Objective:To investigate whether cardiac mast cells(MCs)participate in pressure overload-induced myocardial hypertrophy through the regulation of transient receptor potential vanilloid 4(TRPV4).Methods:Pressure overlo...Objective:To investigate whether cardiac mast cells(MCs)participate in pressure overload-induced myocardial hypertrophy through the regulation of transient receptor potential vanilloid 4(TRPV4).Methods:Pressure overload-induced myocardial hypertrophy was induced via abdominal aortic constriction(AAC).Myocardial hypertrophy was evaluated by measuring the heart weight index(HW/BW),lung weight index(LW/BW),ratio of heart weight to tibia length(HW/TL),ratio of lung weight to tibia length(LW/TL),and cross-sectional area of myocardial cells.qRT-PCR was used to detect the mRNA expression of TRPV4.Western blotting was used to detect the protein expression of TRPV4,mast cell tryptase,myosin heavy chain beta(β-MHC),calcineurin A(CnA),and nuclear factor of activated T-cell c3(NFATc3).ELISA was used to measure the levels of brain natriuretic peptide(BNP)and histamine.Fluo4 AM was used to detect the calcium signal in H9c2 myocardial cells.Results:Compared with those of the sham rats,the myocardial mast cells,tryptase,HW/BW,LW/BW,HW/TL,and LW/TL,the cross-sectional area of the myocardial cells,and the expression ofβ-MHC,TRPV4,CnA,and NFATc3 in the myocardial tissue and the serum BNP of the AAC-treated rats increased significantly,whereas the MC stabilizer cromolyn sodium(CS)reversed these indicators.In H9c2 cardiomyocytes,treatment with histamine and the TRPV4 agonist GSK1016790A upregulated the expression of TRPV4,β-MHC,BNP,CnA and NFATc3 and increased calcium ion influx,whereas these effects were inhibited by the H2 receptor inhibitor famotidine and the TRPV4 inhibitor HC067047.Conclusion:Cardiac MCs participate in pressure overload-induced myocardial hypertrophy through the upregulation of TRPV4 via its mediator histamine,and the Ca^(2+)/CnA/NFATc3 signaling pathway is involved in this process.展开更多
活化T细胞核因子3(Nuclear factor of activated T cells 3,NFATc3)是NFAT家族的重要成员,在肌纤维类型转换中具有重要的作用。为探讨NFATc3 mRNA在不同品种鸡不同类型肌肉中的表达情况,以生长速度具有较大差异的清远麻鸡和隐性白羽鸡...活化T细胞核因子3(Nuclear factor of activated T cells 3,NFATc3)是NFAT家族的重要成员,在肌纤维类型转换中具有重要的作用。为探讨NFATc3 mRNA在不同品种鸡不同类型肌肉中的表达情况,以生长速度具有较大差异的清远麻鸡和隐性白羽鸡为材料,采用实时荧光定量PCR方法,检测不同发育时期趾长伸肌和比目鱼肌中NFATc3 mRNA的表达情况。结果发现,NFATc3 mRNA在两个品种骨骼肌的变化趋势基本一致,1日龄时,NFATc3在趾长伸肌和比目鱼肌中的表达量都较高,7日龄时显著下降,21日龄时出现上升,之后表达量趋于平稳;品种间比较,比目鱼肌中在21、49日龄时,清远麻鸡的基因表达量显著高于隐性白羽鸡(P<0.05),趾长伸肌中则在1、21日龄时,清远麻鸡的基因表达量显著高于隐性白羽鸡(P<0.05);同一品种不同类型肌肉间比较,总体上都是趾长伸肌的表达量高于比目鱼肌。提示,鸡骨骼肌中NFATc3 mRNA表达变化规律不存在品种差异,NFATc3可能在鸡不同类型肌肉肌纤维的发育中起重要作用。展开更多
ABSTRACT The molecular signaling pathway linked to hyper-trophy of the anti-gravity/postural soleus muscle af-ter mechanical overloading has not been identified. Using Western blot and immunohistochemical analy-ses, w...ABSTRACT The molecular signaling pathway linked to hyper-trophy of the anti-gravity/postural soleus muscle af-ter mechanical overloading has not been identified. Using Western blot and immunohistochemical analy-ses, we investigated whether the amounts of NFATc3, GSK-3?, NFATc1, and neonatal MHC change in the mechanically overloaded soleus muscle after cyc-losporine A (CsA) treatment. Adult male ICR mice were subjected to a surgical ablation of the gas-trocnemius muscle and treated with either CsA (25 mg/Kg) or vehicle once daily. They were sacrificed at 2, 4, 7, 10, and 14 days post-injury. Mechanical over-loading resulted in a significant increase in the wet weight and the cross-sectional area of slow and fast fibers of the soleus muscle in placebo-treated mice but not CsA-treated mice. After 4 days of mechanical overloading, we observed a similar co-localization of neonatal MHC and NFATc3 in several myotubes of both mice. The placebo-treated mice possessed larger myotubes with neonatal MHC than CsA-treated mice. At 7 days, mechanical overloading induced marked expression of neonatal MHC in myotubes and/or myofibers. Such neonatal MHC-positive fibers emerged less often in the hypertrophied soleus mus-cle subjected to treatment with CsA. CsA treatment did not significantly change the amount of GSK-3? protein in the soleus muscle. The modulation of growth in neonatal MHC-positive myofibers by CsA treatment may inhibit the hypertrophic process in the soleus muscle after mechanical overloading.展开更多
