肝肾细胞毒性导引下鱼腥草中内酰胺生物碱的分离和鉴定

Isolation and Identification of Lactams from Houttuynia cordata Thunb.under Guidance of Hepatorenal Cytotoxicity

目的:对鱼腥草HouttuyniacordataThunb.地上部分化学成分及其肝、肾细胞毒性进行研究。方法:采用CCK-8法测定鱼腥草不同极性段萃取物对6种肝、肾细胞(小鼠正常肝细胞AML-12、大鼠肝细胞BRL-3A、人正常肝细胞L-02、大鼠肾小管上皮细胞NRK-52E、非洲绿猴肾细胞Vero和人肾皮质近曲小管上皮细胞HK-2)的毒性。利用硅胶、中性氧化铝、MCI和制备高效液相色谱法(HPLC)分离纯化,结合质谱(MS)和核磁共振(NMR)鉴定化合物的结构。结果:鱼腥草二氯甲烷部位较其他部位具有明显细胞毒性;从中分离并鉴定了6个化合物,分别为3,5-二癸酰基-4-壬基-1,4-二氢吡啶(1)、马兜铃内酰胺AⅡ(2)、马兜铃内酰胺FⅠ(3)、顺式-N-(4-羟基苯乙烯基)苯酰胺(4)、马兜铃内酰胺BⅡ(5)、头花千金藤二酮B(6)。鱼腥草二氯甲烷部位对AML-12、BRL-3A、L-02和NRK-52E细胞具有明显的细胞毒性;化合物2、3和5对部分细胞株具有一定的细胞毒性,化合物6对6种测试细胞均具有一定细胞毒性作用。结论:鱼腥草中内酰胺生物碱具有一定肝、肾细胞毒性。

Objective:To study the chemical components from the aerial part of Houttuynia cordata Thunb.and its hepatorenal cytotoxicity.Methods:Different polar extracts were evaluated for their hepatorenal cytotoxicity in normal murine hepatocytes AML-12,rat hepatocytes BRL-3A,normal human hepatocytes L-02,rat renal tubular epithelial cells NRK-52E,African green monkey kidney cells Vero and human renal cortex proximal tubule epithelial cells HK-2 cell lines by cell counting kit-8(CCK-8)assay.The chemical constituents were separated and identified by column chromatography on silica gel,neutral aluminum oxide,middle chromatogram isolated gel(MCI)and preparative high-performance liquid chromatograph(HPLC),while their structures were determined by physicochemical properties and spectroscopic data.Results:The dichloromethane fraction exhibited higher cytotoxicity as compared with other fractions.Six compounds were isolated from the dichloromethane fraction of the aerial parts of H.cordata Thunb.,and identified as 3,5-didecanoyl-4-nonyl-1,4-dihydropyridine(1),aristololactam AⅡ(2),aristololactam AⅡ(3),cis-N-(4-hydroxystyryl)benzamide(4),aristololactam BⅡ(5),cepharadione B(6).The dichloromethane fraction exhibited high cytotoxicity against AML-12,BRL-3A,L-02 and NRK-52E cells.Compounds 2,3,and 5 showed certain cytotoxicity to some cells.Compound 6 showed certain cytotoxicity to all the six cell lines.Conclusion:The lactams from H.cordata Thunb.had certain hepatorenal cytotoxicity.