Alcohol-induced regulation of adipogenic and osteogenic genes expression of marrow stromal cells

Objective The mechanism of osteonecrosis induced by alcohol was not clear, lipoidosis in bone cell could be one of reasons to cause osteonecrosis. The study was to observe the effect of alcohol on the adipocyte-specific gene, 422 (aP2), and osteoblastic gene, type-I collagen of marrow stromal cells in vitro experimentally. Methods The primary marrow stromal cells of the femur of 6 to 8 weeks mouse were procured and cultured in DMEM culture fluids, and the samples were isolated after adherent growth culture in vitro. The cells were divided into two groups, one of which was experimental group treated with 0.09 mol/L alcohol added with the culture fluids once for two days; another one was control group. The mixed culture discontinued 10 days later, the cells were collected after the combined handling of 0.22% EDTA and 0.25% trypsinase, and the concentration of the cell suspension was adjusted to 1×109/L. A volume of 10 μL of the cells suspension was dropped on the nitric cellulose membrane. The expression level of 422(aP2) and type-I collagen mRNA in the cells were investigated by means of intact cell RNA dot blot hybridization. Results The expression scanning value of the dot blot hybridization of 422(aP2)mRNA was 7 207.8±331.3 in the experimental group, that were as 11 times as that in the control group( 652.2±62.6), the difference was statistical significant between the two groups (P0.001). However, the expression value of type-I collagen mRNA contents in the experimental group was 3 567.3±300.9, and the value of type-I collagen mRNA contents in the control group was 7 487.0±488.4 and significantly higher than that of the experimental group, the difference was significant (P0.001). Conclusion The gene analysis of the research suggest that alcohol could induce marrow stromal cells to differentiate into adipocytes and inhibit osteogenic differentiation of the cells by regulating gene expression, which may be a important reason to cause lipoidosis in bone marrow and deficiency of bone repair in alcohol-induced osteinecrosis.