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 bla...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.展开更多
为研究有机酸中辅助官能团对磷石膏晶体调晶效果的影响,利用计算机仿真技术模拟琥珀酸、苹果酸和马来酸与CaSO_4·0.5H_2O(111)晶面的吸附反应,并利用吸附模型CASTEP(cambridge serial total energy package)计算上述3种有机酸与CaS...为研究有机酸中辅助官能团对磷石膏晶体调晶效果的影响,利用计算机仿真技术模拟琥珀酸、苹果酸和马来酸与CaSO_4·0.5H_2O(111)晶面的吸附反应,并利用吸附模型CASTEP(cambridge serial total energy package)计算上述3种有机酸与CaSO_4·0.5H_2O晶面相互作用的吸附能,得出它们对晶面调晶效果的优先顺序.通过CaSO_4·0.5H_2O晶面吸附有机酸前后有机酸中羧基键长、键角以及羧基与晶面的态密度变化,对比发现在琥珀酸羧基邻位引入富电子羟基基团或者在中间位引入顺式结构双键均有助于提高羧基与晶体表面Ca2^(+)的络合电子配位能力.利用扫描电镜(SEM)观察有机酸与磷石膏反应后的晶体形貌,并测得与有机酸反应后磷石膏试块的力学参数.结果表明:试验结果和计算机仿真技术模拟的结论一致;与苹果酸反应后的磷石膏试块抗压强度和抗折强度均最大,分别为12.85,4.21MPa.展开更多
文摘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.
文摘为研究有机酸中辅助官能团对磷石膏晶体调晶效果的影响,利用计算机仿真技术模拟琥珀酸、苹果酸和马来酸与CaSO_4·0.5H_2O(111)晶面的吸附反应,并利用吸附模型CASTEP(cambridge serial total energy package)计算上述3种有机酸与CaSO_4·0.5H_2O晶面相互作用的吸附能,得出它们对晶面调晶效果的优先顺序.通过CaSO_4·0.5H_2O晶面吸附有机酸前后有机酸中羧基键长、键角以及羧基与晶面的态密度变化,对比发现在琥珀酸羧基邻位引入富电子羟基基团或者在中间位引入顺式结构双键均有助于提高羧基与晶体表面Ca2^(+)的络合电子配位能力.利用扫描电镜(SEM)观察有机酸与磷石膏反应后的晶体形貌,并测得与有机酸反应后磷石膏试块的力学参数.结果表明:试验结果和计算机仿真技术模拟的结论一致;与苹果酸反应后的磷石膏试块抗压强度和抗折强度均最大,分别为12.85,4.21MPa.
