BACKGROUND:In vitro experiments suggest that LIM mineralization protein-1 (LMP-1) gene can increase the expression of LMP-1 protein in osteoporotic bone marrow mesenchymal stem cells. OBJECTIVE:To investigate the LMP-1 expression in osteoporotic bone marrow mesenchymal stem cells transfected with RV-LMP-1-GFP in vitro. METHODS:Twelve SD female rats were selected and subjected to bilateral ovariectomy for establishment of osteoporosis models. After 2 months of feeding, bilateral femurs, tibiae, and humeri of rats were taken to isolate and culture bone marrow mesenchymal stem cells. Passage 3 cells were taken and randomly divided into ovariectomized group and LMP-1 transfection group. Another six rats only underwent removal of the same amount of fat tissue around the ovary, and passage 3 cells which were harvested as those in the former two groups served as sham group. RT-PCR and western blot assay were performed to determine the expression of LMP-1 protein and mRNA. RESULTS AND CONCLUSION:Under an inverted fluorescence microscope, transfected bone marrow mesenchymal stem cells from osteoporotic rats showed green fluorescent expression. The three groups al could express LMP-1 at the protein and mRNA levels. The expression levels of LMP-1 protein and mRNA in the LMP-1 transfection group were significantly higher than those in the ovariectomized group and sham group (P<0.05), but there was no statistical difference between the latter two groups (P>0.05). The successful expression of the RV-LMP-1-GFP gene in osteoporotic SD rat bone marrow mesenchymal stem cells was realized at the protein and mRNA levels;moreover, the expression of LMP-1 was increased dramatical y. These findings indicate that LMP-1 gene is successful y transferred into osteoporotic SD rat bone marrow mesenchymal stem cells, and significantly elevates the expression of LMP-1 mRNA and protein.

Objective To investigate the effects of dexamethasone on expressions of epithelial neutrophil activating protein-78 (ENA-78) and transforming growth factor- beta 1 (TGF-β1) in neutrophil of asthma rats.Methods Thirty male SD rats were randomly divided into three groups on average, including asthma group, control group and dexamethasone treated group. In this experiment, the rat model of asthma were established by sentization and challenge with ovalbumin. Blood neutrophil were isolated and purified. The expression of ENA-78 was detected by flow cytometry. The expression of TGF-β1 was detected by immunohistochemical method in blood neutrophil and bronchial wall. Results Expression of ENA-78 in blood neutrophil in dexamethasone treated group(71.82 ±8. 87 mean fluorescence intensity)was lower than that in asthma group, but higher than that in control group(all P ＜0. 01). And expressions of TGF-β1 protein in dexamethasone treated group(0. 173 ± 0. 014,0. 202 ± 0. 019 optical density, respectively) was lower than that in asthma group(all P ＜0. 01) ,but higher than that in control group(all P ＜0. 01). There were significant positive correlation between ENA-78 expression at blood neutrophil and numbers of total inflammation cells in bronchoalveolar lavage fluid (n = 29, γ = 0. 762, P ＜ 0. 01). Conclusion The beneficial effect of glucocorticoid(dexamethasone) on airway inflammation in asthma rats could be at least in part due to their direct inhibitory effect on ENA-78 and TGF-β1 protein generation by neutrophil.

Objective To observe the expressions of grouth-related oncogen (GRO)α, epithelial neutrophil activating protein-78 (ENA-78) and neutrophil-activating peptide-2 (NAP-2) of rat asthma. And to investigate the role of neutrophil in the pathogenesis of asthma exacerbation. Methods In this experi-ment, the rat model of asthma were randomly divided into two groups on average, including asthma group and control group. Levels of ENA-78 at blood neutrophil were detected by flow cytometry method. The ex-pressions of GROα protein at bronchial wall and NAP-2 protein at blood neutrophil were detected by immuno-histochemieal method. Results Levels of GROα, ENA-78 and NAP-2 proteins in asthma group [0.138 ±0.009(A value), 97.65±13.99(MFI), 0.198±0.016(A value), respectively]were significantly higher than those in control group[0.077±0.010(A value), 50.79±8.66(MFI), 0.079±0.015(A value), re-spectively], all P < 0.01. Conclusion Levels of GROα, ENA-78 and NAP-2 were increased at rat asth-ma. They may be participate in inflammation of asthma exacerbation. Neutrophil may promote inflammatory cells influxing into airway wall via increasing synthesis of CXC chemotactic factors.