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| 淫羊藿苷促进大鼠骨髓基质细胞成骨性分化过程中对 p-AKT及eNOS、iNOS表达量的影响 |
| The expression of p-AKT，eNOS and iNOS in the process of ICA promotes osteogenic differentiation of BMSCs |
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| DOI： |
| 中文关键词: 大鼠骨髓基质细胞 淫羊藿苷 NOS AKT |
| 英文关键词:Rat bone marrow stromal cells Icariin Nitric oxide synthase AKT |
| 基金项目:甘肃省科技重大专项资助项目（09ZNKDA025 )、国 家自然科学基金（81270963) |
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| 中文摘要: |
| 目的研究淫羊藿苷(Icariin. ICA)促进体外培养大鼠骨髓基质细胞（Bone marrow stromal cells . BMSCs)的成骨性分化 机理。方法贴壁分离法培养大鼠骨髓基质细胞并用流式检测细胞纯度。骨髓基质细胞传代至第2代时分别用含有成骨性 诱导剂的 1x10_5 mol/L ICA 和 LY294002 处理 24 h 后用 Real~time RT-PCR 检测 ALP、eNOS 和 iNOS 的 mRNA 表达水平， Western blotting检测ALP、p4KT、eNOS和iNOS的蛋白表达量。结果原代骨髓基质细胞体外培养传代至第2代时纯度可达 95%以上。基因和蛋白质检测均显示ICA可极显著的促进碱性磷酸酶（ALP)的表达（P <0.01), PI3K的特异性抑制剂 LY294002会抑制ICA对ALP的提高,ICA + LY294002组与CON组的表达差别不明显。p^KT的蛋白表达变化与ALP相似。 ICA可以提高eNOS表达(P <0. 01)，加抑制剂能使其降低（P <0. 01),而iNOS的ICA组与ICA + LY294002组无明显差别。 结论ICA促进BMSCs成骨性分化过程涉及到PI3K/AKT通路，且AKT位于eNOS的上游;eNOS和iNOS共同作用，但iNOS 起主要作用。 |
| 英文摘要: |
| Objective To investigate the effect mechanism of Icariin (ICA) on osteogenic differentiation of bone marrow stromal cells (BMSCs). Methods Rat BMSCs were isolated according to plastic adherence and identified by flow cytometry . The mRNA levels of alkaline phosphatase (ALP)，eNOS，and iNOS were detected using quantitative real -time RT-PCR. The protein levels of ALP， p- AKT，eNOS，and iNOS were detected using by Western blotting. Results More than 95% of the 2nd passage rat BMSCs were immune positive，with less than 5% were immune negative. The result showed that 10 5 moL/L ICA could significantly promote the expression of ALP (P <0.01)，while the expression in LY294002 group was lower than that in the ICA group (P < 0. 01). The expression in ICA + LY294002 group and control group had no significant difference . The protein levels of p -A.KT were similar to ALP. The expression of eNOS in ICA group increased (P < 0. 01)，while decreased in LY294002 group ( P < 0. 01). However， no significant difference of protein level was observed between ICA group and ICA + LY294002 group. Conclusion ICA promotes osteogenic differentiation of BMSCs，and the PI3K/AKT signal pathway is involved in this process. AKT is located at the upstream of eNOS. eNOS and iNOS both can affect the process，while iNOS may play a major role. |
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