摘要
目的观察丹酚酸B对高糖培养的视网膜内皮细胞迁移及管道形成的影响,并采用网络药理学方法探讨其作用机制。方法将细胞按随机数字表法分为正常组及1.0、0.5、0.1 μg/ml丹酚酸B组。各组细胞加入5.5 mmol/L葡萄糖进行干预,1.0、0.5、0.1 μg/ml丹酚酸B组分别加入1.0、0.5、0.1 μg/ml丹酚酸B进行干预。72 h后,采用CCK-8法检测各组细胞活力。将细胞按随机数字表法分为正常组、模型组及丹酚酸B低、中、高剂量组。正常组细胞加入5.5 mmol/L葡萄糖进行干预。模型组加入25 mmol/L葡萄糖干预;丹酚酸B低、中、高剂量组分别加入25 mmol/L葡萄糖及0.062 5、0.125 0、0.250 0 μg/ml丹酚酸B进行干预。采用Transwell实验检测细胞迁移数目,Matrigel实验分析细胞成管总长度。通过SuperTarget与Swiss TargetPrediction筛选丹酚酸B的活性作用靶点;检索GAD数据库、pharmGkb数据库、TTD数据库、DiGSeE数据库和OMIM数据库,获取糖尿病视网膜病变的相关靶点;两者取交集得到共同靶点,通过Cytoscape构建成分-靶点-疾病相互作用网络;利用DAVID对共同靶点进行GO分析及KEGG通路富集分析;运用Accelrys Discovery Studio Client 2.5软件进行分子对接。结果 CCK-8法检测结果显示,0.5、0.1 μg/ml丹酚酸B组细胞吸光度值与正常组比较差异无统计学意义(P>0.05)。Transwell实验、Matrigel实验结果显示,与模型组比较,丹酚酸B各剂量组迁移细胞相对个数、细胞成管总长度减少(P<0.05或P<0.01)。通过网络药理学构建相互作用网络发现,丹酚酸B作用于46个靶点,8个信号通路。结论丹酚酸B可抑制高糖培养的视网膜血管内皮细胞的迁移能力和体外成管能力,并通过多靶点、多通路干预糖尿病视网膜病。
Objective To observe the effects of salvianolic acid B on migration and tube formation of the retinal vascular endothelial cell(RVEC)in high glucose,and explore its mechanism with network pharmacology.Methods The cells were divided into normal group,model group and 1.0,0.5,0.1μg/ml salvianolic acid B group according to the random number table method.The cells of each group were added with 5.5 mmol/L glucose for intervention,and the salvianolic acid B group was added with 1.0,0.5,and 0.1μg/ml salvianolic acid B for intervention.After 72 h,the cell viability of each group was detected by the CCK-8 method.The cells were divided into normal group,model group and low-,medium-,and high-dose salvianolic acid B group according to the random number table method.Then the cells of the normal group were added with 5.5 mmol/L glucose;the model group was added with 25 mmol/L glucose;the low-,medium-,and high-dose salvianolic acid B group was added with 25 mmol/L glucose and 0.0625,0.1250,0.2500μg/ml salvianolic acid B.Then by taking Transwell test to detect the number of cell migration,and Matrigel test to analyze the total length of cells tubes.The active targets of Salvianolic acid B were screened by SuperTarget and Swiss TargetPrediction.Then,the targets of diabetic retinopathy were obtained by searching the GAD database,pharmGkb database,TTD database,DiGSeE database and OMIM database.The effective targets of drug-disease interaction were screened,and the component-target-disease interaction network was constructed by Cytoscape.Finally,the effective targets were analyzed by DAVID for GO analysis and KEGG pathway enrichment analysis.Molecular docking was performed by using Accelrys Discovery Studio Client 2.5 software.Results The CCK-8 method showed that the cell absorbance values of 0.5 and 0.1μg/ml salvianolic acid B group were not significantly different from those of the normal group(P>0.05).The results of Transwell experiment and Matrigel experiment showed that compared with the model group,the relative number of mi
作者
蒋坤秀
孙惠惠
宋星卓
王析瑞
孙倩倩
田婧鋆
李宏丽
刘永刚
韩静
Jiang Kunxiu;Sun Huihui;Song Xingzhuo;Wang Xirui;Sun Qianqian;Tian Jingyun;Li Hongli;Liu Yonggang;Han Jing(School of Chinese Material Medica,Beijing University of Chinese Medicine,Beijing 100102,China;School of Traditional Chinese Medicine,Beijing University of Chinese Medicine,Beijing 100029,China;Institute of Traditional Chinese Medicine,Beijing University of Chinese Medicine,Beijing 100029,China)
出处
《国际中医中药杂志》
2020年第11期1094-1101,共8页
International Journal of Traditional Chinese Medicine
基金
国家自然科学基金(81673705、81873165)。
关键词
药理作用分子作用机制(中药)
丹酚酸B
网络药理学
视网膜
内皮细胞
Molecular mechanisms of pharmacological action(TCD)
Salvianolic acid B
Network pharmacology
Retina
Endothelial cells
