生、醋柴胡UPLC-DAD指纹图谱建立及抗肝纤维化作用的谱效关系研究  

Establishment of UPLC-DAD fingerprint of raw and vinegar Bupleurum bupleurum and study on spectral effect relationship of anti-hepatic fibrosis

作　　者：陈妮平 王燕 董妍 肖扬鑫 涂济源 刘艳菊 周仲实 CHEN Ni-ping;WANG Yan;DONG Yan;XIAO Yang-xin;TU Ji-yuan;LIU Yan-ju;ZHOU Zhong-shi(School of Pharmacy,Hubei University of Chinese Medicine;Hubei Provincial Engineering Research Center for Traditional Chinese Medicine Processing,Wuhan 430065,China)

机构地区：[1]湖北中医药大学药学院 [2]湖北省中药炮制工程技术研究中心,湖北武汉430065

出　　处：《中国药理学通报》2024年第6期1145-1152,共8页Chinese Pharmacological Bulletin

基　　金：中药饮片药性及质量评价平台(No 2020ZYYD030)。

摘　　要：目的建立生、醋柴胡的指纹图谱,比较生、醋柴胡化学成分差异,考察两者抗肝纤维化作用的差异,探究其与抗肝纤维化药效之间的关联性。方法采用UPLC法建立10批生柴胡和10批醋柴胡的指纹图谱,以TGF-β诱导肝星状细胞(LX-2)建立肝纤维化体外细胞模型,以Ⅰ型胶原蛋白(collagenⅠ,col1a1)和α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)蛋白表达情况为药效指标,利用MetaboAnalyst 5.0网站,以变量重要性投影(VIP)值>1为标准筛选影响生、醋柴胡质量的差异标志物,并采用正交偏最小二乘判别分析(OPLS-DA)研究生、醋柴胡差异化学成分及与抗肝纤维化作用的谱效关系,筛选生、醋柴胡抗肝纤维化作用的主要成分。结果生、醋柴胡样品的UPLC指纹图谱共有峰有18个,生、醋柴胡的主要差异成分有9个,其中峰9、7、6可以指认为柴胡皂苷d、柴胡皂苷a、柴胡皂苷f。药效结果发现,生、醋柴胡均有抗纤维化活性,且醋柴胡抗纤维化活性更强。通过谱效关联结果发现,柴胡抗肝纤维化作用的主要相关成分为11、12、14、15、18号峰。结论建立生、醋柴胡的指纹图谱方法简单可行,通过谱效关系筛选出抗肝纤维化活性的重要成分,为明确生、醋柴胡抗肝纤维化作用物质基础提供依据。Aim To establish the fingerprint of raw bupleurum and vinegar bupleurum,investigate the difference in their anti-liver fibrosis effects,and explore the relationship between the chemical composition of raw bupleurum and vinegar bupleurum and their anti-liver fibrosis efficacy.Methods The fingerprints of 10 batches of raw bupleuri and 10 batches of bupleuri were established by UPLC method.The liver fibrosis cell model in vitro was established by TGF-βinduced LX-2 hepatic stellate cells.The liver fibrosis cell model was analyzed with collagen type I(col1a1)andα-smoothmuscleactin.The expression ofα-SMA protein was used as the pharmacodynamic index.MetaboAnalyst5.0 was used to screen the difference markers affecting the quality of raw bupledges and vinegar bupledges with VIP value>1 as the criterion.Orthogonal partial least squares discriminant analysis(OPLS-DA)was used to screen the main components of raw bupleurum and vinegar bupleurum against liver fibrosis.Results There were 18 peaks in the UPLC fingerprints of raw bupleurum and vinegar bupleurum,and the analysis results showed that there were nine main differences between raw bupleurum and vinegar bupleurum,among which peaks 9,7 and 6 could be considered as bupleurin d,bupleurin a and bupleurin f.The results of spectral effect relationship showed that the main components of bupleurum anti-liver fibrosis were peaks 11,12,14,15 and 18.Conclusions The established fingerprint method of raw bupleurum and vinegar bupleurum is simple and feasible,and the important components of anti-liver fibrosis activity are screened through the spectrum effect relationship,which provides a basis for clarifying the material basis of anti-liver fibrosis effect of raw bupleurum and vinegar bupleurum.

关 键 词：生柴胡 醋柴胡 指纹图谱 肝纤维化 正交偏最小二乘 谱效关系 

分 类 号：R284.1[医药卫生—中药学] R322.47[医药卫生—中医学] R575.2