利用体外测评方法评价供试品抗糖化功效

【评价原理】

人类皮肤衰老的原因分为内源性和外源性，外源性主要包括日晒、氧化和糖化。这里的糖化是指蛋白非酶糖基化反应，即在非酶促条件下，蛋白质、氨基酸、脂类或核酸等大分子物质的游离氨基与还原糖的羰基经过缩合、重排、裂解、氧化修饰等一系列反应，最终形成晚期糖基化终产物（AGEs）。AGEs与皮肤衰老密切相关，过量的AGEs可以与皮肤弹性纤维、胶原蛋白发生糖化交联反应，并且AGEs呈棕色，最终使皮肤表现黄化、弹性降低。

常规的AGEs具有荧光特性，可利用葡萄糖与牛血清蛋白在体外发生糖化反应生AGEs的原理进行造模处理，加入抗糖化产品后通过荧光定量方法评价其抑制AGEs的水平来反映其抗糖化功效。

【实验方法】

将实验分为六个组，分别是正常对照组、模型对照组、阳性对照组、供试品低浓度组、供试品中浓度组和供试品高浓度组。选用牛血清蛋白作为基体，模型对照组加入葡萄糖溶液，阳性对照组加入葡萄糖和阳性对照药，不同浓度供试品组加入葡萄糖和待检测供试品，在60℃下孵育一定时间，用酶标仪在370nm/440nm的荧光检测条件下检测AGEs的含量变化，从而反映待检测供试品是否具有抗糖化功效。

【评价指标】

AGEs荧光强度的变化

【结果展示】（展示图片仅供参考，实际实验组别依据合同而定）

AGEs荧光强度的变化：提供柱状图

图1. AGEs荧光强度的变化

【参考文献】

建模方法及指标应用：

[1] Gkogkolou P, Böhm 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.

阳性药应用：

[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.