系统性红斑狼疮患者外周血BMP/Smads信号通路相关基因表达及与OPG/RANKL的相关性分析
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国家自然科学基金(81202358,81571582);江苏省六大人才高峰(WSN-039);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


Gene expressions of BMP/Smad signaling pathway in the peripheral blood of patients with systemic lupus erythematosus and the correlation analysis with OPG/RANKL
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    目的:研究系统性红斑狼疮(SLE)患者外周血BMP/Smads信号通路相关基因表达,了解其与OPG/RANKL相关性,探讨其在SLE骨代谢中的意义-方法:选取SLE患者26例及正常对照组20例,运用实时定量PCR方法检测两组外周血单个核细胞BMP/Smads信号通路相关基因BMP-2-Smad1-Smad5-Smad8及RANKL-OPG的表达,ELISA法定量检测血清中RANKL-OPG水平,计算OPG/RANKL比值;分析Smad1-Smad5-Smad8基因表达量与疾病活动相关指标及OPG/RANKL相关性-结果:SLE患者外周血单个核细胞OPG/RANKL系统中OPG基因mRNA表达水平较正常对照组减低 (0.002 5 ± 0.000 7 vs. 0.017 6 ± 0.006 7,P < 0.05),RANKL基因mRNA表达水平无统计学差异 (0.003 3 ± 0.001 0 vs. 0.002 4 ± 0.000 9,P > 0.05),SLE患者OPG/RANKL比值较正常对照组明显减低 (0.229 6 ± 0.071 2 vs. 0.609 5 ± 0.165 1,P < 0.01)-SLE患者Smad1-Smad8基因mRNA表达水平较正常对照组明显减低 (0.003 1 ± 0.000 4 vs. 0.006 6 ± 0.000 7;0.003 3 ± 0.000 5 vs. 0.005 6 ± 0.000 6,P < 0.01),Smad5基因mRNA表达较正常对照组减低(0.001 7 ± 0.000 3 vs. 0.004 2 ± 0.000 4,P < 0.05),SLE患者与正常对照组BMP-2基因mRNA表达水平无统计学差异(0.006 9 ± 0.002 0 vs. 0.010 5 ± 0.002 1,P > 0.05)-SLE患者组血清RANKL表达较正常对照组明显增高(P < 0.01),OPG表达在两者间无明显统计学意义(P > 0.05),SLE血清OPG/RANKL较正常对照组明显减低(P=0.006)-相关性分析显示,SLE患者Smad8 mRNA水平-OPG/RANKL均与血沉呈正相关 (P < 0.05)-OPG mRNA水平与Smad8 mRNA表达水平呈正相关P <0.01,而RANKL与Smad1 mRNA基因表达呈负相关(P < 0.01)-结论:SLE患者体内BMP/Smads系统基因表达存在异常,可能与SLE的骨代谢异常相关-

    Abstract:

    Objective: To detect the mRNA levels of genes of BMP/Samd signaling pathway in peripheral blood mononuclear cells of patients with systemic lupus erythematosus (SLE) and to determine their correlations with the ratio of OPG/RANKL, thus to further investigate the effect of BMP/Smad pathway on the bone metabolism of SLE patients. Methods: The mRNA levels of RANKL, OPG, BMP-2 and Smad1/5/8 gene in peripheral peripheral blood mononuclear cells from 26 SLE patients and 20 normal controls were detected by rea1-timePCR (RT-PCR), and the protein level of OPG and RANKL were measured by ELISA. The correlation analysis between these genes of BMP/Smad signaling and the ratio of OPG/RANKL or some indexes of disease activity were studied. Results: The mRNA levels of OPG decreased in SLE patients compared with normal controls (0.002 5 ± 0.000 7 vs. 0.017 6 ± 0.006 7, P < 0.05), and RANKL levels had no differences between the two groups (0.003 3 ± 0.001 0 vs. 0.002 4 ± 0.000 9, P > 0.05), which led to a decreased ratio of OPG/RANKL (0.229 6 ± 0.071 2 vs. 0.609 5 ± 0.165 1, P < 0.01). Both the mRNA levels of Smad1 and Smad8 were significantly decreased in SLE patients than those in controls (0.003 1 ± 0.000 4 vs. 0.006 6 ± 0.000 7; 0.003 3 ± 0.000 5 vs. 0.005 6 ± 0.000 6,P <0.01), and also the level of Smad5 was decreased (0.001 7 ± 0.000 3 vs. 0.004 2 ± 0.000 4, P < 0.05). There were no differences on the BMP-2 gene expression between the two groups (0.006 9 ± 0.002 0 vs. 0.010 5 ± 0.002 1,P > 0.05). Serum protein levels of RANKL were higher in SLE patients than those in controls (P < 0.01), while OPG levels had no differences between the two groups, which led to an decreased ratio of OPG/RANKL (P = 0.006) in SLE patients. Analysis showed that positive correlation between mRNA level of Smad8, ratio of OPG/RANKL and ESR levels of SLE patients (P < 0.05). Moreover, mRNA levels of Smad8 and OPG were found positively related (P < 0.01), while that of Smad1 and RANKL were negatively related (P < 0.01). Conclusion: The gene expressions of BMP/Smads signaling pathway were abnormal in SLE patients, which may be involved in the mechanism of unbalanced bone metabolism of patients.

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宋冬明,崔 婷,刘瑞霞,裘影影,芮金兵,费小明,李 晶,汤 郁.系统性红斑狼疮患者外周血BMP/Smads信号通路相关基因表达及与OPG/RANKL的相关性分析[J].南京医科大学学报(自然科学版),2016,(1):55-59

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  • 收稿日期:2015-06-23
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  • 在线发布日期: 2016-01-30
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