关键词: 瘦素, 骨髓基质细胞, 基因转染, 腺病毒, β磷酸三钙

Abstract: Objective To investigate the effects of human leptin (hLEP) gene transfection on rat bone marrow stromal cells (rBMSCs). Methods rBMSCs were cultured and transfected with adenoviruses encoding hLEP (Ad5-hLEP-EGFP) in vitro as experimental group while rBMSCs transfected with Ad5-EGFP and non-transfected were control groups. The proliferation was detected by MTT and the expression of collagen type ? (Col-?) and alkaline phosphatase (ALP) were assessed by real-time PCR. The ability of mineralized nodule forming was also compared by Alizarin red staining. What’s more, the combination of transfected rBMSCs and β-tricalcium phosphate (β-TCP) was constructed and the osteogenic ability of the construction was evaluated in nude mice. Results hLEP could be transfected into rBMSCs successfully by adenovirous. After transfection, the proliferation was not affected while Col-? and ALP expressions were more pronounced in rBMSCs transfected with Ad5-hLEP-EGFP (P < 0.05). Alizarin red staining showed the ability of mineralized nodule forming was also up-regulated in Ad5-hLEP-EGFP group (P < 0.05). In addition, the transfected rBMSCs could adhere to β-TCP and survived well and the combination showed more new bone like tissue formation in nude mice compared to control groups. Conclusions rBMSCs transfected with hLEP may have potentials to be used in bone or periodontal tissue regeneration.

Key words: leptin, bone marrow stromal cells, gene transfection, adenovirus, β-tricalcium phosphate