S-nitrosylation of c-Jun N-terminal kinase mediates pressure overload-induced cardiac dysfunction and fibrosis
S-nitrosylation of c-Jun N-terminal kinase mediates pressure overload-induced cardiac dysfunction and fibrosis
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c-Jun N 末端激酶的 S-亚硝基化介导压力超负荷引起的心脏功能障碍和纤维化
DOI：
10.1038/s41401-021-00674-9
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发表时间：
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影响因子：
8.2
通讯作者：
Ji Yong
中科院分区：
文献类型：
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作者：
Zhou Miao;Chen Ji-Yu;Chao Meng-Lin;Zhang Chao;Shi Zhi-Guang;Zhou Xue-Chun;Xie Li-Ping;Sun Shi-Xiu;Huang Zheng-Rong;Luo Shan-Shan;Ji Yong
心脏纤维化（CF）是一种不可逆的病理过程，几乎发生在所有心血管疾病中。c-Jun N-末端激酶（JNK）的磷酸化依赖性激活诱导心脏纤维化。然而，JNK的S-亚硝基化是否介导心脏纤维化仍然是一个悬而未决的问题。生物素开关试验证实，S-亚硝基化的JNK（SNO-JNK）显着增加的心脏组织中的肥大患者，横主动脉缩窄（TAC）小鼠，自发性高血压大鼠（SHR），和新生大鼠心脏成纤维细胞（NRCFs）血管紧张素II（Ang II）刺激。应用JNK中丙氨酸对半胱氨酸的位点间取代来确定S-亚硝基化位点。在Cys 116和Cys 163处均发生S-亚硝基化，并且丙氨酸取代半胱氨酸116和半胱氨酸163（C116/163 A）抑制Ang II诱导的肌成纤维细胞转化。我们进一步证实了S-亚硝基化的来源是诱导型一氧化氮合酶（iNOS）。iNOS抑制剂1400 W可阻断Ang Ⅱ对NRCF的促纤维化作用。通过染色质免疫沉淀和EMSA测定，SNO-JNK促进JNK的核转位，增加c-Jun的磷酸化，并诱导AP-1的转录活性。最后，WT和iNOS-/-小鼠接受TAC和iNOS敲除，减少了SNO-JNK，减轻了心脏纤维化。我们的研究结果证明了iNOS诱导的SNO-JNK增加JNK通路活性并加速心脏纤维化的另一种机制。以SNO-JNK为靶点可能成为抗心肌纤维化的一种新的治疗策略。
Cardiac fibrosis (CF) is an irreversible pathological process that occurs in almost all kinds of cardiovascular diseases. Phosphorylation-dependent activation of c-Jun N-terminal kinase (JNK) induces cardiac fibrosis. However, whether S-nitrosylation of JNK mediates cardiac fibrosis remains an open question. A biotin-switch assay confirmed that S-nitrosylation of JNK (SNO-JNK) increased significantly in the heart tissues of hypertrophic patients, transverse aortic constriction (TAC) mice, spontaneously hypertensive rats (SHRs), and neonatal rat cardiac fibroblasts (NRCFs) stimulated with angiotensin II (Ang II). Site to site substitution of alanine for cysteine in JNK was applied to determine the S-nitrosylated site. S-Nitrosylation occurred at both Cys116 and Cys163 and substitution of alanine for cysteine 116 and cysteine 163 (C116/163A) inhibited Ang II-induced myofibroblast transformation. We further confirmed that the source of S-nitrosylation was inducible nitric oxide synthase (iNOS). 1400 W, an inhibitor of iNOS, abrogated the profibrotic effects of Ang II in NRCFs. Mechanistically, SNO-JNK facilitated the nuclear translocation of JNK, increased the phosphorylation of c-Jun, and induced the transcriptional activity of AP-1 as determined by chromatin immunoprecipitation and EMSA. Finally, WT and iNOS−/−mice were subjected to TAC and iNOS knockout reduced SNO-JNK and alleviated cardiac fibrosis. Our findings demonstrate an alternative mechanism by which iNOS-induced SNO-JNK increases JNK pathway activity and accelerates cardiac fibrosis. Targeting SNO-JNK might be a novel therapeutic strategy against cardiac fibrosis.