卵泡抑素样蛋白4通过活化T细胞核因子抑制病理性心肌肥厚的发生发展
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摘要
目的通过应用卵泡抑素样蛋白4(FSTL4)基因敲除小鼠和过表达FSTL4的细胞从体内和体外两方面研究FSTL4在病理性心肌肥厚中的功能和作用机制。方法对8只野生型小鼠和8只FSTL4基因敲除小鼠进行主动脉弓缩窄手术,构建压力超负荷诱导的心肌肥厚模型。术后4周行超声检测评估心脏的结构与功能,并通过组织病理学染色和实时荧光定量聚合酶链式反应评价心肌肥厚和纤维化的程度。在H9C2细胞中过表达FSTL4并以血管紧张素Ⅱ(AngⅡ)刺激,通过免疫荧光染色、实时荧光定量聚合酶链式反应和双荧光素酶报告基因检测,探究体外条件下FSTL4对心肌细胞肥大的作用。结果主动脉弓缩窄手术后4周,与野生型小鼠相比,FSTL4基因敲除小鼠的心肌肥厚和心肌纤维化程度加重,心脏功能减退。在H9C2细胞中过表达FSTL4抑制了AngⅡ诱导的心肌细胞肥大。进一步探索发现FSTL4抑制活化T细胞核因子(NFAT)的转录活性。结论 FSTL4基因缺失促进病理性心肌肥厚、心脏纤维化和心功能的恶化,过表达FSTL4能够抑制NFAT的转录活性,抑制心肌细胞肥大。
Objective To investigate the function and mechanism of Follistatin-like 4(FSTL4) in pathological cardiac hypertrophy in vivo and in vitro using FSTL4 knockout mice and overexpressing cells. Methods A transverse aortic constriction-induced cardiac hypertrophy model was established in eight FSTL4 knockout mice in parallel with wild-type control mice.Four weeks after the operation, cardiac structure and function was assessed by echocardiography measurement. Histological staining and real-time quantitative PCR were performed to detect the degree of cardiac hypertrophy and fibrosis.To evaluate the role of FSTL4 in cardiomyocyte enlargement, FSTL4 was overexpressed in H9 C2 cells that were then stimulated with angiotensin Ⅱ(AngⅡ).Immunofluorescence staining,real-time quantitative PCR and luciferase were performed to investigate the effect of FSTL4 on cardiac hypertrophy in vitro. Results Four weeks after transverse aortic constriction, FSTL4 knockout mice showed more severe cardiac hypertrophy, myocardial fibrosis and cardiac dysfunction than wild-type mice. Overexpression of FSTL4 in H9 C2 cells protected against AngⅡ-induced cardiomyocyte hypertrophy in vitro.Further exploration revealed that FSTL4 inhibited the transcriptional activity of nuclear factor of activated T cells. Conclusion FSTL4 deletion promotes pathological cardiac hypertrophy, myocardial fibrosis and cardiac dysfunction. Overexpression of FSTL4 can inhibit NFAT transcriptional activity and cardiomyocyte hypertrophy.
Objective To investigate the function and mechanism of Follistatin-like 4(FSTL4) in pathological cardiac hypertrophy in vivo and in vitro using FSTL4 knockout mice and overexpressing cells. Methods A transverse aortic constriction-induced cardiac hypertrophy model was established in eight FSTL4 knockout mice in parallel with wild-type control mice.Four weeks after the operation, cardiac structure and function was assessed by echocardiography measurement. Histological staining and real-time quantitative PCR were performed to detect the degree of cardiac hypertrophy and fibrosis.To evaluate the role of FSTL4 in cardiomyocyte enlargement, FSTL4 was overexpressed in H9 C2 cells that were then stimulated with angiotensin Ⅱ(AngⅡ).Immunofluorescence staining,real-time quantitative PCR and luciferase were performed to investigate the effect of FSTL4 on cardiac hypertrophy in vitro. Results Four weeks after transverse aortic constriction, FSTL4 knockout mice showed more severe cardiac hypertrophy, myocardial fibrosis and cardiac dysfunction than wild-type mice. Overexpression of FSTL4 in H9 C2 cells protected against AngⅡ-induced cardiomyocyte hypertrophy in vitro.Further exploration revealed that FSTL4 inhibited the transcriptional activity of nuclear factor of activated T cells. Conclusion FSTL4 deletion promotes pathological cardiac hypertrophy, myocardial fibrosis and cardiac dysfunction. Overexpression of FSTL4 can inhibit NFAT transcriptional activity and cardiomyocyte hypertrophy.
