基于Nrf2-ARE通路的抗辐射有效活性成分筛选研究

Screening antioxidants to protect radiation via Nrf2-ARE reporter gene

目的建立基于Nrf2-ARE通路的报告基因筛选模型,筛选抗辐射复方四物汤加味方中的活性成分。方法将抗氧化反应元件插入萤火虫荧光素酶载体pGL4.26,构建重组质粒pGL4-ARE,与海肾荧光素酶载体pRL-TK共同转染HEK-293细胞,检测四物汤加味方中刺五加皂苷E、金丝桃苷、五味子乙素等12个主要的化学成分对Nrf2-ARE通路的激活作用。结果在四物汤加味方中,白藜芦醇(50μmol.L-1以上)和金丝桃苷(100μmol.L-1以上)的抗氧化活性最强,与空白对照组比较差异有显著性(P<0.01);红景天苷也有较好的效果,500μmol.L-1以上可提高诱导表达倍数(P<0.01)。结论白藜芦醇、金丝桃苷、红景天苷是四物汤加味方中的主要抗氧化活性成分。

Aim To establish a screening model based on Nrf2-ARE in order to screen antioxidants in Si-wu-tang derived compound,a radiation protective Chinese medicine formula.Methods A recombinant reporter vector pGL4-ARE was constructed.The HEK-293 cells were transiently transfected with a plasmid barboring the ARE-promoter（pGL4-ARE） and a control vector（pRL-TK）.Twelve chemical constituents including hyperoside,salidroside,resveratrol in new Si-wu capsule were exerted to treat reporter vector transfective cells,and the chemiluminescent intensity which produced in chemical constituent treated cells was measured.Results Resveratrol upon 50 μmol·L-1 and hyperoside upon 100 μmol·L-1 could activate Nrf2-ARE reporter gene（P0.01）,and salidroside upon 500 μmol·L-1 could also activate Nrf2-ARE reporter gene（P0.01）.Conclusion Resveratrol,hyperoside and salidroside are antioxidant chemical constituents in Si-wu-tang derived compound.