Huang He, Jin Chong-Ying, Bi Xu-Kun, Zhao Yan-Bo, Xu Sheng-Jie, Wang Mei-Hui, Yu Lu, Sun Ya-Xun, Hu Dan
Department of Cardiology, Sir Run Run Shaw Hospital, College of Medicine, Zhejiang University, Hangzhou, China.
Department of Cardiology and Cardiovascular Research Institute, Renmin Hospital of Wuhan University, Wuhan, China.
Front Pharmacol. 2018 Nov 14;9:1305. doi: 10.3389/fphar.2018.01305. eCollection 2018.
Epigallocatechin gallate (EGCG) is the most abundant catechin in green tea and has proven benefits on endothelial cells in diabetes. However, it remains unclear whether EGCG could improve function of late endothelial progenitor cells (L-EPCs) in diabetes. Thirty-six rabbits were randomized into six groups. Thirty diabetic rabbits were induced by a single dose of alloxan (100 mg/kg injection intraperitoneally). All of them were given intragastrically EGCG (50 mg/kg/day) or saline for 7 days after carotid injury. In autotransfusion experiment, L-EPCs were cultured with pre-treated EGCG (40 μM for 72 h) and then were injected into the site of injured vascular. Proliferation and migration of EGCG pre-treated L-EPCs in high glucose condition were assessed by EDU incorporation assay and modified Boyden chamber assay, respectively. The mRNA and protein expression of Akt-eNOS pathway were detected by real-time PCR and western blot. Reendothelialization rate in injured carotid artery of diabetic rabbits was augmented in the EGCG group (50 mg/kg/d for 7 days) compared with the non-EGCG group (74.2 ± 4.6% vs. 25.6 ± 5.9%, < 0.001). EGCG pre-treated L-EPCs autologous transfusion also accelerated the diabetic rabbits' carotid reendothelialization compared with the diabetic sham-operated group (65.6 ± 8.5% vs. 32.9 ± 5.0%, = 0.011). studies showed, 40 μM EGCG treatment for 72 h recovered L-EPCs' proliferation and migration, as well as restored the phosphorylation level of Akt and eNOS blocked by high glucose condition. EGCG accelerated reendothelialization in diabetic rabbits after carotid injury in part by reactivating the Akt/eNOS pathway, which might contribute to recovering proliferation and migration of L-EPCs impaired by high glucose.
表没食子儿茶素没食子酸酯(EGCG)是绿茶中含量最丰富的儿茶素,已证实其对糖尿病患者的内皮细胞有益。然而,EGCG是否能改善糖尿病患者晚期内皮祖细胞(L-EPCs)的功能仍不清楚。36只兔子被随机分为6组。30只糖尿病兔子通过单次腹腔注射四氧嘧啶(100mg/kg)诱导产生。颈动脉损伤后,所有兔子均连续7天经胃给予EGCG(50mg/kg/天)或生理盐水。在自体输血实验中,将L-EPCs与预处理的EGCG(40μM,处理72小时)共同培养,然后注射到血管损伤部位。分别通过EDU掺入试验和改良的Boyden小室试验评估高糖条件下EGCG预处理的L-EPCs的增殖和迁移能力。通过实时PCR和蛋白质印迹法检测Akt-eNOS通路的mRNA和蛋白表达。与非EGCG组相比,EGCG组(50mg/kg/d,连续7天)糖尿病兔子损伤颈动脉的再内皮化率增加(74.2±4.6%对25.6±5.9%,P<0.001)。与糖尿病假手术组相比,EGCG预处理的L-EPCs自体输血也加速了糖尿病兔子颈动脉的再内皮化(65.6±8.5%对32.9±5.0%,P=0.011)。研究表明,40μM EGCG处理72小时可恢复L-EPCs的增殖和迁移能力,并恢复被高糖条件阻断的Akt和eNOS的磷酸化水平。EGCG通过部分激活Akt/eNOS通路加速糖尿病兔子颈动脉损伤后的再内皮化,这可能有助于恢复受高糖损害的L-EPCs的增殖和迁移能力。