Apurine/pyrimidine-free endonuclease 1(APEX1)is a multifunctional enzyme that contributes to oxidization-mediated DNA-cleaved base excision repair and redox activation of transcription factors.However,the role of APEX...Apurine/pyrimidine-free endonuclease 1(APEX1)is a multifunctional enzyme that contributes to oxidization-mediated DNA-cleaved base excision repair and redox activation of transcription factors.However,the role of APEX1 during cardiomyocyte oxidative stress injury is not completely understood.In the present study,whether APEX1 protects oxidative damage-induced cardiomyocytes was investigated.mRNA and protein expression levels of APEX1 were downregulated in the mouse model of cardiac ischemia-reperfusion injury.Furthermore,the expression of APEX1 in hydrogen peroxide(H 2 O 2)-treated neonatal mice cardiomyocytes was also decreased.APEX1 knockdown aggravated H 2 O 2-treated cardiomyocyte apoptosis indexes.By contrast,APEX1 overexpression reversed H 2 O 2-induced oxidative damage,as demonstrated by decreased caspase 3 and Bax expression levels.Moreover,homeobox A5 upregulated APEX1.The results of the present study indicated that APEX1 displayed protective effects against oxidative damage,suggesting that APEX1 may serve as a unique protective strategy for cardiac ischemia-reperfusion injury.展开更多
MicroRNAs(miRNAs) are endogenous 20 -23 -nucleotide (nt) -containing small non-coding RNAs that negatively regulate gene expression in diverse biological and pathological processes,including cell differentiation,proli...MicroRNAs(miRNAs) are endogenous 20 -23 -nucleotide (nt) -containing small non-coding RNAs that negatively regulate gene expression in diverse biological and pathological processes,including cell differentiation,proliferation, apoptosis,heart disease and human cancers.We investigated miR-133 expression and its potential role in a high glucose-induced myocardium in Streptozotocin(STZ)-induced C57bl6 mouse model of diabetes.miR-133 expression was significantly increased in myocardium in a time-dependent manner after STZ treatment.IGF1 receptor(IGF1R) protein was dramatically decreased without obvious up-regulation of its mRNA level post hyperglycemia.IGF1R protein level was decreaed with increase of its transcript level in neonatal mouse ventricular cardiomyocytes induced by high D-glucose concentration. Dual luciferase assay revealed that miR133 could interact with specific sites in the 3’UTR of IGF1R gene.p-ERK and p-Akt levels were reduced in neonatal mouse cardiomyocytes over-expressed with miR133 after IGF treatment.Introduction of functional miR-133,IGF1R siRNA into neonatal mouse cardiomyocytes could enhance cardiomyocyte apoptosis.These results implicate that miR-133 is involved in contributing to high glucose-induced cardiomyocyte apoptosis via regulating IGF1R expression post-transcriptionally.展开更多
Activation of the local renin-angiotensin system(RAS)promotes cardiomyocyte apoptosis and cardiac remodeling after acute myocardial infarction(AMI).As an anti-RAS drug,the effect of Valsartan in the early stage of acu...Activation of the local renin-angiotensin system(RAS)promotes cardiomyocyte apoptosis and cardiac remodeling after acute myocardial infarction(AMI).As an anti-RAS drug,the effect of Valsartan in the early stage of acute MI is limited by its low drug concentration in the heart and low dosage.Here,by exploiting the inherent nature of neutrophils migrating to the injured myocardium and the local low-pH microenvironment caused by ischemia and hypoxia after myocardial infarction,we designed nanocarrier(NSLP)-hybridized neutrophil membranes and pH-sensitive liposomes(SLPs)for the delivery of Valsartan(NSLPVal).These functional nanocarriers could mimic neutrophils and are homed to the injured heart;they were also found to respond to a low-pH microenvironment.In the mouse model of MI,we found that NSLP-Val could target the infarct marginal zone and release Valsartan locally in the low-pH microenvironment without affecting hemodynamic stability.Further,locally released angiotensin receptor inhibitors reduced the infarct size and inflammatory response by inhibiting cardiomyocytes.Ultimately,NSLP-Val improved cardiac function and inhibited cardiac hypertrophy and fibrosis.展开更多
BACKGROUND Heart failure(HF)is a global health problem characterized by impaired heart function.Cardiac remodeling and cell death contribute to the development of HF.Although treatments such as digoxin and angiotensin...BACKGROUND Heart failure(HF)is a global health problem characterized by impaired heart function.Cardiac remodeling and cell death contribute to the development of HF.Although treatments such as digoxin and angiotensin receptor blocker drugs have been used,their effectiveness in reducing mortality is uncertain.Researchers are exploring the use of adipose-derived mesenchymal stem cell(ADMSC)exosomes(Exos)as a potential therapy for HF.These vesicles,secreted by cells,may aid in tissue repair and regulation of inflammation and immune responses.However,further investigation is needed to understand the specific role of these vesicles in HF treatment.AIM To investigate the