We examined the effect of epidermal growth factor (EGF) treatment in mice that received bone marrow transplantation (BMT) after 11 Gy whole-body irradiation. C57Bl/6 mice were divided into three treatment groups: 0 Gy; 11 Gy ((60)Co, single dose, 0.51 Gy/min) with BMT (5 × 10(6) bone marrow cells isolated from green fluorescent protein syngeneic mice, 3-4 h postirradiation); and 11 Gy with BMT and EGF (2 mg/kg applied subcutaneously 1, 3 and 5 days postirradiation). Survival data were collected. Bone marrow, peripheral blood count and cytokines, gastrointestine and liver parameters and migration of green fluorescent protein-positive cells were evaluated at 63 days postirradiation. Epidermal growth factor increased survival of irradiated animals that received BMT from 10.7 to 85.7% at 180 days postirradiation. In the BMT group, we found changes in differential bone marrow and blood count, plasma cytokine levels, gastrointestinal tissues and liver at 63 days postirradiation. These alterations were completely or in some parameters at least partially restored by epidermal growth factor. These findings indicate that epidermal growth factor, administered 1, 3 and 5 days postirradiation in combination with bone marrow transplantation, significantly improves long-term prognosis.
The aim of our study was to examine the in vivo expression of p21 and expression and activation of ATF-2, c-Myc, and CREB in rat peripheral blood mononuclear cells (PBMC) after whole body γ-irradiation and to assess its contribution to biodosimetry. For Western blot experiments, male Wistar rats were whole-body irradiated by a single dose of 0, 0.5, 1, 3, and 5 Gy (60 Co, 1 m, 0.7 Gy/min). As a positive control, leukaemic MOLT-4 cells were used. For ELISA experiments, male Wistar rats were whole-body irradiated by a single dose of 0, 1, 2, 3, 4, and 5 Gy (60 Co, 1 m, 0.6 Gy/min). Blood samples were taken 4 h after the irradiation and PBMC were isolated using centrifugation on Histopaque-1077. Expressions of p21, ATF-2, phospho-ATF-2 Thr69/71 , c-Myc, phospho-c-Myc Thr-58/Ser62 , CREB, and phospho-CREB Ser133 were measured using Western blot method. Expression of p21 was also quantified using ELISA. We observed increase of p21 expression in rat PBMC 4 h after irradiation. According to ELISA, p21 levels increased 2.0-, 3.1-, 5.5-, 3.0-, and 3.1fold after irradiation by 1, 2, 3, 4, and 5 Gy, respectively. We did not detect any expression or activation of ATF-2, c-Myc and CREB. Protein p21 could be considered as a perspective biodosimetric marker of clinically significant irradiation (≥1 Gy) in vivo in unstimulated PBMC.
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