不同浓度瘦素对乳鼠成骨细胞增殖及VEGF mRNA表达的影响

Different Concentrations of Leptin Acts on Osteoblast Proliferation and Expression of VEGF mRNA

目的通过观察不同浓度瘦素对离体培养乳鼠颅骨成骨细胞的作用,探讨瘦素(Leptin)对成骨细胞增殖以及成骨细胞上血管内皮细胞生长因子(vascular endothelial growth factor,VEGF)表达的影响,为促进骨愈合过程中骨修复、再生的临床治疗提供一定的理论依据。方法取新生24h乳鼠颅盖骨,经酶解消化后得到原代成骨细胞,进行体外培养并鉴定。采用MTT法(噻唑蓝染色法)测定各组的OD值,观察瘦素对成骨细胞增殖能力的影响;采用RT-PCR法测定瘦素作用成骨细胞后VEGF mRNA的表达情况。结果经细胞鉴定后证实其系成骨细胞;经MTT法检测表明,瘦素作用24h后,各实验组OD值随瘦素浓度升高而增加;经RT-PCR法检测表明,VEGF mRNA可在乳鼠成骨细胞内稳定表达;半定量分析表明,不同瘦素剂量组间VEGF mRNA的表达量呈现一定规律,随着瘦素浓度的升高,VEGF mRNA的表达量逐渐增加。结论瘦素可促进成骨细胞增值;VEGF mRNA在成骨细胞内稳定表达;瘦素可上调成骨细胞上VEGF mRNA的表达,并存在一定的剂量依赖关系。

Objective To observe the effect of leptin on osteoblasts proliferation and vascular endothelial growth factor （VEGF） expression. To provide a theoretical basis for the clinical treatment of bone repair and regeneration after injury. Methods Primary osteoblasts were cultured in vitro by enzymatic digestion. OD values in each group was measured by MTT method;the expression of VEGF mRNA was assessed by RT-PCR. Results The MTT showed that after 24h, OD values in all experimental groups rises with the increasing concentration of leptin;The RT-PCR showed that, VEGF mRNA in neonatal rat osteoblasts has a stable expression; Semi-quantitative analysis showed that with the rising of leptin concentrations, VEGF mRNA expression increased gradually. Conclusions Leptin can promote osteoblast proliferation; VEGF mRNA can express stability in osteoblasts; VEGF mRNA expression could be up-regulated by leptin in a dose-dependent manner.