阿仑膦酸钠对小鼠骨髓基质细胞分化脂肪细胞瘦素表达的影响

INFLUENCE OF ALENDRONATE ON THE EXPRESSION OF LEPTIN OF ADIPOCYT ES DERIVED FROM MURINE MARROW STROMAL CELLS

目的 探讨阿仑膦酸钠 (ALN)对髓内脂肪细胞瘦素表达的影响。方法 采用原代小鼠骨髓基质细胞 (MSCs)培养诱导培养脂肪细胞技术 ,模拟髓内脂肪细胞的分化过程 ,在相同成脂诱导条件下 ,分别以 0mol/L(对照组 )、1× 10 -7mol/L(低剂量组 )、1× 10 -5mol/L(高剂量组 )ALN干预诱导成脂过程 ,Western blot法检测各组脂肪细胞瘦素的表达情况。结果 在诱导培养 4d后 ,Western blot结果显示各组蛋白条带光密度值分别为 0 80 98± 0 10 4 2、0 5 5 31± 0 0 5 34、0 4 12 5± 0 0 35 4。随着ALN浓度的增加 ,瘦素表达量依次降低 ,经方差分析显示各组差异具有统计学意义 (P <0 0 1)。结论 ALN可通过抑制髓内脂肪细胞瘦素表达而影响髓内脂肪细胞的功能。

Objective To explore the effect of alendronate on leptin expression of adipocytes that were derived from murine marrow stromal cells. Method To simulate the differentiation process of murine marrow ad ipocytes, primary marrow stromal cells (MSCs) were procured from the femur of 6-week mice and cultured in F12 HAM'S culture medium containing 10% fetal calf serum until the MSCs attained 80% confluence, and then the MSCs were cultured in F12 HAM'S medium containing 10% new-born culf serum and adipogenic agents for four days (10ng/L insulin, 10μM Dexamethasone, 05μM IBMX in the first 2 da ys and only 10ng/L insulin in the later 2 days). The adipocytes were identified by morphology and Oil Red O stain. During the adipogenic process, the cells were divided into three groups, two of which were experimental groups treated with 1 0 -7M and 10 -5M alendronate respectively, while the other was the con trol group without alendronate in the medium. After 4 days of adipogenic inducem ent, the adipocytes were collected by handling of 0125% trypsinase and deposit ed in 29℃ for Western-Blot analysis. Result The OD value of Western Blot band in the control group, 1 ×10 -7mol/L group and 1×10 -5mol/L group was 08098 ±01042, 05531±00534 and 04125±00354 respectively. Along with the ri se in the concentration of ALN, the expressions of leptin in the two experimenta l groups were dramatically lower than that of the control group (P<001). Conclusion The results suggest that alentronate can influence the function of intramedullary adipocytes by inhibiting leptin expression. 〔