系统性红斑狼疮模型鼠骨 BMP/Smads信号通路状态的研究
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国家自然科学基金(81202358);镇江市社会发展项目(SH2013028);2013年镇江市学术技术带头人专项基金;江苏省“六大人才高峰”项目(WSN-039)


Status of BMP/Smads signaling on the bone from mouse model of systemic lupus erythematosus
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    摘要:

    目的:探讨狼疮鼠(MRL/lpr)骨BMP/Smads信号通路表达情况。方法:分离小鼠股骨制备组织切片,常规苏木素-伊红(HE)染色,观察骨质情况;免疫组化观察骨组织中骨形态发生蛋白-2(bone morphogenetic protein-2,BMP-2)表达;密度梯度离心法和贴壁筛选法分离培养骨髓间充质干细胞 (bone marrow masenchymal stem cells,BMMSCs),细胞爬片后免疫荧光法观察BMP-2-p-Smad1/5/8蛋白表达情况;BMP-2诱导BMMSCs向成骨细胞分化,RT-PCR法检测BMMSCs ALP-Runx2基因mRNA水平,ALP染色鉴定早期成骨情况。结果:狼疮鼠骨皮质较正常鼠减少,皮质骨占骨体积比例较正常鼠减低(P < 0.01);狼疮鼠股骨BMP-2表达与正常鼠无明显差别(P > 0.05);细胞免疫荧光显示狼疮鼠BMMSCs BMP-2-p-Smad1/5/8表达减弱(P < 0.05);狼疮鼠BMMSCs BMP-2刺激7 d后ALP活性较正常鼠减低,BMP-2刺激3 d后ALP mRNA水平较正常鼠减弱(P < 0.01),Runx2 mRNA水平较正常鼠无明显差别(P > 0.05)。结论:狼疮鼠骨 BMP/Smads 信号通路处于抑制状态。

    Abstract:

    Objective:To investigate the status of BMP/Smads signaling pathway on the bone of lupus mice(MRL/lpr). Methods:The femurs of the mice were isolated,and the bone structure was detected by hematoxylin-eosn(HE)staining. The protein expression levels of bone morphogenetic protein-2 (BMP-2)were measured by immunohistochemistry. Bone marrow mesenchymal stem cells(BMMSCs)were isolated by the density gradient centrifugation and the adherence screening methods. After BMMSCs growing on coverslips,the protein of BMP-2 and PSmad1/5/8 was valued by immumofluorescence method. The mRNA expressions of ALP,Runx2 of BMMSCs were measured by Real-time PCR,and the Alkaline phosphatase (ALP)staining was performed for measuring the early osteogenic differentiation induced by BMP-2. Results:Compared with the controlled mice,the cortex of MRL/lpr mice were reduced (P < 0.01). There was no difference in the BMP-2 expression on the bone of the two groups by immunohistochemistry detection(P > 0.05). The expressions of BMP-2 and PSmad1/5/8 in BMMSCs of lupus mice were lower than those of the control group(P < 0.05). After 7 days of BMP-2 stimulation,ALP activities of BMMSCs from lupus mice were decreased compared with the control group. The mRNA levels of ALP in lupus mice were lower than that of C3He/HeJ mice (P < 0.01),while there were no differences in the Runx2 mRNA levels between the two groups after BMP-2 stimulation(P > 0.05). Conclusion:BMP/Smads signaling was inhibited in the bone of lupus mouse.

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宋冬明,崔 婷,裘影影,芮金兵,费小明,徐欣欣,李 晶,汤 郁.系统性红斑狼疮模型鼠骨 BMP/Smads信号通路状态的研究[J].南京医科大学学报(自然科学版),2015,(3):299-303-314

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  • 收稿日期:2014-07-07
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  • 在线发布日期: 2015-03-10
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