[The effect of autologous bone marrow-derived cells mobilized by granulocyte colony stimulating factor on bleomycin-induced lung injury in mice.].
OBJECTIVE
To investigate the contribution of mobilized autologous bone marrow-derived cells (BMDC) to lung repair after lung injury induced by bleomycin, and the mechanisms of any protective effects conferred by BMDC.


METHODS
Sixty marrow-reconstructed mice were randomly divided into 2 groups: group A [bleomycin + granulocyte colony stimulating factor (G-CSF)] and group B (bleomycin + saline). Seventy-five normal mice were randomly divided into 3 groups: group C (bleomycin + G-CSF); group D (bleomycin + saline) and group N (saline only). Each group was further divided into 3 subgroups, which were sacrificed respectively on days 3, 7 and 14. Therapeutic evaluations were made by means of HE stain, Masson's trichrome stain, hydroxyproline concentration and pulmonary permeability index. The expressions of TGF-beta(1), IFN-gamma, MMP-9 and TIMP-1 in the lung tissue were detected by immunohistochemistry. Intrapulmonary BMDC was evaluated by flow cytometry and laser scanning confocal microscope. Another 20 mice were randomly divided into 2 groups including group E (bleomycin + G-CSF) and group F (bleomycin + saline). The survival time of each mouse was observed without end point.


RESULTS
The alveolitis score (mean rank 15.3), the pulmonary fibrosis score (46 +/- 8), the hydroxyproline concentrations (0.44 +/- 0.09) microg/mg, the TGF-beta(1) level (111 +/- 23), the IFN-gamma level (250 +/- 72) and the MMP-9 level (59 +/- 19) were significantly decreased in reconstructed treatment group on day 7 as compared to reconstructed control group, which was respectively (mean rank 28.0), (73 +/- 10), (0.52 +/- 0.07) microg/mg, (161 +/- 35), (299 +/- 31) and (314 +/- 77). Likewise, the alveolitis (mean rank 22.7), the pulmonary fibrosis (27 +/- 15), the hydroxyproline concentrations (0.41 +/- 0.05) microg/mg, the pulmonary permeability index (43.8 +/- 9.9) x 10(-3), the TGF-beta(1) level (132 +/- 55), the IFN-gamma level (178 +/- 23), and the MMP-9 level (101 +/- 54) in non-reconstructed treatment group on day 7 were significantly lower than those in non-reconstructed control group, (mean rank 33.9), (56 +/- 13), (0.49 +/- 0.08) microg/mg, (54 +/- 9) x 10(-3), (320 +/- 98), (409 +/- 61), (288 +/- 75), the differences being statistically significant (P < 0.05). The intrapulmonary BMDC level of reconstructed treatment group (0.65 +/- 0.13) was significantly higher than that in reconstructed control group (0.46 +/- 0.11), P < 0.05.


CONCLUSIONS
Mobilization of BMDC by G-CSF showed a protective effect on lung injury induced by bleomycin in mice, but did not have significant influence on survival time.
Jiang
Han-Shui
HS
Department of Respiratory Medicine, Affiliated Shanghai Pulmonary Hospital, Tongji University, Shanghai 200433, China.
Li
Hui-Ping
HP
Hu
Yang
Y
Chu
Jian-Xin
JX
Chen
Gang
G
Zhang
Rong-Xuan
RX
Zheng
Hui
H
D000818:Animals; D001761:Bleomycin; D001853:Bone Marrow; D016179:Granulocyte Colony-Stimulating Factor; D008168:Lung / Q000378:metabolism; D055370:Lung Injury; D051379:Mice; D011658:Pulmonary Fibrosis / Q000139:chemically induced
D001761:Bleomycin; D016179:Granulocyte Colony-Stimulating Factor