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Mar Drugs
2017 Nov 01;1511:. doi: 10.3390/md15110341.
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Cucumarioside A₂-2 Causes Macrophage Activation in Mouse Spleen.
Pislyagin EA
,
Manzhulo IV
,
Gorpenchenko TY
,
Dmitrenok PS
,
Avilov SA
,
Silchenko AS
,
Wang YM
,
Aminin DL
.
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The immunomodulatory effect of triterpene glycoside cucumarioside A₂-2 (CA₂-2), isolated from the Far Eastern sea cucumber Cucumaria japonica, was compared with lipopolysaccharide (LPS) on mouse spleen. It has been shown that the intraperitoneal (i.p.) glycoside administration leads to increased spleen macrophage activating markers iba-1, IL-1β, iNOs, ROS and NO formation, with additional change of macrophage phenotype to M1. The mass spectrometry profiles of peptide/protein were obtained using MALDI-TOF-MS on the different parts of spleen sections isolated by laser mircodissection techniques. It was found that i.p. stimulation of animals with CA₂-2 leads to marked changes in the intensity of the characteristic peaks of spleen peptides/proteins, primarily in red pulp.
Figure 4. Macrophage M1/M2 polarization and activation. Flow cytometry analyses of RAW 264.7 cells treated with CA2-2 (0.1 or 0.01 μM), following by LPS (1 μg/mL) and IFN-γ (20 ng/mL) for M1 polarization or IL-4 (20 ng/mL) for M2 polarization.
Figure 5. The process of cutting red and white pulp in the mouse spleen section by laser microdissection (Carl Zeiss PALM MicroBeam). Green circles indicate the white pulp excision area; blue circles indicate the red pulp excision area. It can be seen that some of the selected tissue sections have already been cut (A); The bottom of the tube cap with cut out and collected tissue sections (B); Representative MALDI-TOF mass spectra of the mouse spleen samples from white pulp (C) and red pulp (D); The protein profiles of mouse treated with CA2-2 are shown. The pseudo-gel view of representative mass spectra of two different mouse spleen samples (Control and LPS) with all individual spectra shown in a density scale, in the mass range from 5000 to 6000 Da (E) and corresponding mass spectra profile, showing significant differences between untreated and LPS treated animal samples (F).
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