Wen Chaoyang, Yang Xiaoli, Yan Zhifeng, Zhao Man, Yue Xiang, Cheng Xiaozhong, Zheng Zirui, Guan Kai, Dou Jianping, Xu Tao, Zhang Yanhong, Song Ting, Wei Congwen, Zhong Hui
Hainan Branch of Chinese PLA General Hospital Sanya city, Hainan 572000, China; Beijing Institute of Biotechnology, Beijing 100850, China.
Int J Cardiol. 2013 Oct 3;168(3):2242-7. doi: 10.1016/j.ijcard.2013.01.211. Epub 2013 Feb 27.
The calcification of blood vessels correlates with increased morbidity and mortality in patients with atherosclerosis, diabetes, and end-stage kidney disease. Increased inflammasome activation has been shown to play an important role in the pathogenesis of atherosclerosis. However, the contribution of inflammasome activation on the development of vascular calcification has not been investigated.
β-Glycerophosphate (β-GP) was used as a procedure to induce extensive artery calcification in primary vascular smooth muscle cells (VSMCs). Analysis of the levels of Nalp3 inflammasome complex was performed by quantitative real-time PCR and western blotting. The effect of Nalp3 deficiency on VSMC calcification was examined after transfecting Nalp3 siRNA into cultured VSMCs.
We demonstrated for the first time that the mRNA levels of Nalp3 inflammasome complex including Nalp3, ASC and caspase1 were upregulated in calcifying VSMCs, resulting in increased IL-1β secretion. Inhibition of inflammasome activation by Nalp3 RNA interference reduced IL-1β secretion and inhibited VSMC calcification. Further analysis of clinical popliteal artery specimens showed an upregulation of inflammasome complex mRNA levels (4/5) and caspase1 activity (5/5) compared with their non-calcified adjacent tissues, indicating that Nalp3 inflammasome was tightly correlated with arterial calcification disease.
Our findings indicate that activation of the Nalp3-mediated inflammatory response pathway is an important venue associated with host response and pathogenesis of VSMC calcification.
血管钙化与动脉粥样硬化、糖尿病和终末期肾病患者的发病率和死亡率增加相关。炎症小体激活增加已被证明在动脉粥样硬化的发病机制中起重要作用。然而，炎症小体激活对血管钙化发展的贡献尚未得到研究。
使用β-甘油磷酸（β-GP）诱导原代血管平滑肌细胞（VSMC）广泛的动脉钙化。通过定量实时PCR和蛋白质印迹分析Nalp3炎症小体复合物的水平。将Nalp3 siRNA转染到培养的VSMC中后，检测Nalp3缺乏对VSMC钙化的影响。
我们首次证明，在钙化的VSMC中，包括Nalp3、ASC和caspase1在内的Nalp3炎症小体复合物的mRNA水平上调，导致IL-1β分泌增加。Nalp3 RNA干扰抑制炎症小体激活可减少IL-1β分泌并抑制VSMC钙化。对临床腘动脉标本的进一步分析显示，与未钙化的相邻组织相比，炎症小体复合物mRNA水平（4/5）和caspase1活性（5/5）上调，表明Nalp-3炎症小体与动脉钙化疾病密切相关。
我们的研究结果表明，Nalp3介导的炎症反应途径的激活是与VSMC钙化的宿主反应和发病机制相关的重要环节。
