牡丹皮组分抑制“毒损因子AGEs”生成的作用及机制研究  

Role and mechanism of Cortex Moutan components in inhibiting production of toxic advanced glycation end products(AGEs)

作　　者：李杰[1] 陈婷婷 孙楠楠 杨冰 张梦桃 尹诗雨 张朱槟 张少婵 贾晓斌 封亮 LI Jie;CHEN Ting-ting;SUN Nan-nan;YANG Bing;ZHANG Meng-tao;YIN Shi-yu;ZHANG Zhu-bin;ZHANG Shao-chan;JIA Xiao-bin;FENG Liang(the First Affiliated Hospital of University of Science and Technology of China(Anhui Provincial Hospital),Hefei 230001,China;School of Traditional Chinese Pharmacy,China Pharmaceutical University,Nanjing 211198,China)

机构地区：[1]中国科学技术大学附属第一医院(安徽省立医院),安徽合肥230001 [2]中国药科大学中药学院,江苏南京211198

出　　处：《中国中药杂志》2022年第12期3215-3223,共9页China Journal of Chinese Materia Medica

基　　金：国家重点研发计划“中药现代化”专项(2018YFC1706900);国家自然科学基金项目(81973536);中国药科大学双一流创新团队项目(CPU2018GY11,CPU2018GF07)。

摘　　要：晚期糖基化终末产物(advanced glycation end products,AGEs)可导致糖尿病及其并发症等多种疾病。该研究构建体外非酶糖基化反应模型——牛血清白蛋白/丙酮醛(BSA/MGO reaction system)与牡丹皮提取物孵育,采用HPLC和UPLC-Q-TOF-MS/MS检测辨识牡丹皮提取物与丙酮醛共孵育液中抑制AGEs生成的活性成分,筛选出salvianan、paeoniside、苯甲酰芍药苷、牡丹苷J、没食子酰氧化芍药苷、氧化苯甲酰芍药苷、5-羟基-3S-羟甲基-6-甲基-2,3-二氢苯并呋喃、没食子酰芍药苷等差异性成分,推测其为牡丹皮提取物捕捉MGO的潜在活性成分;同时采用BSA-葡萄糖反应体系考察不同浓度的牡丹皮提取物(饮片质量浓度40、80、120、160、200 mg·mL^(-1))体外抑制AGEs生成的作用规律,进一步评价牡丹皮提取物及差异成分没食子酰芍药苷和苯甲酰芍药苷抑制高糖诱导的HUVEC内AGEs生成情况,并采用AO/EB双荧光染色法观察细胞凋亡。结果显示牡丹皮提取物及差异成分对AGEs的形成有一定的抑制作用,可降低细胞凋亡。该研究为牡丹皮通过抑制毒损因子AGEs治疗糖尿病血管并发症具有参考价值。Advanced glycation end products(AGEs)can lead to many diseases such as diabetes and its complications.In this study,an in vitro non-enzymatic glycosylation reaction model—bovine serum albumin/methylglyoxal(BSA/MGO)reaction system was constructed and incubated with Cortex Moutan extract.High performance liquid chromatography(HPLC)and ultra-high performance liquid chromatography with quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF-MS/MS)were used to detect and identify the active components that inhibited the formation of AGEs in the co-incubation solution of Cortex Moutan extract and MGO,and differential components such as salvianan,paeoniside,benzoylpaeoniflorin,mudanpioside J,galloyloxypaeoniflorin,benzoyloxy-paeoniflorin,5-hydroxy-3 s-hydroxymethyl-6-methyl-2,3-dihydro benzofuran,and galloylpaeoniflorin were screened out,which were inferred to be the potential active components of Cortex Moutan extract to capture MGO.In addition,BSA-glucose reaction system was performed to investigate the influence of different concentrations of Cortex Moutan extract(decoction concentrations:40,80,120,160,and 200 mg·mL^(-1))on inhibiting the production of AGEs in vitro.The inhibitory effects of Cortex Moutan extract and the differential components galloylpaeoniflorin and benzoyl paeoniflorin on the production of AGEs in human umbilical vein endothelial cells(HUVECs)induced by high glucose was further evaluated.Cell apoptosis was observed by acridine orange and ethidium bromide(AO/EB)double fluorescence staining.The results showed that Cortex Moutan Cortex extract and its differential components had certain inhibitory effects on the formation of AGEs,and could reduce cell apoptosis.This study provided reference for the treatment of diabetic vascular complications by Cortex Moutan inhibiting the toxic AGEs.

关 键 词：牡丹皮 组分 糖基化产物 血管内皮细胞 

分 类 号：R285[医药卫生—中药学]