枯草杆菌发酵对何首乌抗氧化成分及肝癌细胞抗增殖作用的影响

Effects of Polygonum multiflorum Thunb fermented by Bacillus subtilis on main antioxidant ingredient contents and antiproliferation effects on hepatoblastoma cells

【目的】探讨何首乌经枯草杆菌发酵后主要抗氧化成分及其对肝癌细胞抗增殖作用效果的变化,为进一步研究何首乌发酵炮制方法及开发何首乌产品提供科学参考。【方法】将生何首乌用枯草杆菌发酵处理后,测定生品和发酵品的游离型蒽醌、结合型蒽醌和总多酚含量等成分的变化,并对比何首乌生品和发酵品对肝癌细胞HepG2抗增殖作用的变化。【结果】何首乌生品和发酵品总游离型蒽醌含量分别为1.56和1.60 mg/g,总结合型蒽醌含量分别为1.24和0.79 mg/g,何首乌经枯草杆菌发酵后结合型蒽醌含量极显著下降(P<0.01,下同)。何首乌生品和发酵品水提取液的总多酚含量分别为44.88和74.01 mg/g,乙醇提取液的总多酚含量分别为61.84和93.86 mg/g,何首乌经枯草杆菌发酵后总多酚含量极显著增加。0.025 g/L何首乌生品对肝癌细胞HepG2作用24 h后的细胞抑制率为4.2%,0.25 g/L何首乌生品对肝癌细胞HepG2作用72 h后的细胞抑制率为-11.1%(负值表示促进细胞生长),即随着何首乌生品浓度增加和作用时间延长,其对肝癌细胞生长的作用由抑制转变为促进。经枯草杆菌发酵后,0.025 g/L何首乌发酵品对肝癌细胞HepG2作用24 h后的细胞抑制率为9.9%,0.25 g/L何首乌发酵品对肝癌细胞HepG2作用72 h后的细胞抑制率为20.9%,即随着何首乌发酵品浓度增加和作用时间延长,其对肝癌细胞HepG2有明显的抗增殖效果。【结论】枯草杆菌发酵可降低何首乌的毒性,增强其对肝癌细胞HepG2的抗增殖作用,表明使用枯草杆菌发酵何首乌的炮制方法是一种安全有效的炮制减毒方法,对开发相应何首乌炮制品有重要意义。

【Objective】To investigate the changes of main antioxidant ingredients and inhibition effects on hepatoblastoma of Polygonum multiflorum Thunb(PM)after fermentation by Bacillus subtilis(BPM),and provide a scientific reference for further research of the PM fermentation method and exploring the PM fermentation products.【Method】After fermented by B.subtilis,changes of free anthraquinone,conjugated anthraquinone components and total polyphenol content in PM and BPM were determined,and changes of inhibition effects on hepatoblastoma HepG2 cell of PM and BPM were also investigated.【Result】In PM and BPM,free anthraquinone content were 1.56 and 1.60 mg/g,conjugated anthraquinone in PM and BPM were 1.24 and 0.79 mg/g.After fermentation,conjugated anthraquinone in BPM were decreased extremely significantly(P<0.01,the same below).The total polyphenol of PM and BPM water extract were 44.88 and 74.01 mg/g respectively,ethanol extract of PM and BPM were 61.84 and 93.86 mg/g respectively,it indicated that after fermentation by B.subtilis,the total polyphenol content were increased extremely significantly.0.025 g/L PM extract showed 4.2%inhibition to hepatoblastoma HepG2 cells after 24 h,0.25 g/L PM extract showed-11.1%inhibition to hepatoblastoma HepG2 cells after 72 h(negative number meaned growth promotion),it indicated that with the increasing of concentration and time,the effect of PM on the hepatoblastoma HepG2 cells changed from inhibition to promotion.And after fermentation by B.subtilis,0.025 g/L BPM extract exhibited 9.9%inhibition to hepatoblastoma HepG2 cells after 24 h,0.25 g/L BPM extract exhibited 20.9%inhibition to hepatoblastoma HepG2 cells after 72 h,it showed that with the increasing of concentration and time,BPM had an obvious anti-proliferation effect on the hepatoblastoma HepG2 cells.【Conclusion】These results show that fermentation by B.subtilis can reduce toxicity of PM and increase the anti-proliferation effects on hepatoblastoma HepG2 cells,which indicates that fermentation using B.subtilis can be helpful for the utilization of the BPM preparation method as a new potential procedure to process PM and reduce toxicity.