mechanism of extracellular vesicles produced by ADMSC s in the treatment of HF.METHODS Exogenous surface markers of ADMSCs were found,and ADMSCs were cultured.RESULTS The identification of surface markers showed that the surface markers CD44 and CD29 of adipose-derived stem cells(ADSCs)were well expressed,while the surface markers CD45 and CD34 of ADSCs were negative,so the cultured cells were considered ADSCs.Western blotting detected the Exo surface marker protein,which expressed CD63 protein but did not express calnexin protein,indicating that ADSC-derived Exos were successfully extracted.CONCLUSION The secretion of MSCs from adipose tissue can increase ATP levels,block cardiomyocyte apoptosis,and enhance the heart function of animals susceptible to HF.The inhibition of Bax,caspase-3 and p53 protein expression may be related to this process.展开更多
Clinical application of doxorubicin(DOX)is heavily hindered by DOX cardiotoxicity.Several theories were postulated for DOX cardiotoxicity including DNA damage and DNA damage response(DDR),although the mechanism(s)invo...Clinical application of doxorubicin(DOX)is heavily hindered by DOX cardiotoxicity.Several theories were postulated for DOX cardiotoxicity including DNA damage and DNA damage response(DDR),although the mechanism(s)involved remains to be elucidated.This study evaluated the potential role of TBC domain family member 15(TBC1D15)in DOX cardiotoxicity.Tamoxifen-induced cardiac-specific Tbcldi5 knockout(Tbcldi5^(CKO))or Tbcldi5 knockin(Tbcldi5^(CKI))male mice were challenged with a single dose of DOx prior to cardiac assessment 1 week or 4 weeks following DOX challenge.Adenoviruses encoding TBC1D15 or containing shRNA targeting Tbcld15 were used for Tbcld15 overexpression or knockdown in isolated primary mouse cardiomyocytes.Our results re-vealed that DOX evoked upregulation of TBC1D15 with compromised myocardial function and overt mortality,the effects of which were ameliorated and accentuated by Tbcldi5 deletion and Tbcld15 overexpression,respectively.DOX overtly evoked apoptotic cell death,the effect of which was alleviated and exacerbated by Tbcld15 knockout and overexpression,respectively.Meanwhile,DOX provoked mitochondrial membrane potential collapse,oxidative stress and DNA damage,the effects of which were mitigated and exacerbated by Tbcld15 knockdown and overexpression,respectively.Further scrutiny revealed that TBC1D15 fostered cytosolic accumulation of the cardinal DDR element DNA-dependent protein kinase catalytic subunit(DNA-PKcs).Liquid chromatography-tandem mass spectrometry and coimmunoprecipitation denoted an interaction between TBCID15 and DNA-PKcs at the segment 594-624 of TBC1D15.Moreover,overexpression of TBC1D15 mutant(A594-624,deletion of segment 594-624)failed to elicit accentuation of DOX-induced cytosolic retention of DNA-PKcs,DNA damage and cardiomyocyte apoptosis by TBC1D15 wild type.However,Tbcld15 deletion ameliorated DOXinduced cardiomyocyte contractile anomalies,apoptosis,mitochondrial anomalies,DNA damage and cytosolic DNA-PKcs accumulation,which were canceled off by DNA-PKcs inhibition or ATM activation.Taken together,our findings denoted a pivotal role for TBCID15 in DOX-induced DNA damage,mitochondrial injury,and apoptosis possibly through binding with DNA-PKcs and thus gate-keeping its cytosolic retention,a route to accentuation of cardiac contractile dysfunction in DOX-induced cardiotoxicity.展开更多
As an effective anticancer drug, the clinical limitation of doxorubicin(Dox) is the time-and dose-dependent cardiotoxicity. Yes-associated protein 1(YAP1) interacts with transcription factor TEA domain 1(TEAD1) and pl...As an effective anticancer drug, the clinical limitation of doxorubicin(Dox) is the time-and dose-dependent cardiotoxicity. Yes-associated protein 1(YAP1) interacts with transcription factor TEA domain 1(TEAD1) and plays an important role in cell proliferation and survival. However, the role of YAP1 in Dox-induced cardiomyopathy has not been reported. In this study, the expression of YAP1 was reduced in clinical human failing hearts with dilated cardiomyopathy and Dox-induced in vivo and in vitro cardiotoxic model. Ectopic expression of Yap1 significantly blocked Dox-induced cardiomyocytes apoptosis in TEAD1 dependent manner. Isorhapontigenin(Isor) is a new derivative of stilbene and responsible for a wide range of biological processes. Here, we found that Isor effectively relieved Doxinduced cardiomyocytes apoptosis in a dose-dependent manner in vitro. Administration with Isor(30 mg/kg/day, intraperitoneally, 3 weeks) significantly protected against Dox-induced cardiotoxicity in mice. Interestingly, Isor increased Dox-caused repression in YAP1 and the expression of its target genes in vivo and in vitro. Knockout or inhibition of Yap1 blocked the protective effects of Isor on Dox-induced cardiotoxicity. In conclusion, YAP1 may be a novel target for Dox-induced cardiotoxicity and Isor might be a new compound to fight against Dox-induced cardiotoxicity by increasing YAP1 expression.展开更多
基金supported by the National Natural Science Foundation of China(Grant No.81900245 and 81770395).
