Department of Ophthalmology, Longhua Hospital, Shanghai University of Traditional Chinese Medicine, 725 Southern Wanping Road, Shanghai 200032, China; School of Chinese Medicine, the University of Hong Kong, 10 Sassoon Road, Pokfulam, Hong Kong, China.
School of Chinese Medicine, the University of Hong Kong, 10 Sassoon Road, Pokfulam, Hong Kong, China.
J Ethnopharmacol. 2015 Jul 1;169:295-304. doi: 10.1016/j.jep.2015.04.031. Epub 2015 Apr 28.
He-Ying-Qing-Re Formula (HF) is a formula modified from "Si-Miao-Yong-An Decoction", a traditional Chinese medical classic emerged in the Qing dynasty and has been reported for treatment of vascular diseases. HF, containing 8 herbs, has been used in local hospital for decades as a complementary method for diabetic retinopathy (DR) with retinal vascular dysfunction. Clinical reports revealed HF could ameliorate vision defects, microaneurysms, hemorrhages and macular edema. The aim of this study is to investigate the anti-DR action of HF and its underlying mechanism experimentally.
Chromatographic fingerprinting of HF and rodent model of DR were established; hypoglycemic effect of HF was measured by fasting, random blood glucose and glucose tolerance test; vascular degeneration was measured by retinal digestion; blood-retina-barrier (BRB) permeability was assessed with Evans Blue leakage assay. Advanced glycation end products (AGEs) were measured in vitro and in vivo level; Migration of retinal vascular endothelial cells were determined by wound healing and transwell chamber assays; permeability of endothelial monolayer was monitored with dextran transport. AGEs-related proteins and signaling were measured with immunoblotting and immunohistochemistry.
Chlorogenic acid, ferulic acid and arctin were identified as major components in HF; HF suppresses retinal vasculature degeneration and BRB permeability damage without significant inhibition on hyperglycemia; HF reduces in vitro and in vivo formation of AGEs and AGEs-induced migration as well as permeability of retinal vascular endothelial cells. Expression of tight junction proteins Zo-1 and Claudin-1 was increased while activation of AGEs receptor and downstream signaling Akt were suppressed upon HF treatment.
HF exhibits protective effect against diabetic retinopathy, which may be associated with inhibition on AGEs and recovery on endothelial dysfunction via modulation of tight junction and AGEs downstream signaling.
草药配方 HF 对糖尿病视网膜病变的作用及其机制研究
药用相关性：和营清血方（HF）是由清朝经典名方“四物汤”加减而来，用于治疗血管疾病。HF 包含 8 种草药，在当地医院被用于治疗糖尿病视网膜病变（DR）及其视网膜血管功能障碍已数十年，作为一种辅助治疗方法。临床报告表明 HF 可以改善视力缺陷、微动脉瘤、出血和黄斑水肿。本研究旨在通过实验研究探讨 HF 对 DR 的防治作用及其作用机制。
建立 HF 的色谱指纹图谱和 DR 动物模型；通过空腹、随机血糖和糖耐量试验测定 HF 的降血糖作用；通过视网膜消化法测定血管退行性变；通过 Evans Blue 渗漏试验评估血视网膜屏障（BRB）通透性。在体内和体外测定晚期糖基化终产物（AGEs）；通过划痕愈合和 Transwell 室测定实验测定视网膜血管内皮细胞迁移；通过葡聚糖转运监测内皮单层通透性。通过免疫印迹和免疫组化测定 AGEs 相关蛋白和信号转导。
鉴定出 HF 的主要成分有绿原酸、阿魏酸和苍术苷；HF 可抑制视网膜血管退化和 BRB 通透性损伤，而对高血糖无明显抑制作用；HF 可减少体内外 AGEs 的形成以及 AGEs 诱导的迁移和视网膜血管内皮细胞的通透性。HF 处理后，紧密连接蛋白 Zo-1 和 Claudin-1 的表达增加，而 AGEs 受体及其下游信号 Akt 的激活受到抑制。
HF 对糖尿病视网膜病变具有保护作用，其机制可能与抑制 AGEs 的形成和恢复内皮功能障碍有关，通过调节紧密连接和 AGEs 下游信号。
