Dasatinib Attenuates Pressure Overload Induced Cardiac Fibrosis in a Murine Transverse Aortic Constriction Model.
Dasatinib Attenuates Pressure Overload Induced Cardiac Fibrosis in a Murine Transverse Aortic Constriction Model.
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DOI：
10.1371/journal.pone.0140273
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Kuppuswamy D
中科院分区：
文献类型：
--
作者：
Balasubramanian S;Pleasant DL;Kasiganesan H;Quinones L;Zhang Y;Sundararaj KP;Roche S;O'Connor R;Bradshaw AD;Kuppuswamy D
慢性压力超负荷（PO）引起的反应性心脏纤维化会损害心室功能并导致充血性心力衰竭。我们探讨非受体酪氨酸激酶 (NTK) 是否通过激活心脏成纤维细胞 (CFb) 在纤维化中发挥关键作用，并可能作为减少 PO 诱导的心脏纤维化的靶点。我们的研究是在横主动脉缩窄 (TAC) 诱导的 PO 小鼠心肌中进行的。通过腹膜内植入的微型渗透泵以 0.44 mg/kg/天施用酪氨酸激酶抑制剂达沙替尼，可减少 PO 诱导的细胞外基质 (ECM) 蛋白的积累，并改善左心室的几何形状和功能。此外，达沙替尼治疗抑制 NTK 激活（主要是 Pyk2 和 Fak）并降低 PO 心肌中 FSP1 阳性细胞的水平。使用培养小鼠 CFb 的体外研究表明，50 nM 的达沙替尼治疗可减少：(i) 胶原蛋白和纤连蛋白的细胞外积累，(ii) Pyk2 的基础激活和 PDGF 刺激的激活，(iii) Ki67、SKP2 和组蛋白-H2B 的核积累，以及 (iv) PDGF 刺激的 CFb 增殖和迁移。然而，达沙替尼不影响体内给药后心室组织中或体外处理后分离细胞中的心肌细胞形态。培养细胞中达沙替尼的质谱定量表明，CFb 对达沙替尼的摄取大于心肌细胞对达沙替尼的摄取。达沙替尼治疗主要抑制 CFb 和心肌细胞中的 PDGF，但不抑制胰岛素刺激的信号传导（Erk 与 Akt 激活）。这些数据表明，低于化疗剂量的达沙替尼治疗能够减少肥厚相关的纤维化并改善心室功能。
Reactive cardiac fibrosis resulting from chronic pressure overload (PO) compromises ventricular function and contributes to congestive heart failure. We explored whether nonreceptor tyrosine kinases (NTKs) play a key role in fibrosis by activating cardiac fibroblasts (CFb), and could potentially serve as a target to reduce PO-induced cardiac fibrosis. Our studies were carried out in PO mouse myocardium induced by transverse aortic constriction (TAC). Administration of a tyrosine kinase inhibitor, dasatinib, via an intraperitoneally implanted mini-osmotic pump at 0.44 mg/kg/day reduced PO-induced accumulation of extracellular matrix (ECM) proteins and improved left ventricular geometry and function. Furthermore, dasatinib treatment inhibited NTK activation (primarily Pyk2 and Fak) and reduced the level of FSP1 positive cells in the PO myocardium. In vitro studies using cultured mouse CFb showed that dasatinib treatment at 50 nM reduced: (i) extracellular accumulation of both collagen and fibronectin, (ii) both basal and PDGF-stimulated activation of Pyk2, (iii) nuclear accumulation of Ki67, SKP2 and histone-H2B and (iv) PDGF-stimulated CFb proliferation and migration. However, dasatinib did not affect cardiomyocyte morphologies in either the ventricular tissue after in vivo administration or in isolated cells after in vitro treatment. Mass spectrometric quantification of dasatinib in cultured cells indicated that the uptake of dasatinib by CFb was greater that that taken up by cardiomyocytes. Dasatinib treatment primarily suppressed PDGF but not insulin-stimulated signaling (Erk versus Akt activation) in both CFb and cardiomyocytes. These data indicate that dasatinib treatment at lower doses than that used in chemotherapy has the capacity to reduce hypertrophy-associated fibrosis and improve ventricular function.