唐古特大黄FTIR的双指标序列分析及抗炎谱效关系研究

Double Index Sequence Analysis of FTIR and Anti-Inflammatory Spectrum Effect Relationship of Rheum Tanguticum

对于中药材的质量控制研究是中医药现代化的主要研究内容之一。唐古特大黄是一种常用中药,目前对唐古特大黄的质量控制研究多集中在总蒽醌类成分、游离蒽醌类成分以及指纹图谱,基于抗炎效果的质量标准研究尚为少数。采集不同产地、不同采收时间的18批唐古特大黄药材,对其粉末进行傅里叶变换红外光谱(FTIR)研究,发现16个共有峰,18批次之间具有较高的相似性(0.798~0.900),相同产地(甘南产、青海产)的唐古特大黄相似度较高(相似度大于0.900的达80%以上),但两产地之间的相似度相对较低(相似度大于0.900的仅为69.44%),结合聚类分析发现不同产地唐古特大黄FTIR谱图存在一定差异性,产地可能是主要影响因素。建立双指标分析序列,可在一定程度上区分药材不同产地。脂多糖(LPS)刺激RAW264.7细胞建立炎性细胞模型,以NO抑制率作为炎性指标,测定不同批次唐古特大黄的NO抑制率。以共有峰透过率作为X, NO抑制率作为Y,采用SMICA14.1软件构建PLSR模型建立谱效关系,谱图中X_(14)、X_(15)、X_(11)、X_(2)四个共有峰VIP>1,其中X_(2)与抗炎效果呈正相关关系,X_(14)、X_(15)、X_(11)与抗炎效果现负相关关系。进一步建立数学模型得共有峰(X)与NO抑制率(Y)的关系Y=237.618+2.992X_(2)-0.845X_(4)+2.979X_(6)-3.722X_(7)+0.433X_(8)-0.957X_(12)-0.759X_(14)-0.632X_(15)(p=0.003<0.01,R=0.935),p<0.01代表构建数学模型有意义,R=0.935代表所构建数学模型可解释93.5%的数据。可知X_(2)、X_(6)与抗炎效果呈正相关关系,X_(4)、X_(7)与抗炎效果呈负相关关系,可预测体外抗炎效果,准确率达93.5%,但由于样本量过小,仅可用于解释本研究纳入数据。X_(6)是由甲基的β(C—H)产生,可能与大黄素、大黄酸、大黄素甲醚、芦荟大黄素等蒽醌类成分有关。该研究将抗炎活性与红外光谱数据相结合,引入唐古特大黄质量评价体系,为唐古特大黄产业的健康发展提供新思路。

Research on the quality control of Chinese medicinal materials is one of the main aspects of the modernization of traditional Chinese medicine.Rheum taanduticum(R.tanguticum)is one of the most widely used Chinese medicinal materials.The research on the quality control of R.tanguticum is total anthraquinones,anthraquinones,fingerprint and so on.There are few studies on quality standards based on anti-inflammatory effect in the study of R.tanguticum.Eighteen batches of R.tanguticum from different origins and processing conditions were collected to study by Fourier-transform infrared spectroscopy(FTIR).FTIR founded sixteen common peeks.There is high similarity(0.798~0.900)in 18 batches of R.tanguticum.There is a high similarity(the similarity greater than 0.900 is more than 80%)in the same origin(Gannan,Qinghai).The similarity of R.tanguticum in the same origin(Gannan,Qinghai)was higher(more than 80%if the similarity is greater than 0.900)than that between the two origins(only 69.44%if the similarity is greater than 0.900).There were variations in FTIR spectra of R.tanduticum in different origins,combining the result of cluster analysis.The origins maybe the main influence factor in the FTIR spectra.Furthermore,the sequence of double index analysis was established to distinguish different origins of medicinal materials.LPS stimulated RAW 264.7 cells to establish an inflammatory cell model.NO inhibition rate was used as an inflammatory index.Determination of NO inhibition rate of 18 batches of R.tanduticum.Establishing the PLSR model with common peaks(X)and NO inhibition rate(Y)by SMICA 14.1 software.There were 4 common peaks(X_(14),X_(15),X_(11),X_(2)),which VIP>1.It was found that X_(2) in the FTIR spectrum was positively correlated with an anti-inflammatory effect,and X_(14),X_(15),X_(11) were negatively correlated with anti-inflammatory effect.By further establishing the mathematical model with common peaks(X)and NO inhibition rate(Y),Y=237.618+2.992X_(2)-0.845X_(4)+2.979X_(6)-3.722X_(7)+0.433X_(8)-0.957X_(12)-0.759X_(14)-0.632X_(15)(p=0.003<0.01,R=0.935),it can be seen that X_(2),X_(6) have a positive correlation with the anti-inflammatory effect,and X_(4),X_(7) have a negative correlation with the anti-inflammatory effect.The in vitro anti-inflammatory effect can be predicted by the FTIR spectrum of R.tanguticum,with an accuracy of 93.50%.However,due to the small sample size,it can only be used to explain the data included in this study.X_(6) is made of methylβ(C—H),which may be related to the content of anthraquinone components(such as Emodin,Rhein,Physcion,Aloe emodin).The results of this study can introduce the anti-inflammatory efficacy evaluation into the quality evaluation system of Rheum tanguticum,enrich the quality evaluation system of R.tanguticum,and provide new ideas for the healthy development of Rheum tanguticum industry.