基于均匀设计的腐尽生肌散有效单体成分配伍对HUVEC血管新生的影响  

Effect of Compatibility of Effective Monomer Components of Fujin Shengjisan on Angiogenesis of HUVEC Based on Uniform Design

作　　者：卢贤英 高静[1] 柏丁兮[1] 侯朝铭[1] 纪雯婷 陈欢 吴晨曦[1] LU Xianying;GAO Jing;BAI Dingxi;HOU Chaoming;JI Wenting;CHEN Huan;WU Chenxi(School of Nursing,Chengdu University of Traditional Chinese Medicine,Chengdu 610075,China)

机构地区：[1]成都中医药大学护理学院,成都610075

出　　处：《中国实验方剂学杂志》2025年第8期9-20,共12页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金青年基金项目(82104877);成都中医药大学杏林学者项目(MPRC2021021);护理学四川省重点实验室资助项目(HLKF2022-1);四川省基层卫生事业发展中心科研项目(SWFZ23-Y-23)。

摘　　要：目的:通过均匀设计,确定腐尽生肌散中有效单体成分“槲皮素-山柰酚-松香酸-乳香酸”配伍促糖尿病溃疡(DU)创面愈合的最佳配伍组合,实现古方腐尽生肌散现代化应用。方法:遵循“均匀设计-药效试验-数学建模及模型验证”的思路,按照U14(14~5)均匀设计方案,以4味中药单体成分为考察因素,葡萄糖诱导的人脐静脉内皮细胞(HUVEC)的增殖率为药效学指标,使用DPS软件构建有效单体成分与药效学指标间的数学模型,最后通过细胞增殖与活性检测法(CCK-8)、细胞划痕愈合、小管形成、蛋白免疫印迹法(Western blot)和实时荧光定量聚合酶链式反应(Real-time PCR)对均匀设计结果进行验证。结果:在14组配伍中,与高糖模型组比较,配伍6组差异具有统计学意义且促增殖效应最强(P<0.05);经DPS建模后最终拟合出4个二次多项式回归方程Y_(1)~Y_(4),考虑到模型的拟合度、稳定性及实际应用的需要,方程Y_(1)~Y_(3)用于后续验证;同时为确保实验重复性,配伍6组也用于验证;并依次将配伍6组、方程Y_(1)~Y_(3)组重命名为新复方(1)~新复方(4),作为古方腐尽生肌散进行现代化应用的单体成分配伍形式。验证实验结果表明:CCK-8、划痕愈合和小管形成实验均表明,与空白组比较,50 mmol·L^(-1)葡萄糖刺激HUVEC后,其相应的细胞活力、创面愈合率和成管数量均明显下降;此外,还明显下调了血管生成相关细胞因子血管内皮生长因子(VEGF)和成纤维细胞生长因子2(FGF2)的表达水平及CD31的分泌。而给予复方(1)~(4)对其干预后,其中复方(1)和复方(3)则显著改善了50 mmol·L^(-1)葡萄糖诱导HUVEC损伤后的这些生物学功能。进一步从复方(1)和复方(3)的回归系数及各单体成分的相对剂量占比推断,松香酸、槲皮素、乳香酸均有促高糖环境下HUVEC血管新生的作用,且发挥主效应(偏相关系数为正,均>0.9);松香酸和乳香酸、山柰酚和乳香�Objective:To determine the optimal combination of the effective monomer components"quercetin-kaempferol abietic acid-boswellic acid"in Fujin Shengjisan for promoting diabetic ulcer(DU)wound healing through uniform design,thereby achieving the modern application of the ancient formula.Methods:Following the principle of"uniform design-pharmacodynamic experiment-mathematical modeling and model verification",the U14(145)uniform design table was adopted.The four monomer components of Chinese medicine were considered as the independent variables,and the proliferation rate of human umbilical vein endothelial cells(HUVECs)induced by glucose was used as the pharmacodynamic indicator.A mathematical model was constructed using DPS software to correlate the effective monomer components with the pharmacodynamic indicator.The results of uniform design were verified through CCK-8 assay,cell scratch healing,tube formation,Western blot,and Real-time PCR.Results:Among the 14 compatibility groups,compared with the high-glucose model group,compound compatibility group 6 showed the strongest proliferation effect and statistical significance(P<0.05).Four quadratic polynomial regression equations(Y_(1)Y_(4))were obtained through DPS modeling.Considering the model's fit,stability,and practical application,equations Y_(1)-Y_(3)were selected for the follow-up verification.To ensure experiment reproducibility,group 6 was used for validation.Group 6 and equations Y_(1)-Y_(3)were renamed as compound prescription①to compound prescription④,respectively,to represent the modern application of the ancient FJSJ Powder through compatibility of monomer components.Verification experiments showed that in the CCK-8,scratch healing,and tube formation assays,the cell viability,wound healing rate,and tube formation number of HUVECs stimulated with 50 mmol·L^(-1)glucose were significantly reduced compared with the blank group.Moreover,the expression levels of angiogenesis-related cytokines,vascular endothelial growth factor(VEGF)and fibroblast growth

关 键 词：腐尽生肌散 糖尿病溃疡 血管新生 均匀设计 

分 类 号：R285[医药卫生—中药学] R587.1[医药卫生—中医学] R543