Studies On Structure Identification,Biological Activity Of Polysaccharide (CIS-A) From Catathelasma Imperiale (Fr.) Sing.and Polysaccharide (LSM-X) From Lactikporus Supharells (Fr.) Murr

Catathelasma imperiale(Fr.)Sing belongs to Agaricales,Tricholomataceae,pine bud mushroom genus.Catathelasma imperiale(Fr.)Sing has large fruiting bodies,which is a dual characteristic of edible and medicinal.Lactikporus supharells(Fr.)Murr belongs Dicotyledoneae,homobasidiomycetidae,Aphyllophorales,Polyporaceae,sulfur bacteria.It is an edible fungus that regulates the body and improves health.The present study for Catathelasma imperiale(Fr.)Sing and Lactikporus supharells(Fr.)Murr mainly concentrated in the domestication and antibacterial activity of secondary metabolites.However,the purification,structure and immunomodulatory activity of the polysaccharide have not been reported.In this paper,subentities of Catathelasma imperiale(Fr.)Sing and Lactikporus supharells(Fr.)Murr were used as experimental materials,respectively.Hot water extraction method was used to extract and purify the polysaccharides from Catathelasma imperiale(Fr.)Sing and Lactikporus supharells(Fr.)Murr.The structures of two fungal polysaccharides were identified by HPGPC,FT-IR,HPLC,NMR and GC-MS methods.In addition,the biological activity of CIS-A was investigated by CCK-8,flow cytometry,neutral red,Griess,fluorescence quantitative,cell model in vitro and S180 tumor model in vivo.The biological activity of LSM-X was investigated by CCK-8 and flow cytometry.The results of the study are as follows:(1)The polysaccharide was extracted from fruiting bodies of Catathelasma imperiale(Fr.)Sing and was named CIS-A.It was a homogeneous polysaccharide with a weight average molecular weight of 50486 Da.CIS-A is composed of glucose and fucose,and the ratio is 3:2.It had a backbone of 1,3-linked α-D-glucose which branches were mainly composed of two 2-linked β-L-fucose residue,and the β-L-fucose was 2,3-linked.(2)The polysaccharide was extracted from fruiting bodies of Lactikporus supharells(Fr.)Murr and was named LSM-X.It was a homogeneous polysaccharide with a weight average molecular weight of 14695 Da.LSM-X is composed of glucose,galactose and lyxose,and the ratio is 4:2:1.The main chain was 1,6-linked β-D-glucose,the 4-O had a side chain of two 1,6-linked α-D-galactose residues and a 1-linked α-D-lyxose residue.(3)The immunomodulatory activity of CIS-A shows that compared with the blank control group,CIS-A could promote the proliferation of three immune cells in the concentration range of 0.625-20 μg/mL.When the CIS-A concentration of polysaccharide was 10 μg/mL,the OD value of B lymphocyte proliferation activity reached the maximum,the rate of proliferation was 73.39%.When the concentration of CIS-A polysaccharide is 20 μg/m L,the proliferation activity of T is the best,the rate of proliferation was 98.02%.And when the concentration of CIS-A polysaccharide is 5 μg/m L,the proliferation activity of RAW264.7 cells is the best,the rate of proliferation was 58.63%.CIS-A polysaccharides can promote the proliferation of B cells by increasing the percentage of B cells in the S phase.Compared with the control group,the concentration of CIS-A polysaccharide in the range of 2.5-20 μg/mL could significantly promote macrophage phagocytosis activity(P<0.01),and had a dose-dependent effect.When the concentration of CIS-A polysaccharide was 20 μg/mL,the phagocytosis effect of RAW264.7 cells was the best.When the concentration of CIS-A polysaccharide was 40 μg/m L,the release of NO was the most in RAW264.7 cells.And CIS-A polysaccharide stimulated RAW264.7 cells to increase the relative expression of related genes(IL-1 beta,IL-6,IL-8,INOS,TNF-alpha)mRNA.(4)The antitumor activity of CIS-A showed that CIS-A could inhibit the growth of L929 cells in varying degrees in the concentration range of 2.5-40 μg/m L,and at 5 μg/mL,the survival rate of L929 cells was the lowest,only 69%.CIS-A polysaccharides can inhibit the growth of S180 tumor in the body,the inhibition rate was 64.4%.and no toxic and side effects have been found.(5)The immunomodulatory activity of LSM-X showed that compared with the blank control group,when LSM-X polysaccharides were 10 μg/mL,the proliferation of B cells was promoted effectively,and the proliferation rate was 48.35%.The proliferation rate was more than 20.20% compared with the positive control group(5 μg/mL LPS).when LSM-X polysaccharides were 10 μg/mL,the proliferation of T cells was promoted effectively.And the proliferation rate was 22.98%.The effect of proliferation was superior to the positive control group(5 μg/m L LPS).The research results showed CIS-A polysaccharide could promote the proliferation of immune cells(B lymphocytes,T lymphocytes and RAW264.7 cells),could promote the proliferation of B cells by regulating B cell cycle,improve the phagocytosis and releasing the NO level of RAW264.7 cells,and inhibit the L929 cell line in vitro,and also have a good inhibition on S180 tumor in vivo;LSM-X polysaccharides also promote the proliferation of immune cells(B lymphocytes,T lymphocytes and RAW264.7 cells).This study provides methods and techniques for the extraction and purification of polysaccharide from Catathelasma imperiale(Fr.)Sing and Lactikporus supharells(Fr.)Murr,and also provides experimental evidence for the enhancement of immune activity and antitumor activity of these two polysaccharides,which provides a theoretical basis for further development of the Catathelasma imperiale(Fr.)Sing and Lactikporus supharells(Fr.)Murr.In particular,it has important practical value and application value to improve human immune function and anti-tumor effect.However,more detailed immunoregulation mechanisms and anti-tumor mechanisms need further exploration...