Zhao Yihan, Gu Liang, Chen Yunxuan, Lin Yibei, Xing Jincheng, Xu Diyan, Su Zhen, Huang Zhouqing
The Key Laboratory of Cardiovascular Disease of Wenzhou, Department of Cardiology, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, Zhejiang, China.
Department of Cardiology, Sichuan Academy of Medical Sciences & Sichuan Provincial People's Hospital, School of Medicine, University of Electronic Science and Technology of China, Chengdu, Sichuan, China.
Cardiovasc Ther. 2025 Feb 27;2025:6813183. doi: 10.1155/cdr/6813183. eCollection 2025.
With the progression of chronic kidney disease (CKD), we can often observe cardiac remodeling, fibrosis, and cardiac failure in patients. Cysteine-rich protein 61 (CCN1) is an extracellular matrix protein that plays a reuse role in cardiac remodeling. However, whether CCN1 participates in the crosslink between the heart and kidney in CKD and the potential mechanism remains unknown. We constructed a mouse model of CKD by 5/6 nephrectomy (5/6 Nx). Hematoxylin-eosin staining (H&E), Masson's trichrome staining, and Sirius red staining were used to observe cardiac morphology and fibrosis. H9c2 cells were treated with si-CCN1 or si-NC or mitogen-activated protein kinase (MAPK)-related inhibitors or agonist before being cultured with 5/6 Nx mouse serum. The relative protein level was detected by Western blotting. We observed that CCN1 expression was markedly enhanced in the serum and heart tissues, accompanied by disordered myocardial arrangement, obvious cardiac fibrosis, hypertrophy, and decreased cardiac systolic function reflected by echocardiography. The relative markers collagen 1 (COL-1), transforming growth factor- (TGF-), heavy-chain cardiac myosin (MyHC), and atrial natriuretic peptide (ANP) presented an increase in expression. In vivo and in vitro, after the knockdown of CCN1, the above results in the CKD group or CKD serum group were reversed; in addition, the MAPK signaling pathway was obviously activated due to 5/6 Nx, which was abolished by CCN1 inhibition. CCN1 silencing or MAPK pathway inhibition also decreased the expression of myocardial fibrosis and hypertrophy markers in H9c2 cells, while MAPK-related agonist partly reversed the effect of CCN1 inhibition. Our in vivo and in vitro study showed that specific CCN1 deficiency markedly alleviated cardiac remodeling in 5/6 Nx mice through the inhibition of the MAPK pathway.
随着慢性肾脏病（CKD）的进展，我们常常可以观察到患者出现心脏重塑、纤维化和心力衰竭。富含半胱氨酸的蛋白61（CCN1）是一种细胞外基质蛋白，在心脏重塑中发挥重要作用。然而，CCN1是否参与CKD中心脏与肾脏之间的交联以及潜在机制仍不清楚。我们通过5/6肾切除术（5/6 Nx）构建了CKD小鼠模型。采用苏木精-伊红染色（H&E）、Masson三色染色和天狼星红染色观察心脏形态和纤维化。H9c2细胞在用5/6 Nx小鼠血清培养前，用si-CCN1或si-NC或丝裂原活化蛋白激酶（MAPK）相关抑制剂或激动剂处理。通过蛋白质免疫印迹法检测相关蛋白水平。我们观察到，血清和心脏组织中CCN1表达明显增强，同时伴有心肌排列紊乱、明显的心脏纤维化、肥大，以及超声心动图显示的心脏收缩功能下降。相关标志物Ⅰ型胶原（COL-1）、转化生长因子-β（TGF-β）、心肌重链肌球蛋白（MyHC）和心房钠尿肽（ANP）的表达增加。在体内和体外，CCN1基因敲低后，CKD组或CKD血清组的上述结果得到逆转；此外，5/6 Nx明显激活了MAPK信号通路，而CCN1抑制可消除该激活作用。CCN1沉默或MAPK通路抑制也降低了H9c2细胞中心肌纤维化和肥大标志物的表达，而MAPK相关激动剂部分逆转了CCN1抑制的作用。我们的体内和体外研究表明，特异性CCN1缺乏通过抑制MAPK通路显著减轻了5/6 Nx小鼠的心脏重塑。
