Cordyceps militaris is a well-known traditional Chinese medicine. Studies have demonstrated that the polysaccharides of C. militaris have various bioactivities. However, their mechanisms of action remain unclear. We previously purified a water-soluble polysaccharide CM1 from C. militaris and found that it has a cholesterol efflux improving capacity. This study further investigates the effect of CM1 in anti-atherosclerosis and its underlying mechanism in apolipoprotein E-deficient mice. Our data indicated that CM1 significantly decreased the total cholesterol and triglyceride in the plasma of mice, and decreased lipid deposition and formation of atherosclerotic plaque in a dose-dependent manner. Integrated bioinformatics analysis revealed that CM1 interacted with multiple signaling pathways, including those involved in lipid metabolism, inflammatory response, oxidoreductase activity and fluid shear stress, to exert its anti-atherosclerotic effect. Molecular technology analysis showed that CM1 enhanced the expression of proteins involved in lipid metabolism, reduced the expression of intercellular adhesion molecule-1 and tumor necrosis factor-? in the aorta, and decreased the content of oxidative products by enhancing the activities of antioxidant enzymes. Microarray analysis and biochemical data indicated that CM1 can improve lipid metabolism, reduce inflammation and oxidative stress. Taken together, CM1 could be used for the treatment of hyperlipidemia and atherosclerotic cardiovascular diseases.

polysaccharide, microarray, atherosclerosis, bioinformatics analysis, cardiovascular disease

There is only one chemically distinct structure:

SVGMWSMILES3D molIonize

 

Warning = sub-repeat ending at bDGalf: unknown unit count was assumed to be 3
Warning = sub-repeat ending at aDManp: unknown unit count was assumed to be 3
[*]O[C@@H]1[C@@H](CO)O[C@@H](O[C@H]2[C@@H](O)[C@H](O)[C@@H](CO[C@H]3O[C@H](CO)[C@@H](O)[C@H](O)[C@@H]3O[C@H]3O[C@H](CO)[C@@H](O)[C@H](O)[C@@H]3O[C@H]3O[C@H](CO)[C@@H](O)[C@H](O)[C@@H]3O[C@H]3O[C@H](CO)[C@@H](O)[C@H](O)[C@@H]3O)O[C@@H]2O[C@H]2[C@@H](O)[C@H](O)[C@@H](COC)O[C@@H]2O[C@H]2[C@@H](O)[C@H](O)[C@@H](CO[C@@H]3O[C@@H]([C@H](O)CO)[C@H](O)[C@H]3O[C@@H]3O[C@@H]([C@H](O)CO)[C@H](O)[C@H]3O[C@@H]3O[C@@H]([C@H](O)CO)[C@H](O)[C@H]3O[C@@H]3O[C@@H]([C@H](O)CO)[C@H](O)[C@H]3O)O[C@@H]2O[C@@H]2[C@@H](CO)O[C@@H](O[C@@H]3[C@@H](CO)O[C@@H]([*])[C@H](O)[C@H]3O)[C@H](O)[C@H]2O)[C@H](O)[C@H]1O

2284.001 g/mol (C₈₅H₁₄₂R₂O₇₀, R = next and previous repeats, not counted in mol weight)

A new polysaccharide (CMPB90-1) was isolated from cultured Cordyceps militaris by alkaline extraction. The chemical structure of CMPB90-1 was determined by analysis of physicochemical and spectral data. The backbone of CMPB90-1 is composed of (1→6)-linked α-D-glucopyranosyl and (1→3)-linked α-D-glucopyranosyl residues, with branching at O-6, which consists of (1→4)-linked β-D-mannopyranosyl and (1→6)-linked α-D-glucopyranosyl residues, respectively. β-D-Galactopyranosyl residues is the terminal unit. In vitro immunomodulatory assay revealed that CMPB90-1 promoted proliferation of splenic lymphocytes, enhanced cytotoxicity of NK cells and promoted lymphocyte secretion of the cytokine interleukin-2. Besides, CMPB90-1 upregulated T-cell subpopulation, strengthened phagocytosis function of macrophages and induced their M1 polarization. The mechanism of the effects might be due to the activation of TLR2, MAPK and NF-κB pathways. The results proposed that CMPB90-1 can be researched and developed as a new functional food.

NMR, Structure determination, Immunostimulatory activity, Cordyceps

13C NMR data:
Linkage	Residue	C1	C2	C3	C4	C5	C6
3,6,6,4	bDGalp	105.01	72.62	76.02	71.34	75.82	62.43
3,6,6	bDManp	102.84	69.56	70.65	85.09	76.27	66.25
3,6	aDGlcp	101.83	76.18	85.24	70.53	76.26	66.35
3	aDGlcp	98.31	73.05	75.17	69.52	74.87	69.91
6,6,6,6	bDGalp	105.01	72.62	76.02	71.34	75.82	62.43
6,6,6	aDGlcp	98.31	73.05	75.17	69.52	74.87	69.91
6,6	aDGlcp	98.31	73.05	75.17	69.52	74.87	69.91
6	aDGlcp	98.31	73.05	75.17	69.52	74.87	69.91
	aDGlcp	101.83	76.18	85.24	70.53	76.26	66.35


