血清BMP-2、BMP-4检测在慢性肾脏病大鼠主动脉钙化中的意义
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(西南医科大学附属医院肾病内科,四川省泸州市 646000)

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魏晓,现工作于四川省成都市温江区人民医院,E-mail为757061143@qq.com。通信作者欧三桃,医学博士,教授,E-mail为ousantao@163.com。

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Significance of serum BMP-2 and BMP-4 detection in aortic calcification in rats with chronic kidney disease
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Department of Nephrology, the Affiliated Hospital of Southwest Medial University, Luzhou, Sichuan 646000, China)

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    摘要:

    目的 建立慢性肾脏病(CKD)大鼠主动脉钙化模型,探讨血清骨形态发生蛋白2(BMP-2)、骨形态发生蛋白4(BMP-4)与主动脉钙化程度的相关性。方法 55只SD大鼠随机分为对照组(20只)和CKD组(35只,由于预实验中CKD组大鼠后期存在死亡现象,为保证实验过程每个时间的实验样本量充足,因此CKD组样本量较对照组多),分别于第2、4、6、8周收集血液、尿液、主动脉、肾脏;肾脏HE染色评估肾脏病理改变;茜素红染色法、钙含量测定评估主动脉钙化程度;ELISA测定大鼠血清BMP-2、BMP-4水平;免疫组织化学检测主动脉BMP-2、BMP-4蛋白表达。结果 与对照组各时间点相比,CKD组大鼠24 h尿蛋白定量、血清尿素氮、肌酐、胱抑素C水平明显增加(P<0.05)。肾脏HE染色显示,自第2周起,肾小管出现扩张,小管内棕黄色物质沉积,随时间进展,逐渐出现间质纤维化,肾小球部分萎缩,血管减少,炎性细胞浸润。茜素红染色随时间进展,第8周出现大量橘红色颗粒物沉积伴钙结节形成。CKD组钙含量明显增加且与血清BMP-2 (r=0.929,P<0.01) 及BMP-4 (r=0.702,P<0.01)明显正相关。同时,自模型成功建立第4周,主动脉BMP-2、BMP-4蛋白表达即明显增加。结论 血清BMP-2、BMP-4表达增加可作为CKD血管钙化的血清标志物。

    Abstract:

    Aim To investigate the association between serum bone morphogenetic protein-2 (BMP-2), bone morphogenetic protein-4 (BMP-4) levels and the severity of vascular calcification in CKD rat model. Methods Fifty-five SD rats were randomly divided into control group (n=20) and CKD group (n=35). The rats were sacrificed at 2,4, 6,8 week respectively. The kidney and aortic pathologies were analyzed. Blood biochemical indicators, serum BMP-2 and BMP-4 levels, and aortic calcium content were determined. The expression levels of BMP-2, BMP-4 were examined by immunohistochemistry. Results Compared with control group, the CKD rats exhibited a significantly increase in 24 h urinary protein, serum urea nitrogen, creatinine and cystatin C (P<0.05). Kidney HE staining showed that renal tubular expansion began to emerge since the second week, accompanying with brown yellow substance deposition, interstitial fibrosis, glomerular partial atrophy, and inflammatory cells infiltration. Alizarin red staining showed massive granular deposition and formation of calcified nodules in aorta at the 8th week. The aortic calcium content was significantly increased, which was positively correlated with serum BMP-2 (r=0.929, P<0.01) and serum BMP-4 (r=0.702, P<0.01) levels in CKD rats. The expression of BMP-2 and BMP-4 in aorta increased significantly after successful establishment of the model at the fourth week. Conclusion Elevated serum BMP-2 and BMP-4 levels may serve as serum markers for CKD vascular calcification.

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魏晓,侯静,吴蔚桦,刘琦,李莹,龚财判,李丽,欧三桃.血清BMP-2、BMP-4检测在慢性肾脏病大鼠主动脉钙化中的意义[J].中国动脉硬化杂志,2018,26(9):900~905.

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  • 收稿日期:2017-11-10
  • 最后修改日期:2018-02-12
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  • 在线发布日期: 2018-10-16