摘要
目的从鳖甲临床抗肝纤维化功效出发并结合近红外光谱,探索建立关联抗肝纤维化活性的鳖甲质量生物活性评价方法。方法建立转化生长因子-β1(transforming growth factor-β1,TGF-β1)诱导的LX-2肝纤维化细胞模型,采用qRTPCR检测鳖甲对I型胶原(type I collagen,COL1)m RNA表达的影响,计算鳖甲对COL1 m RNA相对表达量的抑制率;采用Western blotting检测鳖甲对COL1蛋白表达的影响,以评价鳖甲在细胞模型上的抗肝纤维化生物活性。采集并处理不同批次鳖甲药材的近红外光谱数据,通过谱-效相关分析探究鳖甲药材近红外光谱数据与其抗肝纤维化活性的相关性,筛选活性波段,建立鳖甲的效应近红外谱,以效应近红外谱下面积评价鳖甲药材的质量。结果体外实验表明,与模型组比较,鳖甲水提物(water extract of Trionycis Carapax,WECT)处理组细胞中COL1 mRNA与蛋白表达水平显著下调(P<0.05、0.01);基于谱-效相关分析,共筛选得到R>0.6、P<0.05且波数大于8 cm^(-1)的活性波段3个,分别为5226~5235 cm^(-1)、5943~6071cm^(-1)与6379~6396 cm^(-1),以3个活性波段下面积为指标,建立鳖甲药材的效应近红外谱,计算各特征波段下面积和,并分析其与生物活性的相关性,两者相关系数R为0.81353。结论WECT可抑制LX-2细胞外基质的产生来治疗肝纤维化;效应波谱下面积和(5226~5235 cm^(-1)、5943~6071 cm^(-1)与6379~6396 cm^(-1))作为鳖甲药材的质量评价系数,用于评价鳖甲药材的质量,可区分不同批次鳖甲药材的抗肝纤维化活性。
Objective To explore the biological activity evaluation method of Biejia(Trionycis Carapax)quality related to anti-hepatic fibrosis activity based on its clinical anti-hepatic fibrosis effect and combined with near-infrared spectroscopy.Methods Transforming growth factor-β1(TGF-β1)induced LX-2 liver fibrosis cell model was established,qRT-PCR was used to detect the effect of Trionycis Carapax on type I collagen(COL1)mRNA expression,the inhibition rate of COL1 mRNA expression was calculated;Western blotting was used to detect the effect of Trionycis Carapax on COL1 protein expression to evaluate the anti-hepatic fibrosis bioactivity of Trionycis Carapax on cell model.Near-infrared spectral data of different batches of Trionycis Carapax were collected and processed,the correlation between near-infrared spectral data of Trionycis Carapax and their anti-liver fibrosis activity through spectrum effect correlation analysis was explored,and the active band was screened.The effect-near infrared spectrum of Trionycis Carapax was established,and the area under the effect near infrared spectrum was used to evaluate the quality of Trionycis Carapax.Results In vitro experiments showed that compared with model group,COL1 mRNA and protein expressions in water extract of Trionycis Carapax(WECT)were significantly decreased(P<0.05,0.01).Based on spectrum effect correlation analysis,three active bands with R>0.6,P<0.05 and wave number greater than 8 cm^(−1) were screened,which were 5226—5235 cm^(−1),5943—6071 cm^(−1) and 6379—6396 cm^(−1),respectively.Taking the area under the three active wavebands as the index,the effective nearinfrared spectrum of Trionycis Carapax was established,the area sum under each characteristic waveband was calculated,and its correlation with biological activity was analyzed.The correlation coefficient between the two was R=0.81353.Conclusion WECT can inhibit the production of LX-2 extracellular matrix to treat liver fibrosis;The sum of the area under the effect spectrum(5226—5235 cm^(−1),5943—6071 cm^(−1) and 6379—6396 cm^(−1))is used as the quality evaluation coefficient of Trionycis Carapax to evaluate the quality of Trionycis Carapax,which can distinguish the anti-liver fibrosis activity of different batches of Trionycis Carapax medicine.
作者
曹梦珍
黄倩
牛明
赵旭
肖小河
CAO Meng-zhen;HUANG Qian;NIU Ming;ZHAO Xu;XIAO Xiao-he(College of Pharmacy,Chengdu University of Traditional Chinese Medicine,Chengdu 610032,China;Department of Hepatology,the Fifth Medical Center of Chinese PLA General Hospital,Beijing 100039,China)
出处
《中草药》
CAS
CSCD
北大核心
2023年第10期3150-3157,共8页
Chinese Traditional and Herbal Drugs
基金
国家自然科学基金重点项目(81930110)
国家中医药传承创新团队项目(ZYYCXTD-C-202005)。
关键词
鳖甲
肝纤维化
质量评价
生物评价
近红外光谱
谱效相关分析
Trionycis Carapax
liver fibrosis
quality evaluation
biological evaluation
near-infrared spectroscopy
spectrum-effect correlation analysis
作者简介
曹梦珍(1997-),女,硕士研究生。E-mail:Caomengzhen1213@163.com;通信作者:肖小河,主要从事中药鉴定学和临床中药学工作。Tel:(010)66933322,E-mail:pharmacy302xxh@126.com;通信作者:赵旭,主要从事天然药物化学研究。E-mail:xuzhao080@outlook.com。
