基于HIF-1α通路探讨芍药苷改善糖尿病视网膜病变引起的组织和细胞损伤的机制  

The mechanism of paeoniflorin improving tissue and cell damage caused by diabetes retinopathy through the HIF-1α pathway

作　　者：刘霞 易勉 李灵[1] 岳江[1] 赵静 罗杏梅 黄洁 LIU Xia;YI Mian;LI Ling;YUE Jiang;ZHAO Jing;LUO Xingmei;HUANG Jie(Department of Ophthalmology,Changsha Fourth Hospital,Changsha 410006,Hunan Province,China)

机构地区：[1]长沙市第四医院眼科,湖南省长沙市410006

出　　处：《眼科新进展》2025年第3期196-201,共6页Recent Advances in Ophthalmology

基　　金：湖南省卫生健康委员会科研项目(编号:20230273)。

摘　　要：目的 探讨缺氧诱导因子-1α(HIF-1α)通路对芍药苷改善糖尿病视网膜病变引起的组织和细胞损伤的可能机制。方法 将30只大鼠随机分为3组[对照组(正常大鼠)、DR组(糖尿病模型大鼠)及芍药苷组(糖尿病模型大鼠且给予80 mg·kg^(-1)芍药苷灌胃)],每组各10只。培养大鼠视网膜微血管内皮细胞(rRMECs)并分为3组[对照组(5 mmol·L^(-1)葡萄糖培养)、高糖组(30 mmol·L^(-1)葡萄糖培养)及芍药苷组(30 mmol·L^(-1)葡萄糖+20 mol·L^(-1)芍药苷培养)],均混合培养24 h。血糖仪检测大鼠空腹血糖;HE染色检测大鼠视网膜组织病理结构;Western blot检测大鼠视网膜组织及rRMECs中的HIF-1α与血管内皮生长因子(VEGF)蛋白水平;RT-qPCR检测大鼠视网膜组织及rRMECs中的HIF-1α与VEGF mRNA水平;EdU试剂盒染色检测rRMECs增殖能力;对应试剂盒检测总胆固醇(TC)、三酰甘油(TG)、低密度胆固醇(LDL-C)、白细胞介素-6(IL-6)及肿瘤坏死因子-α(TNF-α)含量;CCK-8实验检测rRMECs细胞活性;Trallswell检测rRMECs细胞侵袭能力;免疫荧光染色检测HIF-1α与VEGF蛋白的表达水平。结果 与DR组相比,芍药苷组大鼠空腹血糖、TC、TG及LDL-C含量均减少,差异均有统计学意义(均为P<0.05)。与对照组相比,DR组大鼠视网膜组织疏松,界限不清;与DR组相比,芍药苷组大鼠视网膜组织疏松程度改善,界限略清。与对照组相比,DR组大鼠视网膜组织中的HIF-1α与VEGF的蛋白及mRNA表达水平,以及TNF-α与IL-6含量均增加,差异均有统计学意义(均为P<0.05)。与DR组相比,芍药苷组大鼠视网膜组织中的HIF-1α与VEGF的蛋白及mRNA表达水平,以及TNF-α与IL-6含量均减少,差异均有统计学意义(均为P<0.05)。与对照组相比,高糖组rRMECs的细胞活性、细胞增殖及侵袭能力均增加,差异均有统计学意义(均为P<0.05);与高糖组相比,芍药苷组rRMECs的细胞活性、细胞增殖及侵袭能力均降低,差异均有统计学意义(均为Objective To investigate the mechanisms by which paeoniflorin improves tissue and cell damage caused by diabetic retinopathy(DR) through the hypoxia-induced factor-1α(HIF-1α) pathway.Methods Thirty rats were randomly divided into a control group(10 normal rats),a DR group(10 diabetic model rats) and a paeoniflorin group(10 diabetic model rats given 80 mg·kg~(-1) paeoniflorin by gavage).Rat retinal microvascular endothelial cells(rRMECs) were divided into a control group(cultured with 5 mmol·L~(-1) glucose),a high glucose group(cultured with 30 mmol·L~(-1) glucose) and a paeoniflorin group(cultured with 30 mmol·L~(-1) glucose and 20 mol·L~(-1) paeoniflorin).The three groups of cells were all cultured for 24 h.Fasting blood glucose was measured by a glucose meter.Hematoxylin and eosin(HE) staining was used to detect the retinal histopathological structure.The levels of HIF-1α and vascular endothelial growth factor(VEGF) proteins and mRNAs in retinal tissues and rRMECs were detected by Western blotting and real time quantitative polymerase chain reaction(RT-qPCR).The proliferative ability of rRMECs was detected by the EdU kit.The serum levels of total cholesterol(TC),triglyceride(TG),low density cholesterol(LDL-C),interleukin-6(IL-6) and tumor necrosis factor-α(TNF-α) in retinal tissues and rRMECs were detected by kits.The activity and invasive ability of rRMECs were measured by CCK-8 and Transwell assay,respectively.The levels of HIF-1α and VEGF proteins in rRMECs were detected by immunofluorescence staining.Results Compared with those in the DR group,the fasting blood glucose,TC,TG and LDL-C levels in the paeoniflorin group were significantly decreased(all P<0.05).The retinal tissue was loose with an unclear boundary in the DR group,compared with that in the control group.The retinal tissue in the paeoniflorin group was less loose with a clearer boundary than that in the DR group.The levels of HIF-1α and VEGF proteins and mRNAs,TNF-α and IL-6 in retinal tissues of the DR group were significantly highe

关 键 词：糖尿病视网膜病变 缺氧诱导因子-1α通路 芍药苷 视网膜微血管内皮细胞 炎性因子 

分 类 号：R774.1[医药卫生—眼科]