Overexpression of C1q/Tumor Necrosis Factor–Related Protein-3 Promotes Phosphate-Induced Vascular Smooth Muscle Cell Calcification Both In Vivo and In Vitro
Overexpression of C1q/Tumor Necrosis Factor–Related Protein-3 Promotes Phosphate-Induced Vascular Smooth Muscle Cell Calcification Both In Vivo and In Vitro
复制标题
DOI：
10.1161/atvbaha.114.303301
复制
发表时间：
2014-05
期刊：
影响因子：
--
通讯作者：
Yun-jiang Zhou;Jin-yu Wang;H. Feng;Cheng Wang;Li Li-Li;Dan Wu;Hong Lei;Hao Li;Li-Ling Wu
中科院分区：
文献类型：
--
作者：
Yun-jiang Zhou;Jin-yu Wang;H. Feng;Cheng Wang;Li Li-Li;Dan Wu;Hong Lei;Hao Li;Li-Ling Wu
目的：血管钙化与心血管疾病发病率和死亡率升高高度相关。C1q/肿瘤坏死因子相关蛋白-3 （CTRP3）是一种新发现的脂肪因子，在心血管系统中起重要作用。在这里，我们研究了CTRP3在血管钙化中的作用及其潜在机制。方法与结果：采用腺嘌呤诱导的大鼠慢性肾衰竭模型，模拟动脉内侧钙化过程。慢性肾衰竭大鼠血清及腹主动脉中CTRP3水平升高。膜周基因递送CTRP3显著加速腹主动脉和动脉环的钙化。在培养的血管平滑肌细胞（VSMCs）中，CTRP3增加了甘油磷酸酯诱导的钙沉积和碱性磷酸酶活性。虽然CTRP3本身不足以诱导VSMCs的钙化，但它上调了成骨标志基因的表达，包括矮子相关转录因子2 （Runx2）、骨形态发生蛋白2和骨桥蛋白。CTRP3进一步增强-甘油磷酸盐诱导的平滑肌-肌动蛋白和平滑肌22的下调，同时增加-甘油磷酸盐诱导的VSMCs中成骨标志物的表达。相比之下，VSMCs中CTRP3的下调可有效抑制-甘油磷酸酯诱导的钙化。在机制上，Runx2的下调抑制了ctrp3促进的VSMC钙化。CTRP3增加细胞外信号调节的激酶1/2磷酸化和活性氧的产生。与细胞外信号调节激酶1/2上游激酶抑制剂U0126预孵育对ctrp3诱导的活性氧产生没有影响。然而，用n -乙酰- l-半胱氨酸（一种活性氧清除剂）预处理可以抑制ctrp3诱导的细胞外信号调节激酶1/2磷酸化。n -乙酰- l-半胱氨酸和U0126均能显著抑制ctrp3诱导的Runx2上调和钙化结节形成。结论- ctrp3通过活性氧-胞外信号调节激酶1/2-Runx2通路，通过增强磷酸盐诱导的VSMC成骨转变，促进血管钙化。
Objective—Vascular calcification is highly correlated with increased cardiovascular morbidity and mortality. C1q/tumor necrosis factor–related protein-3 (CTRP3) is a newly identified adipokine that plays important roles in cardiovascular system. Here, we investigated the role of CTRP3 in vascular calcification and its underlying mechanism. Approach and Results—Adenine-induced chronic renal failure rat model was used to mimic the process of arterial medial calcification. The level of CTRP3 was elevated in serum and abdominal aorta of chronic renal failure rats. Periadventitial gene delivery of CTRP3 significantly accelerated the calcification of abdominal aorta and arterial ring. In cultured vascular smooth muscle cells (VSMCs), CTRP3 increased -glycerophosphate–induced calcium deposition and alkaline phosphatase activity. Although CTRP3 alone was not sufficient to induce calcification in VSMCs, it upregulated the expression of osteogenic marker genes including runt-related transcription factor 2 (Runx2), bone morphogenetic protein 2, and osteopontin. CTRP3 further enhanced -glycerophosphate–induced downregulation of smooth muscle -actin and smooth muscle 22, while augmenting osteogenic marker expression in VSMCs induced by -glycerophosphate. In contrast, knockdown of CTRP3 in VSMCs potently suppressed -glycerophosphate–induced calcification. Mechanistically, knockdown of Runx2 inhibited CTRP3-promoted VSMC calcification. CTRP3 increased extracellular signal–regulated kinase 1/2 phosphorylation and reactive oxygen species production. Preincubation with U0126, an extracellular signal–regulated kinase 1/2 upstream kinase inhibitor, had no effect on CTRP3-induced reactive oxygen species production. However, pretreatment with N-acetyl-L-cysteine, a reactive oxygen species scavenger, suppressed CTRP3-induced extracellular signal–regulated kinase 1/2 phosphorylation. Both N-acetyl-L-cysteine and U0126 significantly inhibited CTRP3-induced upregulation of Runx2 and calcified nodule formation. Conclusions—CTRP3 promotes vascular calcification by enhancing phosphate-induced osteogenic transition of VSMC through reactive oxygen species–extracellular signal–regulated kinase 1/2–Runx2 pathway.