Study On The Isolation,Purification,Structure Identification And Biological Activity Of Cardamine Hupingshanensis Polysaccharides

Objective: In this paper,Enshi super-selenium-rich C.hupingshanensis was used as the raw material,through the extraction,separation and purification of C.hupingshanensis polysaccharides,the purpose of which was to clearly understand the structure of C.hupingshanensis polysaccharides,for the later stage Its structure-activity relationship research lays the foundation.Methods: Enzyme and ultrasonic assisted the extraction of C.hupingshanensis polysaccharides,and optimized the conditions using response surface methodology;purified C.hupingshanensis polysaccharides by column chromatography;then analyzed the structure of the isolated polysaccharides using corresponding structure analysis techniques;Finally,in vitro activity research method is used to explore biological activity of polysaccharides.Result: C.hupingshanensis polysaccharides were extracted by single factor and response surface method.The model obtained was significant,and the lack of fit was not significant.It was proved that the model was appropriate and the optimal conditions were obtained.The optimal conditions were liquid-to-solid ratio 30:1,ultrasonic time 24.80 min,cellulase ratio 2%,extraction temperature 90 �C,and extraction time 120 min.The extraction experiment was performed according to the optimal conditions to obtain the extraction amount of polysaccharide 48.53 mg/g;The crude polysaccharides of C.hupingshanensis were separated and purified to obtain 4 different polysaccharides,named CHP-1,CHP-2,CHP-3,CHP-4,and the structure analysis of the 4 polysaccharides showed that the main components of the first three polysaccharides were glucose,and the main component of CHP-4 was galactose,The molecular weights were 238.3 kDa,21.75 kDa,80.64 kDa,and 7.423 kDa,all of which contain a large number of ?-,?-glycosidic bonds and a small amount of uronic acid.The effects of crude polysaccharides on ABTS free radicals,hydroxyl free radicals and total reducing power were measured and compared with the same concentration of Vc.it could be concluded that crude polysaccharide of C.hupingshanensis had in vitro antioxidant activity,but at the same concentration was far less than the antioxidant capacity of Vc.All of them were semi-crystalline except for CHP-4 which was a full crystal structure;through the determination of free radical scavenging ability,it was found that the crude polysaccharide of C.hupingshanensis had a certain antioxidant activity;through in vitro cytological studies,it could be found that CHP-3 had cell activity,proliferation and migration of human lung cancer A549 the ability and invasion ability had obvious inhibitory effects;In the study of the apoptosis mechanism of A549 cells,it was found that by up-regulating the expression of P53,BAX and Caspase-3 genes in A549 cells,and down-regulating the expression of Bcl2 genes,cell apoptosis was caused.Conclusion: In this experiment,a new process for the rapid extraction of multiple elements from C.hupingshanensis was established.At the same time,a DEAE-52 cellulose chromatography column and a Sephadex G-100 dextran gel chromatography column were established for the separation and purification of C.hupingshanensis;The structure of the four polysaccharides was briefly discussed;and the in vitro anti-tumor activity of CHP-3 was confirmed.