三叶鬼针草花提取物对甲基乙二醛诱导的蛋白质糖基化和 DNA 氧化损伤的保护作用。
Protective role of Clitoria ternatea L. flower extract on methylglyoxal-induced protein glycation and oxidative damage to DNA.
机构信息
Program in Veterinary Biosciences, Faculty of Veterinary Sciences, Chulalongkorn University, Bangkok, 10330, Thailand.
Phytochemical and Functional Food Research Unit for Clinical Nutrition, Department of Nutrition and Dietetics, Faculty of Allied Health Sciences, Chulalongkorn University, Bangkok, 10330, Thailand.
出版信息
BMC Complement Med Ther. 2021 Mar 1;21(1):80. doi: 10.1186/s12906-021-03255-9.
BACKGROUND
Methylglyoxal (MG) is a highly reactive dicarbonyl precursor for the formation of advanced glycation end products (AGEs) associated with age-related diseases, including diabetes and its complications. Clitoria ternatea L. flower has been reported to possess antioxidant and antiglycating properties. Evidence indicates that the extract of Clitoria ternatea L. flower inhibits fructose-induced protein glycation and oxidative damage to bovine serum albumin (BSA). However, there is no evidence to support the inhibitory effect of CTE against MG-mediated protein glycation and oxidative damage to protein and DNA. Therefore, the aim of the present study was to investigate whether C. ternatea flower extract (CTE) prevents MG-induced protein glycation and oxidative DNA damage.
METHODS
The formation of fluorescent AGEs in BSA was evaluated using spectrofluorometer. The protein carbonyl and thiol group content were used for detecting protein oxidation. DNA strand breakage in a glycation model comprising of MG, lysine and Cu or a free radical generator 2,2'-azobis(2-methylpropionamidine) dihydrochloride (AAPH) systems was investigated using gel electrophoresis. Generation of superoxide anions and hydroxyl radicals in the MG/lysine system was assessed by the cytochrome c reduction assay and thiobarbituric acid reactive substances assay, respectively. High performance liquid chromatography (HPLC) was used to measure the MG-trapping ability.
RESULTS
In the BSA/MG system, CTE (0.25-1 mg/mL) significantly inhibited the formation of fluorescent AGEs and protein oxidation by reducing protein carbonyl content as well as preventing the protein thiol depletion. The concentration of CTE at 0.125-1 mg/mL prevented oxidative DNA cleavage in MG/lysine and AAPH systems associated with the inhibition of superoxide anion and hydroxyl radical formation. It also directly trapped MG in a concentration-dependent manner, ranging from 15 to 43%.
CONCLUSIONS
The study findings suggest that the direct carbonyl trapping ability and the free radical scavenging activity of CTE are the underlying mechanisms responsible for the prevention of protein glycation and oxidative DNA damage.
背景
甲基乙二醛 (MG) 是一种高度反应性的二羰基前体，可形成与年龄相关的疾病相关的晚期糖基化终产物 (AGEs)，包括糖尿病及其并发症。已报道三叶鬼针草花具有抗氧化和抗糖化作用。有证据表明，三叶鬼针草花提取物可抑制果糖诱导的蛋白质糖化和牛血清白蛋白 (BSA) 的氧化损伤。然而，没有证据支持 CTE 对 MG 介导的蛋白质糖化和蛋白质及 DNA 氧化损伤的抑制作用。因此，本研究旨在探讨三叶鬼针草花提取物 (CTE) 是否可预防 MG 诱导的蛋白质糖化和氧化 DNA 损伤。
方法
使用荧光分光光度计评估 BSA 中荧光 AGEs 的形成。使用蛋白羰基和巯基含量检测蛋白氧化。使用电泳法研究 MG、赖氨酸和 Cu 或自由基生成剂 2,2'-偶氮双（2-甲基丙脒）二盐酸盐 (AAPH) 系统中的糖基化模型中的 DNA 链断裂。通过细胞色素 c 还原测定法和硫代巴比妥酸反应物质测定法分别评估 MG/赖氨酸系统中超氧阴离子和羟基自由基的产生。使用高效液相色谱法 (HPLC) 测量 MG 捕获能力。
结果
在 BSA/MG 系统中，CTE（0.25-1mg/mL）通过降低蛋白羰基含量以及防止蛋白巯基耗竭，显著抑制荧光 AGEs 的形成和蛋白氧化。浓度为 0.125-1mg/mL 的 CTE 可防止 MG/赖氨酸和 AAPH 系统中与超氧阴离子和羟基自由基形成相关的氧化 DNA 断裂。它还以浓度依赖的方式直接捕获 MG，范围为 15 至 43%。
结论
研究结果表明，CTE 的直接羰基捕获能力和自由基清除活性是预防蛋白质糖化和氧化 DNA 损伤的潜在机制。
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