To establish an HPLC method for separation and determination of stigmasterol,β-sitosterol and lupeol in phyllanthus urinaria L by HPLC.A Eclipse Zorbax XDB-C18 column(4.6 mm×150 mm i.d.,5 μm) was used for chrom...To establish an HPLC method for separation and determination of stigmasterol,β-sitosterol and lupeol in phyllanthus urinaria L by HPLC.A Eclipse Zorbax XDB-C18 column(4.6 mm×150 mm i.d.,5 μm) was used for chromatographic separation under the conditions with methanol as the mobile phase at flow rate of 0.7 mL/min.The detection wavelength was 210 nm.The results indicated that stigmasterol,β-sitosterol and lupeol could be baseline separated.The linear range of stigmasterol,β-sitosterol and lupeol were 0.4mg/mL～8.8 mg/mL(R2 were 0.9975),0.5～10.0 mg/mL(R2 = 0.9978) and 0.5mg/mL～10.0 mg/mL(R2 = 0.9981),respectively.The mean recoveries of stigmasterol,β-sitosterol and lupeol were 94.4%～95.1%,91.2%～97.9% and 92.4%～104.6 %,respectively.RSD was less than 5%.The method has been applied to monitor the extraction of stigmasterol,β-sitosterol and lupeol in phyllanthus urinaria L by super-critical fluid extraction with satisfactory.展开更多
应用基于气相色谱-质谱联用(GC-MS)的代谢组学方法结合细胞周期实验,研究羽扇豆醇体外抑制人乳腺癌细胞MCF-7增殖的作用机理。代谢组学的研究结果表明:通过正交偏最小方差判别分析(OPLS-DA)可以很好地区分羽扇豆醇作用的MCF-7细胞代谢...应用基于气相色谱-质谱联用(GC-MS)的代谢组学方法结合细胞周期实验,研究羽扇豆醇体外抑制人乳腺癌细胞MCF-7增殖的作用机理。代谢组学的研究结果表明:通过正交偏最小方差判别分析(OPLS-DA)可以很好地区分羽扇豆醇作用的MCF-7细胞代谢谱与对照组细胞代谢谱,模型参数为:R2 Ycum=0.988,Q2 Ycum=0.964。VIP(variable importance in the projection)值大于1的差异代谢物进一步用t检验进行单位分析,选择t<0.05(VIP>1)的代谢物作为羽扇豆醇作用组的生物标志物,得到琥珀酸、磷酸、亮氨酸、异亮氨酸等11种代谢差异物。结合羽扇豆醇将细胞周期抑制在G1期这一现象,推测羽扇豆醇可能是主要抑制了三羧酸循环中的琥珀酰辅酶A的生成和底物磷酸化生成ATP的反应来抑制MCF-7细胞的增殖。本实验从代谢组学角度为乳腺癌抗肿瘤机制提供新的线索。展开更多
文摘目的基于网络药理学和分子对接探讨羽扇豆醇(Lupeol)治疗类风湿关节炎(Rheumatoid arthritis,RA)的作用机制。方法通过Swiss和TCMSP分析平台获取羽扇豆醇的作用靶点,在GeneCards数据库检索“RA”获取其相关靶点,并转化为相应的标准化基因名。利用R包绘制羽扇豆醇和RA共同靶点的韦恩图,获取交集基因。将交集基因导入STRING数据库,构建蛋白-蛋白相互作用(Protein-protein interactions,PPI)网络。通过clusterProfiler数据库进行京都基因(Geneontology,GO)和基因组百科全书(Kyoto encyclopedia of genes and gnomes,KEGG)通路富集分析。通过分子对接评价羽扇豆醇与AR、CASP3和CCNB1的结合作用。构建RA小鼠模型,测量各组小鼠的足体积,HE染色检测病理学变化,ELISA试剂盒检测小鼠血清TNF-α、IL-1β和IL-6的表达水平,采用Western blot检测各组小鼠Bcl-2、Bax、Casp3、Casp9的蛋白表达水平。结果获取羽扇豆醇40个作用靶点,RA疾病4734个相关靶点,羽扇豆醇-RA共同靶点27个,PPI网络自由度靠前的3个基因为AR、CASP3和CCNB1,GO富集结果是291个和KEGG通路富集为20条信号通路。分子对接显示羽扇豆醇与AR、CASP3和CCNB1亲和作用较好。第8、12、16、20天模型组小鼠足体积显著高于正常对照组(P<0.05),相较于模型组,羽扇豆醇组在第8、12、16、20天时小鼠足体积显著降低(P<0.05),双氯芬酸钠组在第12、16、20天时小鼠足体积显著降低(P<0.05)。HE染色结果表明,与模型组相比,羽扇豆醇药物组明显改善病理学状态。与模型组相比,羽扇豆醇药物组小鼠血清中TNF-α、IL-1β和IL-6水平降低(P<0.01)。Western blot结果显示,与正常组相比,模型组小鼠Bax、Casp3和Casp9蛋白表达显著降低(P<0.05),Bcl-2的蛋白表达显著升高(P<0.05)。与模型组比较,双氯芬酸钠组和羽扇豆醇药物组小鼠Bax、Casp3和Casp9蛋白表达显著上调(P<0.05),Bcl-2蛋白表达显著下调(P<0.05)。结论羽扇豆醇可通过调控p53信号通路中的Bax、Bcl-2、Casp3和Casp9从而发挥对RA的治疗作用。
文摘To establish an HPLC method for separation and determination of stigmasterol,β-sitosterol and lupeol in phyllanthus urinaria L by HPLC.A Eclipse Zorbax XDB-C18 column(4.6 mm×150 mm i.d.,5 μm) was used for chromatographic separation under the conditions with methanol as the mobile phase at flow rate of 0.7 mL/min.The detection wavelength was 210 nm.The results indicated that stigmasterol,β-sitosterol and lupeol could be baseline separated.The linear range of stigmasterol,β-sitosterol and lupeol were 0.4mg/mL～8.8 mg/mL(R2 were 0.9975),0.5～10.0 mg/mL(R2 = 0.9978) and 0.5mg/mL～10.0 mg/mL(R2 = 0.9981),respectively.The mean recoveries of stigmasterol,β-sitosterol and lupeol were 94.4%～95.1%,91.2%～97.9% and 92.4%～104.6 %,respectively.RSD was less than 5%.The method has been applied to monitor the extraction of stigmasterol,β-sitosterol and lupeol in phyllanthus urinaria L by super-critical fluid extraction with satisfactory.
文摘应用基于气相色谱-质谱联用(GC-MS)的代谢组学方法结合细胞周期实验,研究羽扇豆醇体外抑制人乳腺癌细胞MCF-7增殖的作用机理。代谢组学的研究结果表明:通过正交偏最小方差判别分析(OPLS-DA)可以很好地区分羽扇豆醇作用的MCF-7细胞代谢谱与对照组细胞代谢谱,模型参数为:R2 Ycum=0.988,Q2 Ycum=0.964。VIP(variable importance in the projection)值大于1的差异代谢物进一步用t检验进行单位分析,选择t<0.05(VIP>1)的代谢物作为羽扇豆醇作用组的生物标志物,得到琥珀酸、磷酸、亮氨酸、异亮氨酸等11种代谢差异物。结合羽扇豆醇将细胞周期抑制在G1期这一现象,推测羽扇豆醇可能是主要抑制了三羧酸循环中的琥珀酰辅酶A的生成和底物磷酸化生成ATP的反应来抑制MCF-7细胞的增殖。本实验从代谢组学角度为乳腺癌抗肿瘤机制提供新的线索。
