012, 4(3): 259-270.
[2] Tseng JY, Ghazaryan AA, Lo W, Chen YF, Hovhannisyan V, Chen SJ, et al. Multiphoton spectral microscopy for imaging and quantification of tissue glycation. Biomed Opt Express 2010; 2:218-30;
[3] Zhang, G.; Huang, G.; Xiao, L.; Mitchell, A. E. Determination of advanced glycation endproducts by LC-MS/MS in raw and roasted almonds (Prunus dulcis). J. Agric. Food Chem. 2011, 59, 12037-12046.
[4] Lin J A, Wu C H, Yen G C. Perspective of advanced glycation end products on human health[J]. Journal of agricultural and food chemistry, 2018, 66(9): 2065-2070.
陽性藥應(yīng)用：
[5] Babizhayev MA, Nikolayev GM, Nikolayeva JG, Yegorov YE. Biologic activities of molecular chaperones and pharmacologic chaperone imidazole-containing
dipeptide-based compounds: natural skin care help and the ultimate challenge: implication for adaptive responses in the skin[J]. Am J Ther 2012; 19: e69-89.
[6] Babizhayev MA, Yegorov YE. Therapeutic uses of drug-carrier systems for imidazole-containing dipeptide compounds that act as pharmacological chaperones and have significant impact on the treatment of chronic diseases associated with increased oxidative stress and the formation of advanced glycation end products[J]. Crit Rev Ther Drug Carrier Syst 2010; 27:85-154.
浙公網(wǎng)安備 33010802003447號(hào)