1H NMR data:
Linkage	Residue	H1	H2	H3	H4	H5	H6
3,6,6,4	bDGalp	4.85	3.64	3.95	3.54	3.31	3.48
3,6,6	bDManp	4.53	3.63	3.82	3.67	3.83	3.81
3,6	aDGlcp	5.08	3.75	3.73	3.75	3.57	3.88
3	aDGlcp	5.00	3.87	3.45	3.34	3.63	3.98
6,6,6,6	bDGalp	4.85	3.64	3.95	3.54	3.31	3.48
6,6,6	aDGlcp	5.00	3.87	3.45	3.34	3.63	3.98
6,6	aDGlcp	5.00	3.87	3.45	3.34	3.63	3.98
6	aDGlcp	5.00	3.87	3.45	3.34	3.63	3.98
	aDGlcp	5.08	3.75	3.73	3.75	3.57	3.88


1H/13C HSQC data:
Linkage	Residue	C1/H1	C2/H2	C3/H3	C4/H4	C5/H5	C6/H6
3,6,6,4	bDGalp	105.01/4.85	72.62/3.64	76.02/3.95	71.34/3.54	75.82/3.31	62.43/3.48
3,6,6	bDManp	102.84/4.53	69.56/3.63	70.65/3.82	85.09/3.67	76.27/3.83	66.25/3.81
3,6	aDGlcp	101.83/5.08	76.18/3.75	85.24/3.73	70.53/3.75	76.26/3.57	66.35/3.88
3	aDGlcp	98.31/5.00	73.05/3.87	75.17/3.45	69.52/3.34	74.87/3.63	69.91/3.98
6,6,6,6	bDGalp	105.01/4.85	72.62/3.64	76.02/3.95	71.34/3.54	75.82/3.31	62.43/3.48
6,6,6	aDGlcp	98.31/5.00	73.05/3.87	75.17/3.45	69.52/3.34	74.87/3.63	69.91/3.98
6,6	aDGlcp	98.31/5.00	73.05/3.87	75.17/3.45	69.52/3.34	74.87/3.63	69.91/3.98
6	aDGlcp	98.31/5.00	73.05/3.87	75.17/3.45	69.52/3.34	74.87/3.63	69.91/3.98
	aDGlcp	101.83/5.08	76.18/3.75	85.24/3.73	70.53/3.75	76.26/3.57	66.35/3.88

There is only one chemically distinct structure:

SVGMWSMILES3D molIonize

 

[*]O[C@H]1[C@H](O)[C@@H](CO[C@@H]2O[C@H](CO)[C@@H](O[C@@H]3O[C@H](CO)[C@H](O)[C@H](O)[C@H]3O)[C@H](O)[C@@H]2O)O[C@H](OC[C@H]2O[C@H](O[C@@H]3[C@@H](O)[C@@H]([*])O[C@H](CO[C@H]4O[C@H](CO[C@H]5O[C@H](CO[C@H]6O[C@H](CO[C@@H]7O[C@H](CO)[C@H](O)[C@H](O)[C@H]7O)[C@@H](O)[C@H](O)[C@H]6O)[C@@H](O)[C@H](O)[C@H]5O)[C@@H](O)[C@H](O)[C@H]4O)[C@H]3O)[C@H](O)[C@@H](O)[C@@H]2O)[C@@H]1O

1459.269 g/mol (C₅₄H₉₀R₂O₄₅, R = next and previous repeats, not counted in mol weight)

Tumour-associated macrophages (TAMs) inhibit the killing effect of T lymphocytes on tumour cells through the immunocheckpoint programmed death ligand-1 (PD-L1)/programmed death-1 (PD-1) axis. TAMs-targeted therapy is a promising approach that could be used to reverse the immunosuppressive tumour microenvironment. Here, we further report CMPB90-1, a novel natural polysaccharide from Cordyceps militaris, could function as an anti-tumour modulator that resets TAMs from a tumour-promoting M2 phenotype to a tumour-killing M1 phenotype. This process involves reversing the functional inhibition of T lymphocytes by inhibiting the PD-L1/PD-1 axis between TAMs and T lymphocytes. Mechanistically, the membrane receptor of CMPB90-1 binding to M2 macrophages was identified by tandem mass spectrometry. CMPB90-1 converts immunosuppressive TAMs via binding to toll-like receptor 2 (TLR2), which causes the release of Ca2+ and the activation of p38, Akt and NF-κB, or ERK. This process then leads to the polarization of TAMs from M2 phenotype to the M1 phenotype. In vivo experiment shows that CMPB90-1 is able to polarize TAMs into the M1 phenotype and has anti-tumour effects with improved safety. Additionally, the anti-tumour effects of CMPB90-1 in vivo depend on the phenotypic conversion of TAMs. The results demonstrated that CMPB90-1 could be developed as a potential immune-oncology treatment reagent.

TLR2, Cordyceps militaris polysaccharide, PD-L1/PD-1, tumour-associated macrophages

← originally published structure   Show legend

There is only one chemically distinct structure:

SVGMWSMILES3D molIonize

 

[*]O[C@H]1[C@H](O)[C@@H](CO[C@@H]2O[C@H](CO)[C@@H](O[C@@H]3O[C@H](CO)[C@H](O)[C@H](O)[C@H]3O)[C@H](O)[C@@H]2O)O[C@H](OC[C@H]2O[C@H](O[C@@H]3[C@@H](O)[C@@H]([*])O[C@H](CO[C@H]4O[C@H](CO[C@H]5O[C@H](CO[C@H]6O[C@H](CO[C@@H]7O[C@H](CO)[C@H](O)[C@H](O)[C@H]7O)[C@@H](O)[C@H](O)[C@H]6O)[C@@H](O)[C@H](O)[C@H]5O)[C@@H](O)[C@H](O)[C@H]4O)[C@H]3O)[C@H](O)[C@@H](O)[C@@H]2O)[C@@H]1O

1459.269 g/mol (C₅₄H₉₀R₂O₄₅, R = next and previous repeats, not counted in mol weight)