摘要
目的观察酒精对小鼠骨髓基质细胞中脂肪细胞特异性基因422(aP2)和Ⅰ型胶原基因表达的影响。方法采用原代骨髓基质细胞体外培养技术,取小鼠双侧股骨骨髓细胞,通过细胞贴壁、分离,获取骨髓基质细胞。以0.09mol/L的酒精浓度作为诱导剂处理细胞。采用完整细胞斑点印迹分子杂交方法检测实验组和对照组细胞中422(aP2)mRNA和Ⅰ型胶原mRNA的表达。结果对实验组和对照组的422(aP2)mRNA的完整细胞斑点印迹扫描值进行比较,实验组中422(aP2)基因杂交信号明显增强,斑点印迹扫描值为7207.8±331.3,对照组斑点印迹扫描值为652.2±62.6,实验组为对照组的11倍,两组比较差异有非常显著性意义(P<0.001)。对实验组和对照组Ⅰ型胶原mRNA的完整细胞斑点印迹扫描值进行比较,实验组基因杂交信号较对照组明显减弱,斑点印迹扫描值为3 567.3±300.9,对照组斑点印迹扫描值为7487.0±488.4,为实验组的2倍,两组比较差异有非常显著性意义(P<0.001)。结论酒精能诱导骨髓基质细胞向脂肪细胞分化,而减少骨髓基质细胞向成骨细胞分化。这可能与酒精性骨坏死的发生机制有关。
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 (P<0.001). 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 (P<0.001). 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-indu
出处
《中华骨科杂志》
CAS
CSCD
北大核心
2003年第8期493-495,共3页
Chinese Journal of Orthopaedics
