Fig 1.
Ilomastat pretreatment influences the cytokines level.
The levels of TNF-a (A) and TGF-b1 (B) at day 10, 20 and 30, and SCF (C) at day 30 in mouse serum were measured by colorimetric enzyme-linked immunosorbent assay (ELISA). The final concentration of TGF-β1, TNF-α and SCF in each serum sample were corrected for sample dilution and presented in pg/mL. All error bars indicate SEM (n = 8). * P < 0.05; ** P < 0.01; *** P < 0.001.
Fig 2.
Ilomastat pretreatment mitigates radiation-induced BM failure in mice.
The bone marrows were collected from femoral bones of experimental mice and analyzed. (A) Panels show H&E staining of mouse femurs at day 10, 20 and 30 after 6 Gy γ-ray irradiation. Black arrowheads point at megakaryocytes. Representative images are shown from control, IR and IR + Ilomastat pretreatments. (B) Total nucleated cell numbers of BM and were scored in control, IR and IR + Ilomastat mice at day 10, 20 and 30 post TBI under light microscope. (C) Number of megakaryocytes in bone sections, determined by counting of 9 random fields per bone section for each mouse. All error bars indicate SEM (n = 8). * P < 0.05; ** P < 0.01; *** P < 0.001.
Fig 3.
Ilomastat pretreatment stimulates the cell proliferation of BM in irradiated mice.
Effect of Ilomastat on radiation mediated compromised proliferative ability in BM was evaluated by analysis of S-phase cells in the BM. Representative DNA flow cytograms (A). Frequency of S-phase cells in the BM of mice at day 3 (B) and 20 (C) post 6 Gy γ-ray irradiation. More than 10,000 BM cells were analyzed for each experiment and the experiments were repeated for three times independently. *P < 0.05, **P < 0.01. n = 8 in all groups.
Fig 4.
Ilomastat attenuates radiation-induced reduction of HSCs and HPCs.
Mice were administrated with/without 10 mg/kg Ilomastat 2 h before exposure to 6 Gy γ-ray irradiation. (A) A representative gating strategy of HSC and HPC analysis by flow cytometry. The frequencies of HSCs (B) and HPCs (C) in BMMCs were presented as mean ± SEM (n = 8). More than 10,000 BMMCs were analyzed for each experiment and the experiments were repeated for three times independently. **P < 0.05.
Fig 5.
Ilomastat pretreatment improves the spleen’s histological recovery of irradiated mice.
Mice were administrated with/without 10 mg/kg Ilomastat 2 h before exposure to 6 Gy γ-ray irradiation. H&E staining in the spleen of irradiated or unirradiated mice at day 10, 20 and 30 and the representative images were presented. Bar = 100 μm.
Fig 6.
Ilomastat improves the spleen weight and stimulates the splenic progenitor cells.
Mice were administrated with/without 10 mg/kg Ilomastat 2 h before exposure to 6 Gy γ-ray irradiation. (A) The morphology of spleen extracted from mice in each experimental group. (B) The changes of spleen weight in mice after irradiation. (C) The body weight of mice in each experimental group at indicated days after irradiation. (D) The spleen/body weight ratio changes in mice after irradiation. (E) Spleen was fixed in Bouin’s solution to enumerate the CFUs in IR and Ilomastat + IR. All error bars indicate SEM (n = 8). *P < 0.05; **P < 0.01; ***P < 0.001.
Fig 7.
The speculated model of Ilomastat promoting the recovery of radiation-induced hematopoietic injury in mice.
Ilomastat pretreatment decreases the TGF-β1 and TNF-α and increases the hematopoietic growth factor, directly or indirectly. Eventually, promote the recovery of radiation-induced hematopoietic injury.