条芩与枯芩的指纹图谱建立、抗炎活性及谱效关系研究
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篇名: 条芩与枯芩的指纹图谱建立、抗炎活性及谱效关系研究
TITLE: Establishment of fingerprints ,anti-inflammatory activities and spectrum-effect relationship study of pith-nodecayed and pith-decayed Scutellaria baicalensis
摘要: 目的 建立条芩与枯芩的指纹图谱,考察两者的抗炎活性差异。方法采用高效液相色谱法建立5批条芩和5批枯芩的指纹图谱;采用SIMCA14.1软件,以变量重要性投影(VIP)值>1为标准筛选影响条芩和枯芩质量的差异标志物;以其对脂多糖(LPS)诱导的小鼠单核巨噬细胞(RAW264.7细胞)上清液中一氧化氮(NO)、白细胞介素6(IL-6)、IL-1β的抑制率为指标,评价条芩和枯芩的抗炎活性并计算其抑制NO的半数有效浓度(EC50);采用灰色关联分析谱效关系;以50μg/mL质量浓度下各组细胞的NO、IL-1β、IL-6抑制率为指标,采用SPSS26.0软件进行聚类分析。结果5批条芩和5批枯芩共有15个共有峰,相似度均大于0.990。共有8个峰的VIP值>1,依次为峰14、12、15、6、10、13、11、4。在50μg/mL质量浓度下,枯芩提取物对NO、IL-6、IL-1β的抑制率及平均EC50分别为62.14%~71.13%、3.32%~18.38%、93.12%~95.47%、25.35μg/mL,条芩提取物分别为39.52%~50.19%、6.21%~22.55%、94.10%~96.44%、58.63μg/mL;枯芩提取物的平均NO抑制率显著高于条芩提取物,平均EC50、IL-6抑制率均显著低于条芩提取物(P<0.05);两者平均IL-1β抑制率比较差异无统计学意义(P>0.05)。峰2~3、5~8、10~11与NO抑制率的关联度均大于0.8,峰2、5、8~9与IL-1β抑制率的关联度均大于0.9,15个共有峰与IL-6抑制率的关联度均小于0.8。10批样品可聚为2类,K1~K5为一类,T12~T16为一类。结论在50µg/mL质量浓度下,枯芩提取物对NO的抑制作用强于条芩提取物,对IL-6的抑制作用弱于条芩提取物,对IL-1β的抑制作用与条芩提取物相当;初步确定峰6、10~11对应成分为条芩与枯芩抗炎活性差异的主要化学成分。
ABSTRACT: OBJECTIVE To es tablish the fingerprints of pith-nodecayed and pith-decayed Scutellaria baicalensis ,and to investigate the difference of their anti-inflammatory activities. METHODS HPLC method was adopted to establish the fingerprints of 5 batches of pith-nodecayed S. baicalensis and 5 batches of pith-decayed S. baicalensis . SIMCA 14.1 software was adopted to screen the markers of weight difference between pith-decayed S. baicalensis and pith-nodecayed S. baicalensis ,using variable importance in projection (VIP)>1 as standard. Using the inhibitory rates of NO ,IL-6 and IL- 1β in supernatant of lipopo- lysaccharide(LPS)-induced monocyte macrophages (RAW264.7 cells)as indexes ,the anti-inflammatory activities of pith-decayed and pith-nodecayed S. baicalensis were evaluated and median effective concentration (EC50)of NO were calculated. The gray correlation method was used for spectrum-effect relationship analysis ;SPSS 26.0 software was used for cluster analysis using the inhibitory rates of NO ,IL-6 and IL- 1β in cells at the concentration of 50 μg/mL. RESULTS There were 15 common peaks in 5 batches of pith-nodecayed S. baicalensis and 5 batches of pith-decayed S. baicalensis ,and the similarity of them was greater than 0.990. There were total of 8 peaks with VIP 1.0,they were peak 14,12,15,6,10,13,11 and 4. At jcqinge@163.com the concentration of 50 μg/mL,the inhibitory rates of pith- decayed S. baicalensis extract to NO ,IL-6,IL-1 β and the average EC 50 were 62.14%-71.13%,3.32% -18.38% and 93.12%-95.47% and 25.35 μg/mL,respectively;those of pith-nodecayed S. baicalensis extract were 39.52%-50.19%, 6.21%-22.55%,94.10%-96.44% and 58.63 μg/mL,respectively. Average NO inhibition rate of pith-decayed S. baicalensis extract was significantly higher than that of pith- nodecayed S. baicalensis extract;average EC 50 and average IL- 6 inhibitory rate of pith-decayed S. baicalensis extract were significantly lower than those of pith-nodecayed S. baicalensis extract(P<0.05). There was no statistical significance in average inhibitory rate of IL- 1β(P>0.05). The correlation degrees of peaks 2-3,5-8 and 10-11 with inhibitory rate of NO were all greater than 0.8. The correlation degrees of peaks 2,5,8-9 with inhibitory rate of IL- 1β were all greater than 0.9. The correlation degrees of 15 common peaks with inhibitory rate of IL- 6 were all less than 0.8. Ten batches of samples could be grouped into two categories ,K1-K5 were clustered into one category and T 12-T16 were clustered into one category. CONCLUSIONS At the concentration of 50 µg/mL,the inhibitory effect of pith-decayed S. baicalensis extract to NO is stronger than that of pith-nodecayed S. baicalensis extract,while the inhibitory effect on IL- 6 is weaker than that of pith-nodecayed S. baicalensis extract;they have similar inhibitory effect on IL- 1β. The corresponding components of peak 6,10-11 are preliminarily identified as main chemical components of the difference in anti-inflammatory activity between pith-decayed S. baicalensis and pith-nodecayed S. baicalensis .
期刊: 2022年第33卷第12期
作者: 贾传青,郭兰萍,王晓,于宗渊,陈龙,董红敬
AUTHORS: JIA Chuanqing,GUO Lanping,WANG Xiao,YU Zongyuan,CHEN Long,DONG Hongjing
关键字: 黄芩;条芩提取物;枯芩提取物;指纹图谱;抗炎活性;谱效关系
KEYWORDS: Scutellaria baicalensis ; pith-nodecayed Scutellaria baicalensis extract; pith-decayed Scutellaria baicalensis
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