Optimization of extraction process of flavonoids from Broussonetia papyrifera leaves and its antioxidant effect on mouse epidermal stem cells
Objective To optimize the extraction process of flavonoids from Broussonetia papyrifera leaves and explore the antioxidant effect of flavonoids on mouse epidermal stem cells.Methods The extraction process of flavonoids from Broussonetia papyrifera leaves was optimized by single factor experiment,including the liquid-solid ratio(15∶1,20∶1,25∶1,30∶1 and 35∶1),sodium hydroxide(NaOH)concentration(0.2％,0.4％,0.6％,0.8％and 1.0％),pH value(2.5,3.0,3.5,4.0 and 4.5)and extraction temperature(60,65,70,75 and 80 ℃).Based on the results of single factor experiment,the optimal extraction process was determined by orthogonal test with the mass fraction of flavonoids as the evaluation index.CD49f+/CD71-mouse epidermal stem cells were isolated and cultivated by immunomagnetic bead method,and the effects of flavonoids on the cell relative viability and the contents of reduced glutathione(GSH)and malondialdehyde(MDA)were detected.Results The optimal extraction conditions of flavonoids were liquid-solid ratio of 30∶1,0.6％NaOH,pH 4.5 and extraction temperature of 75 ℃.Under these conditions,the average mass fraction of flavonoids extracted was 1.47％.Compared with the negative control group,when the flavonoids final concentration was 25 and 50 μg/mL,the cell relative viability increased significantly(F=1.427 and 13.747 respectively,each P＜0.01);when the final concentration of flavonoids was 12.5,25 and 50 μg/mL,the content of GSH increased significantly(F=0.044,0.291 and 2.577 re-spectively,each P＜0.05)and the content of MDA decreased significantly(F=3.568,4.909 and 1.400 respectively,each P＜0.05).Conclusion The optimized extraction process of flavonoids from B.papyrifera leaves was stable and reliable,which is beneficial to the reuse of remaining stock solution after processing,and the extracted flavonoids can promote the pro-liferation of mouse epidermal stem cells and perform antioxidant activity.
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