紫朱醇提取物对高糖高脂诱导血管内皮细胞损伤的保护作用  

Protective effect of ethanol extract of Zizhu on vascular endothelial cell injury induced by high glucose and high lipids

作　　者：王莉娟 柳国斌 韩秋琴 赵志弘 何金晶 李文惠 Wang Lijuan;Liu Guobin;Han Qiuqin;Zhao Zhihong;He Jinjing;Li Wenhui(Shanghai University of Medicine&Health Sciences,Shanghai 201318,China;Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China)

机构地区：[1]上海健康医学院,上海市201318 [2]上海中医药大学,上海市201203

出　　处：《中国组织工程研究》2023年第26期4132-4138,共7页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金青年项目(81804096),项目负责人:李文惠;上海健康医学院师资人才百人库国内访学项目(A3-0200-21-311007-12),项目负责人:李文惠。

摘　　要：背景:紫朱软膏由黄芪、紫草、朱砂、龙血竭、阿胶、冰片6味中药组成,有清热解毒、祛腐生肌、补气益血的功效。前期临床和动物实验已证实其在糖尿病溃疡愈合过程中发挥抑制炎症、促进血管生成的作用。目的:在细胞水平上探究紫朱醇提取物对高糖高脂诱导小鼠脑微血管内皮细胞(bEnd.3细胞)损伤的保护作用,并研究其对血管生成的影响。方法:模拟高糖高脂环境,采用不同质量浓度的葡萄糖和棕榈酸诱导bEnd.3细胞损伤,给予不同质量浓度的紫朱醇提取物干预,用CCK-8法筛选出最合适的刺激物质量浓度和最佳给药质量浓度;然后通过划痕实验、血管生成实验观察紫朱醇提取物对bEnd.3细胞促血管生成的影响,采用AnnexinV/PI凋亡试剂盒检测bEnd.3细胞凋亡率,采用Western blot和qRT-PCR法检测bEnd.3细胞中VEGFA、Ang-2、SPRED1、PIK3R2的蛋白和mRNA表达水平。结果与结论:①经CCK-8法检测细胞活力,最终选定25 mmol/L葡萄糖+200μmol/L棕榈酸作用24 h作为模型组,500 mg/L紫朱醇提取物作用24 h作为紫朱醇提取物组给药方式;②与正常对照组相比,模型组bEnd.3细胞迁移和生成血管能力下降(P<0.001,P<0.05),bEnd.3细胞凋亡率上升(P<0.001),VEGFA、Ang-2 mRNA和蛋白表达显著降低(P<0.001),SPRED1、PIK3R2 mRNA和蛋白表达显著增加(P<0.001);③与模型组相比,紫朱醇提取物组bEnd.3细胞迁移和血管生成能力上升(P<0.05),bEnd.3细胞凋亡率显著下降(P<0.001),VEGFA、Ang-2 mRNA和蛋白表达显著增加(P<0.001),SPRED1、PIK3R2 mRNA和蛋白表达显著降低(P<0.001,P<0.05);④结果表明,紫朱醇提取物对高糖高脂诱导的bEnd.3细胞具有保护作用,能够促进细胞迁移和血管生成并减少细胞凋亡。BACKGROUND:Zizhu ointment is composed of Astragali Radix,Radix Arnebiae seu Lithospermi,Cinnabar,Resina Dracoins,Ejiao and Borneol,which has the effects of clearing heat and detoxifying,removing saprophyxia and strengthening muscles,invigorating qi and benefiting blood.Previous clinical and animal experiments have shown that it can inhibit inflammation and promote angiogenesis in the healing process of diabetic ulcer.OBJECTIVE:To investigate the protective effect of ethanol extract of Zizhu on the injury of rat brain microvascular endothelial cells(bEend.3 cells)induced by high glucose and high lipids at the cellular level,and to study its effect on angiogenesis.METHODS:bEnd.3 cells were induced with glucose and palmitic acid at different concentrations,and treated with different concentrations of ethanol extract of Zizhu.The optimum concentrations for modeling and administration were identified by cell counting kit-8 method.The effect of ethanol extract of Zizhu on angiogenesis of bEnd.3 cells was observed by scratch test and angiogenesis test.AnnexinV/PI apoptosis detection kit was used to detect the apoptosis rate of bEnd.3 cells under fluorescence microscope.The protein and mRNA expression levels of VEGFA,Ang-2,SPRED1,and PIK3R2 in bEnd.3 cells were determined by western blot and qRT-PCR,respectively.RESULTS AND CONCLUSION:Cell viability was detected by cell counting kit-8 method.25 mmol/L glucose+200μmol/L palmitic acid for 24 hours was selected as the model group;treatment with ethanol extract of Zizhu 500 mg/L for 24 hours was performed as the way of administration in the ethanol extract of Zizhu group.Compared with the normal group,bEnd.3 cell migration and angiogenesis were decreased in the model group(P<0.001,P<0.05),bEnd.3 cell apoptosis rate was increased(P<0.001),the mRNA and protein expressions of VEGFA and Ang-2 were significantly decreased(P<0.001),and the mRNA and protein expressions of SPRED1 and PIK3R2 were significantly increased(P<0.001).Compared with the model group,in the ethanol extract o

关 键 词：紫朱醇提取物 高糖高脂 小鼠脑微血管内皮细胞 血管生成 血管内皮生长因子 

分 类 号：R459.9[医药卫生—治疗学] R318[医药卫生—临床医学] R587.1