激素对人骨髓间充质干细胞神经肽受体mRNA表达的影响  

Rffect of steroid on mRNA expression of neuropeptide receptors in human bone marrow mesenchy- mal stem cells

作　　者：王义生[1] 张战峰[1] 李月白[2] 赵国强[3] 朱绍阳[1] 

机构地区：[1]郑州大学第一附属医院骨科河南省高等学校临床医学重点学科开放实验室,450052 [2]郑州大学基础医学院生物化学与分子生物学教研室 [3]郑州大学基础医学院微生物与免疫学教研室

出　　处：《中华实验外科杂志》2012年第5期933-936,共4页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金资助项目（81171776）

摘　　要：目的观察激素对人骨髓间充质干细胞（hBMSCs）内神经肽受体降钙素基因相关肽受体（CGRPR）、P物质受体（SPR）及过氧化物酶体增殖子活化受体1（PPA脚）、成骨转录因子Runx2等基因表达的影响。方法密度梯度离心联合贴壁分选法体外分离培养hBMSCs，流式细胞仪鉴定后传至第3代，随机分为两组，实验组细胞给予1×10^-7mol／L地塞米松干预9d，对照组正常培养。观察细胞形态、生长趋势，油红O染色脂肪细胞，实时定量-聚合酶链反应（RT—qPCR）分析mRNA表达的变化。结果分离培养的贴壁细胞呈纤维状、漩涡样生长，CD29、CD44表达率为（91．4±0．7）％、（93．8±0．5）％，CD34表达率为（2．7±0．8）％。实验组细胞生长缓慢、出现大量脂肪细胞，对照组细胞增殖旺盛、未见脂肪细胞。RT—qPCR发现实验组CGRPRmRNA、SPRmRNA、Runx2mRNA的表达显著降低，2△△^Cr分别为0．10±0．03、0．16±0．04、0．23±0．05（P均〈0．01）。实验组PPARlmRNA表达量2△△^Cr是对照组的（5．08±3．37）倍，差异有统计学意义（P〈0．01）。结论激素能够诱导hBMSCs成脂分化，下调细胞中CGRPRmRNA、SPRmRNA、Runx2mRNA表达，抑制细胞成骨分化与生长增殖，使骨修复能力不足，这可能与激素性骨坏死的发生机制有关。Objective To investigate the effect of steroid on the mRNA expression of calcitonin gene-related peptide receptor （CGRPR）, substance P receptor （SPR）, peroxisome proliferators-activated receptor-γ （PPARγ） , and Runx2 in human bone marrow mesenchymal stem cells （hBMSCs）. Methods hBMSCs were isolated and purified by density gradient centrifugation combined with plastic-adhering culture, and identified by flow cytometry. At the third generation, hBMSCs were randomly divided into two groups： experimental group and control group. The hBMSCs in experimental group were treated with 1×10^-7mol/L dexamethasone, and those in control group were routinely cultured. The morphological changes and growth trend wer observed, and adipoeytes were stained with oil red O. The expression changes of mRNA were analyzed by using real-time quantitative reverse transeription-polymerase chain reaction （RT-qPCR）. Results Adherent cells were fiber-shaped and had a swirl-like growth. The positive rate of CD29 and CD44 was （91.4±0.7）% and （93.8±0.5）%, while that of CD34 was （2.7±0.8）%, respectively. The cells in experimental group grew slower than in control group, while those in control group proliferated greatly. The number of adipocytes in control group was increased significantly. The mRNA expression levels of CGRPR, SPR and Runx2 in experimental group was significantlly lower than those in control group, with the 2△△^Cr being 0. 10±0.03, 0. 16±0. 04, and 0. 23±0.05 respectively （all P 〈 0. 01 ）, while the 2△△^Cr of PPARγ was 5.08±3.37 （ P 〈 0.01）. Conclusion Steroid could induce hBMSCs adipogenic differentiation, inhibit the mRNA expression of CGRPR, SPR, Runx2, and then restrain cells growth, proliferative ability and osteogenic differentiation of the hBMSCs, which might be related to the pathogenesis of steroid-induced osteonecrosis.

关 键 词：骨髓间充质干细胞 激素 神经肽 降钙素基因相关肽受体 P物质受体 

分 类 号：R681[医药卫生—骨科学]