C-type natriuretic peptide inhibiting vascular calcification might involve decreasing bone morphogenic protein 2 and osteopontin levels
C-type natriuretic peptide inhibiting vascular calcification might involve decreasing bone morphogenic protein 2 and osteopontin levels
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DOI：
10.1007/s11010-014-2019-1
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发表时间：
2014-07-01
影响因子：
4.3
通讯作者：
Jiang, Zhi-Sheng
中科院分区：
文献类型：
--
作者：
Chen, Jing-Jing;Zhang, Jing;Jiang, Zhi-Sheng
血管钙化（VC）与晚期慢性肾病患者的发病率和死亡率升高高度相关。血管活性肽等旁分泌/自分泌因子参与了VC的发展。在这里，我们研究了新型肽c型利钠肽（CNP）在血管中的表达，测试了其体内和体外预防VC的能力，并探讨了其机制。用维生素D3 +尼古丁（VDN）诱导大鼠主动脉VC。微渗透泵给药500 ng/kg/h。von Kossa染色检查钙化；采用放射免疫法检测主动脉和钙化血管平滑肌细胞（VSMCs）中CNP和环鸟苷单磷酸（cGMP）含量，采用实时荧光定量PCR和Western blot检测mRNA和蛋白水平。vdn处理大鼠主动脉CNP含量升高，其受体钠肽受体B表达降低，血管钙沉积增加，碱性磷酸酶（ALP）活性增加。与载药处理相比，低CNP水平的vdn处理大鼠血管钙沉积和ALP活性增加，这在培养的VSMCs中得到进一步证实。与对照组相比，给予CNP可显著降低vdn处理主动脉的VC，这在钙化的VSMCs中得到证实。体外CNP可改善α -肌动蛋白表达的下降。此外，骨桥蛋白（osteopontin， OPN）在钙化主动脉中的表达水平显著上调，而CNP增加了钙化主动脉中OPN的表达。此外，CNP下调钙化主动脉和VSMCs中OPN和骨形态发生蛋白2 （BMP-2）的表达。蛋白激酶G抑制剂H7显著阻断了CNP对OPN和BMP-2表达的调节以及CNP对钙化VSMCs的有益作用。局部内源性CNP及其受体系统受损可能与体内VC大鼠主动脉矿化增加有关，并且CNP至少在一定程度上通过cGMP/PKG途径抑制体内和体外VC的发育。
Vascular calcification (VC) is highly associated with increased morbidity and mortality in patients with advanced chronic kidney disease. Paracrine/autocrine factors such as vasoactive peptides are involved in VC development. Here, we investigated the expression of the novel peptide C-type natriuretic peptide (CNP) in the vasculature, tested its ability to prevent VC in vivo and in vitro, and examined the mechanism involved. Rat aortic VC was induced by vitamin D3 plus nicotine (VDN). CNP (500 ng/kg/h) was administered by mini-osmotic pump. Calcification was examined by von Kossa staining; CNP and cyclic guanosine monophosphate (cGMP) contents were detected by radioimmunoassay, and mRNA and protein levels were examined by real-time PCR and Western blot analysis in aortas and calcified vascular smooth muscle cells (VSMCs). VDN-treated rat aortas showed higher CNP content and decreased expression of its receptor natriuretic peptide receptor B, along with increased vascular calcium deposition and alkaline phosphatase (ALP) activity. Low CNP levels were accompanied by increased vascular calcium deposition and ALP activity in VDN-treated rats when compared to vehicle treatment, which was further confirmed in cultured VSMCs. Administration of CNP greatly reduced VC in VDN-treated aortas compared with controls, which was confirmed in calcified VSMCs. The decrease in alpha-actin expression was ameliorated by CNP in vitro. Moreover, protein expression levels of osteopontin (OPN) were significantly up-regulated in calcified aortas, and CNP increased OPN expression in calcified aortas. Furthermore, CNP downregulated OPN and bone morphogenic protein 2 (BMP-2) expression in calcified aortas and VSMCs. Modulation of OPN and BMP-2 expression by CNP and the beneficial effects of CNP on calcified VSMCs were blocked significantly by protein kinase G inhibitor H7. Impaired local endogenous CNP and its receptor system may be associated with increased mineralization in vivo in rat aortas with VC, and administration of CNP inhibits VC development in vivo and in vitro, at least in part, via a cGMP/PKG pathway.