Sarcodon imbricatus polysaccharides improve mouse hematopoietic function after cyclophosphamide-induced damage via G-CSF mediated JAK2/STAT3 pathway

Abstract

Sarcodon imbricatus, a rare medicinal and edible fungus, has various pharmacological bioactivities. We investigated the effects of S. imbricatus polysaccharides (SIPS) on hematopoietic function and identified the underlying mechanisms using in vitro experiments with CHRF, K562, and bone marrow mononuclear cells (BMMNCs) and in vivo experiments with a mouse model of cyclophosphamide-induced hematopoietic dysfunction. We found that SIPS induced proliferation and differentiation of CHRF and K562 cells and upregulated the expression of hematopoietic-related proteins, including p90 ribosomal S6 kinases (RSK1p90), c-Myc, and ETS transcription factor, in the two cell lines. After 28 days of treatment, SIPS enhanced the bodyweight and thymus indices of the mice, alleviated enlargement of the spleen and liver, and contributed to the recovery of peripheral blood to normal levels. More importantly, the percentages of B lymphocytes and hematopoietic stem cells or hematopoietic progenitor cells were significantly elevated in bone marrow. Based on an antibody chip analysis and enzyme-linked immunosorbent assay, SIPS were found to successfully regulate 12 cytokines to healthy levels in serum and spleen. The cytokines included the following: interleukins 1Ra, 2, 3, 4, 5, and 6, tumor necrosis factor α, interferon−γ, granulocyte colony-stimulating factor (G-CSF) and macrophage colony-stimulating factor (M-CSF), C-C motif chemokine1, and monocyte chemoattractant protein−1. Moreover, SIPS upregulated the phosphorylation levels of janus kinase 2 (JAK2) and the signal transducer and activator of transcription 3 (STAT3) in the spleen, and similar results were validated in CHRF cells, K562 cells, and BMMNCs. The data indicate that SIPS activated the JAK2/STAT3 pathway, possibly by interactions among multiple cytokines, particularly G-CSF. We found that SIPS was remarkably beneficial to the bone marrow hematopoietic system, and we anticipate that it could improve myelosuppression induced by long-term radiotherapy or chemotherapy.

ZhenQi FuZheng Formula Improves Mouse Hematopoietic Function after Cyclophosphamide-Induced Damage via Enhancing Macrophage Colony-Stimulating Factor Concentrations

“…Similar as our previous study [20], the pathological alterations of spleen of mice with immunosuppression were detected using the hematoxylin-eosin (H&E) staining under the inverted microscope CKX41 (Olympus, Tokyo, Japan).…”

“…The seeded CHRF or K562 cells were incubated with 0, 25, or 100 µg/mL ZQFZ for 24 h, and the Cellular proliferation was detected by 2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H -tetrazolium-5carboxanilide (XTT) method as described previously [20].…”

“…Seeded K562 cells (4 × 10 5 cells/well) were incubated 0, 25, and 100 µg/mL of ZQFZ for 24 h. The erythroid differentiation and hemoglobin (HGB) of K562 cells were detected using Benzidine staining the same as our previous study [20].…”

“…The experimental protocol (2017SY0603) has been approved by the Institution Animal Ethics Committee of Jilin University. Seventy 7-week male BALB/c mice weighted 18-22 g (speci c pathogen-free (SPF) grade, SCXK(Liao)-2015-0001) were housed in a controlled environment the same as our previous study [20].…”

“…Under the sterile condition, the tibial and femoral section were collected immediately. Similar to our previous research, the ACK lysis buffer was used for the isolation of the bone marrow mononuclear cells (BMMNCs) [20]. The antibodies related to surface markers including FITC-conjugated anti-mouse Lineage Cocktail (133302), APCconjugated anti-mouse CD19 (152410), and PerCP-conjugated anti-mouse CD45 (103129) were applied to stain with cells at 25 °C for 15 min darkly.…”