瑶药石柑子指纹图谱特征峰与其抗炎活性的关系研究

Research on relationship between characteristic peak of Herba Pothi Chinensis fingerprint and its anti-inflammatory activity

目的:研究分析石柑子指纹图谱特征峰与其抗炎作用的相关性,为精确评价石柑子的功效并研究其物质基础提供参考。方法:采用HPLC法,色谱条件为:以0.1%磷酸水溶液(A)-乙腈(B)为流动相;采用梯度洗脱,梯度程序为0~10min,10%~20%B;10~20min,20%~25%B;20~50min,25%~40%B;50~60min,40%~10%B;检测波长318nm,柱温25℃,流速1.0m L/min;细胞水平抗炎活性实验采用MTT法确定最大无毒浓度,进行不同提取部位干预下的RNA表达实验,根据表达结果确定抗炎活性。结果:石柑子三氯甲烷提取部位对RAW264.7细胞活性影响最为显著,不同提取部位指纹图谱中,峰2和峰5变化最为明显(以峰面积占比计,峰2为1.21%~8.65%,峰5为2.45%~5.13%),提示可能为石柑子抗炎活性的关键化合物。结论:该方法结合细胞实验和指纹图谱,能准确地找到石柑子抗炎功效的物质基础。

Objective：To investigate correlation between characteristic peaks in fingerprint and anti-inflammatory activity of Herba Pothi Chinensis,for more précised evaluation of its effectiveness and material basis; Methods：Analysis was carried out by HPLC,the mobile phase comprised 0.1% phosphoric acid（A）and acetonitrile（B）,using gradient as follow：0-10min,10%-20% B; 10-20min,20%-25% B; 20 -50min,25% -40% B; 50 -60min,40% -10% B; detection wave length：318nm,column temperature：25℃,flow rate：1.0m L/min anti-inflammatory experiment in bacterial level was carried out by MTT method,RAW264.7 cell was inoculated into a 96 well format for 48 h cultivation,followed by 24 h of stimulation by the herbal extract,then 5mg/m L of MTT was added and cultivated for4 h,the upper clear solvent was taken for absorbance determination; Results：trichloromethane fraction had strongest influences on bacteria vitality,among fingerprints of all fractions,peak 2 and 5 had greatest variation（for peak area ratio,the values were 1.21-8.65% for peak 2 and 2.45-5.13%,respectively）,indicating their key contribution to anti-inflammatory activity; Conclusion：the method integrated bacteria experiment and fingerprint was more précised in searching material basis of Herba Pothi Chinensis＇ anti-inflammatory activity.