引文
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2 Kannan A,Janardhanan R.Hypertension as a risk factor for heart failure[J].Current Hypertension Reports,2014,16(7)
3 Fu S,Li Y,Wu Y,et al.Icariside Ⅱ attenuates myocardial fibrosis by inhibiting nuclear factor-κB and the TGF-β1/Smad2 signalling pathway in spontaneously hypertensive rats[J].Biomedicine & Pharmacotherapy,2018,100:64
4 Liu L,He X,Zhao M,et al.Regulation of DNA methylation and 2-OG/TET signaling by choline alleviated cardiac hypertrophy in spontaneously hypertensive rats[J].Journal of Molecular and Cellular Cardiology,2019,128:26
5 Tsuchida K,Arai K Y,Kuramoto Y,et al.Identification and characterization of a novel follistatin-like protein as a binding protein for the TGF-β family[J].Journal of Biological Chemistry,2000,275(52):40 788
6 Guo Y,Tomlinson B,Chu T,et al.A genome-wide linkage and association scan reveals novel loci for hypertension and blood pressure traits[J].PLoS One,2012,7(2):e31489
7 Luke MM,O'Meara ES,Rowland CM,et al.Gene variants associated with ischemic stroke[J].Stroke,2009,40(2):363
8 Bian Z,Huang H,Jiang H,et al.LIM and cysteine-rich domains 1 regulates cardiac hypertrophy by targeting calcineurin/nuclear factor of activated T cells signaling[J].Hypertension,2010,55(2):257
9 Li H,He C,Feng J,et al.Regulator of G protein signaling 5 protects against cardiac hypertrophy and fibrosis during biomechanical stress of pressure overload[J].Proceedings of the National Academy of Sciences,2010,107(31):13 818
10 Molkentin J.Calcineurin-NFAT signaling regulates the cardiac hypertrophic response in coordination with the MAPKs[J].Cardiovascular Research,2004,63(3):467
11 Gao L,Yao R,Liu Y,et al.Isorhamnetin protects against cardiac hypertrophy through blocking PI3K–AKT pathway[J].Molecular and Cellular Biochemistry,2017,429(1-2):167
12 Heineke J,Molkentin JD.Regulation of cardiac hypertrophy by intracellular signalling pathways[J].Nature Reviews Molecular Cell Biology,2006,7(8):589
13 Tham YK,Bernardo BC,Ooi JYY,et al.Pathophysiology of cardiac hypertrophy and heart failure:signaling pathways and novel therapeutic targets[J].Archives of Toxicology,2015,89(9):1 401
14 Yonehara K,Shintani T,Suzuki R,et al.Expression of SPIG1 reveals development of a retinal ganglion cell subtype projecting to the medial terminal nucleus in the mouse[J].PloS one,2008,3(2):e1 533
15 Suzuki R,Matsumoto M,Fujikawa A,et al.SPIG1 negatively regulates BDNF maturation[J].Journal of Neuroscience,2014,34(9):3 429
16 Li X,Lan Y,Wang Y,et al.Telmisartan suppresses cardiac hypertrophy by inhibiting cardiomyocyte apoptosis via the NFAT/ANP/BNP signaling pathway[J].Mol Med Rep,2017,15(5):2 574