酸性环境对慢性肾功能衰竭大鼠血管中膜钙化的影响及其机制
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(河北医科大学第四医院肾内科, 河北省石家庄市 050011)

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张胜雷,硕士,主治医师,研究方向为慢性肾脏病血管钙化,E-mail为lei06352511@126.com。

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河北省自然科学基金资助项目(H2012206157)


Effect and Mechanism of Acid Environment on Media Vascular Calcification in Rats with Chronic Renal Failure
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Department of Nephrology, the Forth Hospital of Hebei Medical University, Shijiazhuang, Hebei 050011, China)

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    摘要:

    目的 探讨酸性环境对慢性肾功能衰竭大鼠血管中膜钙化的影响及可能的作用机制。方法 体内实验:将雄性SD大鼠随机分为对照组、肾功能衰竭组、血管钙化组和酸干预组,造模成功后采用von Kossa染色和邻甲酚酞络合酮比色法测定大鼠胸主动脉血管钙化情况。体外实验:分离培养SD大鼠胸主动脉血管平滑肌细胞(VSMC),随机分为正常对照组、高磷组、酸干预组和抑制剂组(BMP信号通路抑制剂Noggin),采用茜素红染色和邻甲酚酞络合酮比色法检测细胞钙化情况,酶联免疫吸附法检测细胞碱性磷酸酶(ALP)活性,RT-PCR和Western blot检测细胞BMP-2、Smad1、Runx2的表达情况。结果 体内实验:成功制备了慢性肾功能衰竭大鼠血管钙化模型;与血管钙化组相比,酸干预组大鼠血管钙盐沉积明显减轻(P<0.05)。体外实验:与正常对照组相比,高磷组细胞钙盐沉积明显增加(P<0.05),ALP活性和Runx2表达均增高(P<0.05);与高磷组相比,酸干预组细胞钙盐沉积减少(P<0.05),ALP活性、Runx2、BMP-2和Smad1表达均降低(P<0.05);与高磷组相比,抑制剂组细胞钙盐沉积和Runx2表达均降低(P<0.05)。结论 酸性环境可以抑制慢性肾功能衰竭大鼠血管中膜钙化的发生,其可能的机制之一是通过BMP-2信号通路抑制了VSMC成骨/成软骨样表型转化来实现的。

    Abstract:

    Aim To explore the effect and mechanism of acid environment on media vascular calcification of chronic renal failure rats. Methods In vivo experiment:healthy male SD rats were randomly divided into control group, renal failure group, vascular calcification group and acid intervention group, calcification was measured by Alizarin red staining and enzyme linked immunosorbent assay. In vitro experiment:vascular smooth muscle cells (VSMC) were primarily cultured in vitro and randomly divided into control group, high phosphorus group, acid intervention group and inhibitor group, calcium deposition and ALP activity were measured by Alizarin red staining, quantification of calcium and enzyme linked immunosorbent assay, the expression of BMP-2, Smad1 and Runx2 was detected by RT-PCR and Western blot. Results In vivo experiment:the vascular calcification model of rats with chronic renal failure was successfully prepared. Compared with vascular calcification group, calcium deposition in acid intervention group decreased (P<0.05).In vitro experiment:compared with control group, calcium deposition, alkaline phosphatase (ALP) activity and the expression of Runx2 in high phosphorus group increased (P<0.05). Compared with high phosphorus group, calcium deposition, ALP activity and the expression of Runx-2, BMP-2 and Smad1 in acid intervention group decreased (P<0.05). Compared with high phosphorus group, calcium deposition and the expression of Runx2 in inhibitor group also decreased (P<0.05). Conclusions Acid environment can inhibit the occurrence of vascular calcification in chronic renal failure rats, the mechanism was presumably that vascular smooth muscle cells osteogenesis/chondrogenesis-like phenotypic transformation was inhibited by BMP-2 signaling pathway.

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张胜雷,徐金升,郭利萍,白亚玲,张俊霞,崔立文,张慧然.酸性环境对慢性肾功能衰竭大鼠血管中膜钙化的影响及其机制[J].中国动脉硬化杂志,2016,24(12):1201~1206.

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  • 收稿日期:2016-03-17
  • 最后修改日期:2016-05-11
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  • 在线发布日期: 2017-02-09