| Cordyceps cicadae is one of the most valuable Traditional Chinese medicines with a application history of more than 1600 years.It is an insect fungus complex formed by fungi parasitizing on cicada larvae.It belongs to the Claviciptaceae family and the Cordyceps genus.C.cicadae contains a variety of active components,including cordycepin,N-(2-hydroxyethyl)adenosine,cordycepin,polysaccharide,ergosterol and so on.Polysaccharide is one of its main active ingredients,which can improve immunity,reduce blood sugar,anti-tumor,protect kidney,antioxidant,anti-aging and other functions.Currently,the artificial culture technology of C.cicadae has made some progress,but researches of the active components and functions of the artificial culture of C.cicadae is still not deep enough.In this study,silkworm pupae(Bombyx mori)and tussah pupae(Antherea pernyi)were used as hosts to culture C.cicadae,and the polysaccharides of silkworm chrysalis C.cicadae and wild C.cicadae were extracted by ultrasonic assisted extraction method.Three kinds of crude polysaccharides were purified by activated carbon decolorization,Sevage method,dialysis and DEAE-52 cellulose column chromatography.Tussah pupae C.cicadae polysaccharides(TCCP),silkworm pupae C.cicadae polysaccharides(SCCP)and wild C.cicadae polysaccharides(WCCP)were obtained.The results of physicochemical properties and functional properties of three kinds of polysaccharides showed that the three kinds of polysaccharides had different oil holding capacity.The oil holding capacity of SCCP and WCCP was significantly higher than that of TCCP(P < 0.01),but there was no significant difference between SCCP and WCCP(P > 0.05);the three kinds of polysaccharides all had good water solubility;the results of foaming experiment showed that the foaming ability of the three kinds of polysaccharides was poor,and the foaming stability was also poor,which was less than 3%,which may be related to the larger molecular weight of the polysaccharides;SCCP had the highest emulsifying ability and emulsifying stability among the three kinds of polysaccharides.Antioxidant experiments in vitro showed that the three kinds of polysaccharides had good antioxidant activity and basically the same.With the increase of the concentration of TCCP,SCCP and WCCP,the scavenging activity of DPPH,ABTS and hydroxyl radicals,the chelating capacity of ferrous ions and the total reducing power of TCCP,SCCP and WCCP were increased in a concentration dependent manner.Finally,the proliferation inhibition and cytotoxicity of TCCP and WCCP on Hela cells were compared.The results showed that the polysaccharide from silkworm chrysalis C.cicadae and wild C.cicadae had direct cytotoxicity on tumor cells.Flow cytometry results showed that TCCP could block the S phase of Hela cells,and when TCCP was 1200 μg/ml,it could significantly induce apoptosis(P < 0.01),the total apoptosis rate was 73.17 %.The m RNA and protein expression of genes related to cell cycle and apoptosis signaling pathway were studied by RT-q PCR and Western blot.The results showed that TCCP could inhibit the expression of Cyclin E1,Cyclin A2 and CDK2,and up-regulated the expression of P53.In addition,Caspase family cascade is activated,the expression of death receptor is up-regulated,and the ratio of pro-apoptotic factor to anti-apoptotic factor is up-regulated,which leads to cell cycle arrest and apoptosis.The above results provided a theoretical basis for the development of artificial culture C.cicadae and the development of functional food or medicine,and increased the value of silkworm chrysalis provide the possibility for the economic development of silkworm industry. |
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