桂龙凝胶膏剂抗风湿活性成分的筛选及质量控制研究

Gui Long gel paste anti-rheumatism screening of active ingredients,and quality control research

目的:观察桂龙凝胶膏剂对大鼠血清中NF-κB、TNF-α、IL-1β浓度的影响,筛选出调控作用最为显著的化合物并进行质量标准研究。方法:制备大鼠类风湿性关节炎(RA)模型,分别给予相同治疗量的桂龙凝胶膏剂和不同单体化合物溶液,取血,采用Western-Blot法和酶联免疫吸附法(ELISA)测定大鼠血清中NF-κB、TNF-α、IL-1β水平,根据不同组别上述指标的变化,确定桂龙凝胶膏剂中起调控作用的成分。质量标准研究采用HPLC法,以Agilent Eclipse XDB C18为色谱柱,以乙腈(A)-0.05%磷酸水溶液(B)为流动相,流速1.0mL/min,柱温25℃,检测波长:235nm。结果:血清指标测定结果表明,乌头碱、新乌头碱和桂皮醛对NF-κB、TNF-α、IL-1β水平影响最为显著;质量标准研究中,三种成分分离度符合要求,方法精密度、准确度、稳定性和回收率良好,10批桂龙凝胶膏剂样品含量测定结果显示,三种有效成分含量的RSD分别为4.60%、3.54%和8.80%。结论:本研究通过测定NF-κB、TNF-α、IL-1β水平,更有针对性地对桂龙凝胶膏剂中与功效相关的活性成分进行筛选和质量控制,为基于活性成分导向的质量标准和指纹图谱研究提供了参考。

Objectives： To establish activity oriented quality standard for Guilong gel, by observing its influence on NF-κB, TNF-α and IL-1β levels in rat serum; Mehtods： rat rheumatoid arthritis model was constructed and treating by using Guilong gel and various kinds of compounds, followed by NF-κB, TNF-α and IL-1β level tests using western blot and ELISA to determine compounds act as regulator; quality control research was carried out by HPLC, with a Agilent Eclipse XDB C18 column and acetonitrilephosphoric acid as the mobile phase; determination was carried out at the flow rate of 1.0mL/min, 25℃, with-the detection wavelength of 235nm; Results： As test of the serum indicated, aconitine, mesaconine and einnamaldehyde had most obvious influence on NF-κB, TNF-α and IL-1β levels; The three compound had satisfactory resolution, with good precision, accuracy, stability and recovery; content determination of 10 batches of the gel showed that the RSDs of the target compound was 4.60% ,3.54% and 8.80% respectively; Conclusions： through determination of NF-κB, TNF-α and IL-1β levels, this study focused on screening and quality control of effective related components of Guilong gel, providing helpful references for activity guided quality standard and fingerprint establishment.