Optimization of complex enzymatic extraction process of steroidal glycosides from Cynanchum auriculatum Royle ex Wight and its hepatoprotective activity

Abstract

Objective: Response surface methodology was used to optimize the extraction process of steroidal glycosides from Cynanchum auriculatum Royle ex Wight by complex enzymatic method, and its effects on alcohol dehydrogenase activity and human hepatoma cell HepG2 were analyzed. Methods: Four experimental factors; enzymolysis duration, temperature, compound enzyme dose and pH were used as responses to a single-factor experiment of the Response Surface Methodology on the steroidal glycoside yields, using perennial C. auriculatum as the target material. The alcohol-induced hepatocyte damage model of HepG2 was used to investigate the contents of ALT, LDH, and GSH and changes in cell shape. Results: The results show that the optimum extraction conditions of steroidal glycoside enzyme were obtained at an enzymolysis duration of 100 min, a temperature of 46 ℃, enzyme dose of 1.30% and pH of 5.0. Consequently, the actual extraction rate of （2.80±0.04）% was achieved using the optimized process conditions, which was close to the expected value. Compared with the model group, it significantly suppresses the growth of ALT and LDH while increasing the amount of intracellular GSH, and the cell damage was restored. Conclusion: The compound enzyme method can effectively extract the steroidal glycosides of Cynanchum auriculatum Royle ex Wight; steroidal glycosides of C. auriculatum have hepatoprotective effect.

Keywords

• cynanchum auriculatum royle ex wight
• steroidal glycosides
• complex enzymatic method
• hepatoprotective activity