桃红四物汤对糖尿病视网膜病变引起的细胞损伤及HIF-1α表达的影响  

Taohong Siwu Decoction alleviates cell damage caused by diabetic retinopathy by down-regulating the expression of HIF-1α

作　　者：王磊[1] 俞莹[2] 陆萍 程燕妮 李沭岩 WANG Lei;YU Ying;LU Ping;CHENG Yanni;LI Shuyan(The Seventh People’s Hospital,Shanghai University of Traditional Chinese Medicine,Shanghai 200137,China;Shuguang Hospital,Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China;Shanghai Guangde Clinic of Traditional Chinese Medicine,Shanghai 200125,China)

机构地区：[1]上海中医药大学附属第七人民医院,上海200137 [2]上海中医药大学附属曙光医院,上海201203 [3]上海广德中医门诊部,上海200125

出　　处：《现代中西医结合杂志》2023年第21期2921-2929,共9页Modern Journal of Integrated Traditional Chinese and Western Medicine

基　　金：国家自然科学基金项目(82104636);上海市卫生健康委员会项目(20204Y0005);上海市浦东新区卫生系统名中医继承人培养计划(PWRzj2020-24);上海中医药大学预算内科研项目(2021LK060)。

摘　　要：目的探究桃红四物汤对高糖诱导的人视网膜微血管内皮细胞(hRMECs)的保护作用及潜在的分子机制。方法①取hRMECs,设置5组:空白对照组加5 mmol/L葡萄糖培养,高糖模型组加30 mmol/L葡萄糖培养,桃红四物汤低、中、高剂量组分别加30 mmol/L葡萄糖和相应浓度桃红四物汤含药血清培养,通过Transwell和划痕实验检测细胞的侵袭和迁移能力,血管生成实验检测细胞的成管能力,ELISA试剂盒检测细胞中血管内皮生长因子(VEGF)水平,Western blot法检测细胞中缺氧诱导因子-1α(HIF-1α)蛋白表达情况。②另外设置4组:空白对照组加5 mmol/L葡萄糖培养,高糖模型组加30 mmol/L葡萄糖培养,桃红四物汤组加30 mmol/L葡萄糖和高剂量桃红四物汤含药血清培养,桃红四物汤+HIF-1α过表达组采用HIF-1α过表达的hRMECs细胞加30 mmol/L葡萄糖和高剂量桃红四物汤含药血清培养,考察HIF-1α过表达对桃红四物汤的功能回复作用。结果①与空白对照组比较,高糖模型组细胞侵袭和迁移能力均明显增强(P均<0.05),成管数量明显增多(P<0.05),VEGF水平和HIF-1α蛋白相对表达量均明显增加(P均<0.05)。与高糖模型组比较,桃红四物汤高剂量组细胞的侵袭和迁移能力均明显降低(P均<0.05),成管数量明显减少(P均<0.05),VEGF水平和HIF-1α蛋白相对表达量均明显降低(P均<0.05)。②与高糖模型组比较,桃红四物汤组细胞的侵袭和迁移能力均明显降低(P均<0.05),成管数量明显减少(P均<0.05),VEGF水平和HIF-1α蛋白相对表达量均明显降低(P均<0.05);与桃红四物汤组比较,桃红四物汤+HIF-1α过表达组细胞侵袭和迁移能力均明显增强(P均<0.05),VEGF水平和HIF-1α蛋白相对表达量均明显升高(P均<0.05)。结论桃红四物汤通过下调HIF-1α的表达减轻糖尿病视网膜病变引起的细胞损伤。Objective It is to explore the protective effect of Taohong Siwu Decoction(THSWD)on high glucose-induced human retinal microvascular endothelial cells(hRMECs)and its potential molecular mechanism.Methods①hRMECs were taken and some of which were divided into 5 groups:the blank control group was cultured with 5 mmol/L glucose,the high-glucose model group was cultured with 30 mmol/L glucose,and the low,medium,and high-dose groups of THSWD were cultured with 30 mmol/L glucose and THSWD drug-containing serum of the corresponding concentration,respectively,the invasive and migratory abilities of the cells were detected by Transwell and scratch assays,the tubularization ability of the cells was detected by angiogenesis assay,the level of vascular endothelial growth factor(VEGF)in the cells was detected by ELISA kit,and the protein expression of hypoxia-inducible factor-1α(HIF-1α)in the cells was detected by Western blot assay.②Some of the cells were additionally divided into 4 groups:the blank control group was cultured with 5 mmol/L glucose,the high-glucose model group was cultured with 30 mmol/L glucose,the THSWD group was cultured with 30 mmol/L glucose and high-dose THSWD containing serum,and the THSWD+HIF-1αoverexpression group was cultured with 30 mmol/L glucose and high-dose THSWD containing serum,in which the hRMECs cells were HIF-1αoverexpressed,in order to investigate the functional recovery effects of HIF-1αoverexpression on THSWD.Results①Compared with the blank control group,the invasion and migration abilities of the cells in the high-glucose model group were significantly increased(all P<0.05),the number of tube forming was significantly increased(P<0.05),and the relative protein expressions of VEGF and HIF-1αwere significantly increased(all P<0.05).Compared with the high-glucose model group,the invasion and migration abilities of hRMECs cells in the high-dose THSWD group were significantly decreased(all P<0.05),the number of tube forming was significantly decreased(P<0.05),the relative protein

关 键 词：糖尿病视网膜病变 桃红四物汤 缺氧诱导因子 血管内皮生长因子 人视网膜微血管内皮细胞 

分 类 号：R-332[医药卫生]