An Na, Chen Yu, Xing Yanfen, Wu Honghua, Gao Xiongyi, Chen Hengwen, Song Ke, Li Yuanyuan, Li Xinye, Yang Fan, Pan Xiandu, He Xiaofang, Wang Xin, Li Yang, Gao Yonghong, Xing Yanwei
Guang'anmen Hospital, Chinese Academy of Chinese Medical Sciences, Beijing 100053, China.
Key Laboratory of Chinese Internal Medicine of Ministry of Education, Dongzhimen Hospital Affiliated to Beijing University of Chinese Medicine, Beijing 100700, China.
Evid Based Complement Alternat Med. 2020 Oct 19;2020:9502651. doi: 10.1155/2020/9502651. eCollection 2020.
Previous studies have demonstrated that calcium-/calmodulin-dependent protein kinase II (CaMKII) and calcineurin A-nuclear factor of activated T-cell (CnA-NFAT) signaling pathways play key roles in cardiac hypertrophy (CH). However, the interaction between CaMKII and CnA-NFAT signaling remains unclear. H9c2 cells were cultured and treated with angiotensin II (Ang II) with or without silenced CaMKII (siCaMKII) and cyclosporine A (CsA, a calcineurin inhibitor) and subsequently treated with Wenxin Keli (WXKL). Patch clamp recording was conducted to assess L-type Ca current (I), and the expression of proteins involved in signaling pathways was measured by western blotting. Myocardial cytoskeletal protein and nuclear translocation of target proteins were assessed by immunofluorescence. The results indicated that siCaMKII suppressed Ang II-induced CH, as evidenced by reduced cell surface area and I. Notably, siCaMKII inhibited Ang II-induced activation of CnA and NFATc4 nuclear transfer. Inflammatory signaling was inhibited by siCaMKII and WXKL. Interestingly, CsA inhibited CnA-NFAT pathway expression but activated CaMKII signaling. In conclusion, siCaMKII may improve CH, possibly by blocking CnA-NFAT and MyD88 signaling, and WXKL has a similar effect. These data suggest that inhibiting CaMKII, but not CnA, may be a promising approach to attenuate CH and arrhythmia progression.
先前的研究表明，钙/钙调蛋白依赖性蛋白激酶II（CaMKII）和钙调神经磷酸酶A-活化T细胞核因子（CnA-NFAT）信号通路在心脏肥大（CH）中起关键作用。然而，CaMKII与CnA-NFAT信号之间的相互作用仍不清楚。培养H9c2细胞，用血管紧张素II（Ang II）处理，同时或不沉默CaMKII（siCaMKII）和环孢素A（CsA，一种钙调神经磷酸酶抑制剂），随后用温心颗粒（WXKL）处理。进行膜片钳记录以评估L型钙电流（I），并通过蛋白质印迹法测量信号通路中相关蛋白质的表达。通过免疫荧光评估心肌细胞骨架蛋白和靶蛋白的核转位。结果表明，siCaMKII抑制Ang II诱导的CH，细胞表面积和I降低证明了这一点。值得注意的是，siCaMKII抑制Ang II诱导的CnA激活和NFATc4核转位。siCaMKII和WXKL抑制炎症信号。有趣的是，CsA抑制CnA-NFAT通路表达，但激活CaMKII信号。总之，siCaMKII可能通过阻断CnA-NFAT和MyD88信号改善CH，WXKL有类似作用。这些数据表明，抑制CaMKII而非CnA可能是减轻CH和心律失常进展的一种有前景的方法。
