冻干与生晒西洋参饮片差异皂苷含量及促斑马鱼血管生成活性比较  

Comparison of Differential Saponin Contents and Pro-angiogenic Activities on Zebrafish Between Freeze-dried and Sun-dried Panacis Quinquefolii Radix Decoction Pieces

作　　者：邵娟 徐梦丹 张雪晶 郭云龙 蔡广知[1] 贡济宇[1] SHAO Juan;XU Mengdan;ZHANG Xuejing;GUO Yunlong;CAI Guangzhi;GONG Jiyu(School of Pharmaceutical Sciences,Changchun University of Chinese Medicine,Changchun 130117,China;Jilin Ginseng Academy,Changchun University of Chinese Medicine,Changchun 130117,China)

机构地区：[1]长春中医药大学药学院,长春130117 [2]长春中医药大学吉林省人参科学研究院,长春130117

出　　处：《中国实验方剂学杂志》2024年第17期185-192,共8页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：吉林省科技发展计划项目(20240305014YY)。

摘　　要：目的:分析和测定冻干与生晒西洋参饮片中的差异成分,并比较二者促血管生成活性的差异。方法:基于超高效液相色谱法(UPLC)建立冻干和生晒西洋参饮片的指纹图谱,结合主成分分析(PCA)、正交偏最小二乘法-判别分析(OPLSDA)等化学计量学方法确定2种饮片的差异皂苷成分,采用UPLC测定其中6种代表性成分在冻干与生晒西洋参饮片中的含量;选取受精24 h的转基因斑马鱼品系Tg(fli1a∶EGFP)胚胎,分别使用不同剂量的冻干与生晒西洋参70%甲醇提取物(10、30 mg·L^(-1))干预正常斑马鱼和血管内皮生长因子受体酪氨酸激酶抑制剂Ⅱ(PTK787)诱导的节间血管(ISV)损伤斑马鱼模型,观察斑马鱼肠下静脉(SIV)和ISV发育情况,测定SIV直径、平均交叉数和平均发芽数,计算ISV血管指数,比较2种饮片的促血管生成活性。结果:冻干与生晒西洋参饮片指纹图谱相似度均>0.950,分别确定了17个共有峰,指认了其中6个共有峰分别为峰6(人参皂苷Rg_(1))、峰7(人参皂苷Re)、峰8(人参皂苷Rb_(1))、峰11(人参皂苷Rc)、峰13(人参皂苷Rb_(2))、峰16(人参皂苷Rd);PCA及OPLS-DA筛选出11种差异性皂苷成分,表明2种饮片的成分含量存在一定差异;含量测定结果显示,冻干西洋参饮片中人参皂苷Rg_(1)、Re、Rb_(1)、Rb_(2)含量高于生晒西洋参饮片,而人参皂苷Rc、Rd含量低于生晒西洋参饮片(P<0.05,P<0.01)。在对正常斑马鱼胚胎促血管生成作用研究中,与空白组比较,冻干及生晒西洋参高剂量组SIV血管直径、平均发芽率及平均交叉率均显著升高(P<0.01),且冻干西洋参组血管直径、平均交叉数及平均发芽数均高于生晒西洋参组(P<0.05)。对ISV损伤的斑马鱼胚胎促血管生成作用研究显示,与空白组比较,模型组斑马鱼ISV发育受到抑制;与模型组比较,冻干及生晒西洋参各剂量组均可促进ISV的生长与萌发,相同剂量下,冻干西洋参组的正常血管数目明显�Objective:To analyze and determine the differential components of freeze-dried and sundried Panacis Quinquefolii Radix(PQR),and to compare the differences in their pro-angiogenic activities.Method:Fingerprints of freeze-dried and sun-dried PQR were established based on ultra performance liquid chromatography(UPLC),and chemometrics methods such as principal component analysis(PCA)and orthogonal partial least squares-discriminant analysis(OPLS-DA)were combined to determine the differential saponin composition of the two decoction pieces,and six representative saponins were selected and their contents in freeze-dried and sun-dried PQR were determined by UPLC.Transgenic zebrafish line Tg(fli1a∶EGFP)embryos fertilized for 24 h were selected,and different doses of 70%methanol extracts of freeze-dried and sun-dried PQR(10,30 mg·L^(-1))were used to intervene in normal zebrafish and in a zebrafish model of intersegmental vascular(ISV)injury induced by vascular endothelial growth factor(VEGF)receptor tyrosine kinase inhibitorⅡ(PTK787),then the development of subintestinal vein(SIV)and ISV of zebrafish was observed,SIV diameter,mean number of crossings and mean number of germinations were determined,and the ISV vascular index was calculated,in order to compare the pro-angiogenic activities of the two decoction pieces.Result:The similarity of the fingerprints of freeze-dried and sun-dried PQR decoction pieces was>0.950,and 17 common peaks were identified,of which 6 common peaks were designated as peak 6(ginsenoside Rg_(1)),peak 7(ginsenoside Re),peak 8(ginsenoside Rb_(1)),peak 11(ginsenoside Rc),peak 13(ginsenoside Rb_(2)),and peak 16(ginsenoside Rd),respectively.A total of 11 differential saponin components were screened by PCA and OPLS-DA,indicating that there were some differences in the contents of the components in the two decoction pieces.The results of determination showed that the contents of ginsenosides Rg_(1),Re,Rb_(1) and Rb_(2) in freezedried PQR were higher than those in sun-dried PQR,while the contents of

关 键 词：西洋参饮片 冻干 生晒 指纹图谱 含量测定 化学计量法 促血管生成 

分 类 号：R22[医药卫生—中医基础理论] R825.4[医药卫生—中医学] R28R943.1