This study investigated the role of the nuclear factor of activated T cells c3(NFATc3)in vascular smooth muscle cells(VSMCs)during aortic aneurysm and dissection(AAD)progression and the underlying molecular mechanisms...This study investigated the role of the nuclear factor of activated T cells c3(NFATc3)in vascular smooth muscle cells(VSMCs)during aortic aneurysm and dissection(AAD)progression and the underlying molecular mechanisms.Cytoplasmic and nuclear NFATc3 levels were elevated in human and mouse AAD.VSMC–NFATc3 deletion reduced thoracic AAD(TAAD)and abdominal aortic aneurysm(AAA)progression in mice,contrary to VSMC–NFATc3 overexpression.VSMC–NFATc3 deletion reduced extracellular matrix(ECM)degradation and maintained the VSMC contractile phenotype.Nuclear NFATc3 targeted and transcriptionally upregulated matrix metalloproteinase 9(MMP9)and MMP2,promoting ECM degradation and AAD development.NFATc3 promoted VSMC phenotypic switching by binding to eukaryotic elongation factor 2(eEF2)and inhibiting its phosphorylation in the VSMC cytoplasm.Restoring eEF2 reversed the beneficial effects in VSMC-specific NFATc3-knockout mice.Cabamiquine—targets eEF2 and inhibits protein synthesis—inhibited AAD development and progression in VSMC-NFATc3-overexpressing mice.VSMC–NFATc3 promoted VSMC switch and ECM degradation while exacerbating AAD development,making it a novel potential therapeutic target for preventing and treating AAD.展开更多
基金supported by grants from the National Natural Science Foundation of China(No.30872716)the Natural Science Foundation of Hubei Province(No.2015CFB288).
文摘Objective:To investigate whether cardiac mast cells(MCs)participate in pressure overload-induced myocardial hypertrophy through the regulation of transient receptor potential vanilloid 4(TRPV4).Methods:Pressure overload-induced myocardial hypertrophy was induced via abdominal aortic constriction(AAC).Myocardial hypertrophy was evaluated by measuring the heart weight index(HW/BW),lung weight index(LW/BW),ratio of heart weight to tibia length(HW/TL),ratio of lung weight to tibia length(LW/TL),and cross-sectional area of myocardial cells.qRT-PCR was used to detect the mRNA expression of TRPV4.Western blotting was used to detect the protein expression of TRPV4,mast cell tryptase,myosin heavy chain beta(β-MHC),calcineurin A(CnA),and nuclear factor of activated T-cell c3(NFATc3).ELISA was used to measure the levels of brain natriuretic peptide(BNP)and histamine.Fluo4 AM was used to detect the calcium signal in H9c2 myocardial cells.Results:Compared with those of the sham rats,the myocardial mast cells,tryptase,HW/BW,LW/BW,HW/TL,and LW/TL,the cross-sectional area of the myocardial cells,and the expression ofβ-MHC,TRPV4,CnA,and NFATc3 in the myocardial tissue and the serum BNP of the AAC-treated rats increased significantly,whereas the MC stabilizer cromolyn sodium(CS)reversed these indicators.In H9c2 cardiomyocytes,treatment with histamine and the TRPV4 agonist GSK1016790A upregulated the expression of TRPV4,β-MHC,BNP,CnA and NFATc3 and increased calcium ion influx,whereas these effects were inhibited by the H2 receptor inhibitor famotidine and the TRPV4 inhibitor HC067047.Conclusion:Cardiac MCs participate in pressure overload-induced myocardial hypertrophy through the upregulation of TRPV4 via its mediator histamine,and the Ca^(2+)/CnA/NFATc3 signaling pathway is involved in this process.
