北马兜铃根的化学成分研究

从北马兜铃根中分离到了５个化合物，经化学及光谱分析等方法鉴定为马兜铃酮，７－甲氧基马兜铃酸Ａ，７－羟基马兜铃酸Ａ，马兜铃酸Ｃ，马兜铃内酞胺－Ｎ－六碳糖甙。

Human AKR1A1 involves in metabolic activation of carcinogenic aristolochic acidⅠ

OBJECTIVE To investigate whether aldo-keto reductases(AKRs)can act as a nitrore⁃ductase(NR)and bioactivate aristolochic acidⅠ(AA-Ⅰ)to produce AA-Ⅰ-DNA adducts.METHODS①Human-induced hepatocytes(hiHeps)and human bladder RT4 cells were used as tool cells and treated with AA-Ⅰ0,0.5,1.0 and 2μmol·L^(-1)for 24 h.Cell viability was detected using the CCK-8 method,and the half maximal inhibition concentration(IC_(50))was calculated using the CCK-8 method and the level of DNA adduct production was calculated.②hiHeps and RT4 cells were treated with AKR inhibitor luteotin(0,5,10 and 25μmol·L^(-1))+AA-Ⅰ0.2 and 1.0μmol·L^(-1)for 24 h,respectively,and the levels of DNA adducts were detected by a liquid chromatography-tandem mass spectrometer(LC-MS/MS).③hiHeps cells were incubated with 80 nmol·L^(-1)small interfering RNAs(si-AKRs)for 48 h and treated with AA-Ⅰ1.0μmol·L^(-1)for 24 h.Real-time qualitative PCR(RT-qPCR)method was used to detect the mRNA expression of AKRs gene and LC-MS/MS technology was used to investigate the effect of specific AKR gene knockdown on DNA adduct levels.④500 nmol·L^(-1)human AKR recombinant proteins AKR1A1 and AA-Ⅰwere incubated in vitro under anaerobic conditions and the formation of AA-Ⅰ-DNA adducts was detected.RESULTS①The IC_(50)of AA-Ⅰto hiHeps and RT4 cells was 1.9 and 0.42μmol·L^(-1),respec⁃tively.The level of DNA adduct production of the two cell lines was significantly different(P<0.01).②Luteolin≥5μmol·L^(-1)significantly inhibited the production of AA-Ⅰ-DNA adducts in both cells(P<0.05),and there was a concentration-dependent effect in hiHeps cells(P<0.01,R=0.84).③In the AKR family,the knockdown of AKR1A1 gene up to 80%inhibited the generation of AA-Ⅰ-DNA adducts by 30%-40%.④The AA-Ⅰ-DNA adducts were detected in the incubation of recombinant protein AKR1A1 and AA-Ⅰunder anaerobic conditions in vitro,approximately 1 adduct per 107 nucleotides.CONCLU⁃SION AKR1A1 is involved in AA-Ⅰbioactivation,providing a reference for elucidation of the carcino⁃genic mechanism of AA-Ⅰ.

基于多成分含量测定结合化学计量法评价水族药材骂广瓦质量

目的:建立高效液相色谱法测定水族药材骂广瓦中甲基丁香酚、马兜铃酸Ⅱ、马兜铃内酰胺Ⅰ、马兜铃酸Ⅰ和马兜铃酮的含量。方法:采用Welch C18(4.6 mm×250 mm,5 μm),以乙腈-7.0 mmol/L磷酸缓冲溶液(pH=2.3)为流动相,梯度洗脱,流速:1.0 mL/min;检测波长:230 nm;柱温:40 ℃。采用SPSS 21.0及SIMCA 13.0软件进行综合分析评价骂广瓦药材的质量。结果:各组分在一定范围内呈线性关系,回收率在97.77%~100.16%之间,RSD在0.91%~2.19%之间;定量数据经统计学软件分析,甲基丁香酚、马兜铃酸Ⅱ、马兜铃内酰胺Ⅰ可作为影响差异的主要成分,OPLS-DA图也可直观的区别不同产地的样品。结论:该方法简便、准确可靠,可为骂广瓦的质量控制提供依据。