文摘ABSTRACT The molecular signaling pathway linked to hyper-trophy of the anti-gravity/postural soleus muscle af-ter mechanical overloading has not been identified. Using Western blot and immunohistochemical analy-ses, we investigated whether the amounts of NFATc3, GSK-3?, NFATc1, and neonatal MHC change in the mechanically overloaded soleus muscle after cyc-losporine A (CsA) treatment. Adult male ICR mice were subjected to a surgical ablation of the gas-trocnemius muscle and treated with either CsA (25 mg/Kg) or vehicle once daily. They were sacrificed at 2, 4, 7, 10, and 14 days post-injury. Mechanical over-loading resulted in a significant increase in the wet weight and the cross-sectional area of slow and fast fibers of the soleus muscle in placebo-treated mice but not CsA-treated mice. After 4 days of mechanical overloading, we observed a similar co-localization of neonatal MHC and NFATc3 in several myotubes of both mice. The placebo-treated mice possessed larger myotubes with neonatal MHC than CsA-treated mice. At 7 days, mechanical overloading induced marked expression of neonatal MHC in myotubes and/or myofibers. Such neonatal MHC-positive fibers emerged less often in the hypertrophied soleus mus-cle subjected to treatment with CsA. CsA treatment did not significantly change the amount of GSK-3? protein in the soleus muscle. The modulation of growth in neonatal MHC-positive myofibers by CsA treatment may inhibit the hypertrophic process in the soleus muscle after mechanical overloading.
基金supported by grants from the National Natural Science Foundation of China(82470476,82200447)to Xiu Liu(82470418,82170274)to Shaoyi Zheng+8 种基金(82001230)to Yonghua Tuo(82070312)to Sijia LiangGuangdong Provincial International Science and Technology Cooperation Project(2022A0505050036,China)to Shaoyi ZhengGuangzhou Science and Technology Projects&Key Research Development Program(202206010065,China)to Shaoyi ZhengGuangdong Basic and Applied Basic Research Foundation(2022A1515011747,China)to Shaoyi Zheng(2024A1515013074)to Xiu LiuScience and Technology Projects in Guangzhou(2024A04J5091,China)to Xiu Liu(2024A03J0196)to Yonghua TuoOutstanding Youths Development Scheme of Nanfang Hospital of Southern Medical University(2023J004,China)to Xiu Liu.
文摘This study investigated the role of the nuclear factor of activated T cells c3(NFATc3)in vascular smooth muscle cells(VSMCs)during aortic aneurysm and dissection(AAD)progression and the underlying molecular mechanisms.Cytoplasmic and nuclear NFATc3 levels were elevated in human and mouse AAD.VSMC–NFATc3 deletion reduced thoracic AAD(TAAD)and abdominal aortic aneurysm(AAA)progression in mice,contrary to VSMC–NFATc3 overexpression.VSMC–NFATc3 deletion reduced extracellular matrix(ECM)degradation and maintained the VSMC contractile phenotype.Nuclear NFATc3 targeted and transcriptionally upregulated matrix metalloproteinase 9(MMP9)and MMP2,promoting ECM degradation and AAD development.NFATc3 promoted VSMC phenotypic switching by binding to eukaryotic elongation factor 2(eEF2)and inhibiting its phosphorylation in the VSMC cytoplasm.Restoring eEF2 reversed the beneficial effects in VSMC-specific NFATc3-knockout mice.Cabamiquine—targets eEF2 and inhibits protein synthesis—inhibited AAD development and progression in VSMC-NFATc3-overexpressing mice.VSMC–NFATc3 promoted VSMC switch and ECM degradation while exacerbating AAD development,making it a novel potential therapeutic target for preventing and treating AAD.