文摘Apurine/pyrimidine-free endonuclease 1(APEX1)is a multifunctional enzyme that contributes to oxidization-mediated DNA-cleaved base excision repair and redox activation of transcription factors.However,the role of APEX1 during cardiomyocyte oxidative stress injury is not completely understood.In the present study,whether APEX1 protects oxidative damage-induced cardiomyocytes was investigated.mRNA and protein expression levels of APEX1 were downregulated in the mouse model of cardiac ischemia-reperfusion injury.Furthermore,the expression of APEX1 in hydrogen peroxide(H 2 O 2)-treated neonatal mice cardiomyocytes was also decreased.APEX1 knockdown aggravated H 2 O 2-treated cardiomyocyte apoptosis indexes.By contrast,APEX1 overexpression reversed H 2 O 2-induced oxidative damage,as demonstrated by decreased caspase 3 and Bax expression levels.Moreover,homeobox A5 upregulated APEX1.The results of the present study indicated that APEX1 displayed protective effects against oxidative damage,suggesting that APEX1 may serve as a unique protective strategy for cardiac ischemia-reperfusion injury.
文摘MicroRNAs(miRNAs) are endogenous 20 -23 -nucleotide (nt) -containing small non-coding RNAs that negatively regulate gene expression in diverse biological and pathological processes,including cell differentiation,proliferation, apoptosis,heart disease and human cancers.We investigated miR-133 expression and its potential role in a high glucose-induced myocardium in Streptozotocin(STZ)-induced C57bl6 mouse model of diabetes.miR-133 expression was significantly increased in myocardium in a time-dependent manner after STZ treatment.IGF1 receptor(IGF1R) protein was dramatically decreased without obvious up-regulation of its mRNA level post hyperglycemia.IGF1R protein level was decreaed with increase of its transcript level in neonatal mouse ventricular cardiomyocytes induced by high D-glucose concentration. Dual luciferase assay revealed that miR133 could interact with specific sites in the 3’UTR of IGF1R gene.p-ERK and p-Akt levels were reduced in neonatal mouse cardiomyocytes over-expressed with miR133 after IGF treatment.Introduction of functional miR-133,IGF1R siRNA into neonatal mouse cardiomyocytes could enhance cardiomyocyte apoptosis.These results implicate that miR-133 is involved in contributing to high glucose-induced cardiomyocyte apoptosis via regulating IGF1R expression post-transcriptionally.
基金The authors thank the Shiyanjia Lab(www.shiyanjia.com)for TEM measurements.The authors are grateful to Ms.Xiao Guo at the Joint Live Small Animal Imaging Laboratory of Fudan University Shanghai Medical College-PerkinElmer Company,for her technical support with the use of the in vivo imaging system.This work was financially supported by the National Key Research and Development Program of China(No.2016YFC1301200)the National Natural Science Foundation of China(Nos.82070281,81870269,and 82170524)Shanghai Clinical Research Center for Interventional Medicine(No.19MC1910300).
文摘Activation of the local renin-angiotensin system(RAS)promotes cardiomyocyte apoptosis and cardiac remodeling after acute myocardial infarction(AMI).As an anti-RAS drug,the effect of Valsartan in the early stage of acute MI is limited by its low drug concentration in the heart and low dosage.Here,by exploiting the inherent nature of neutrophils migrating to the injured myocardium and the local low-pH microenvironment caused by ischemia and hypoxia after myocardial infarction,we designed nanocarrier(NSLP)-hybridized neutrophil membranes and pH-sensitive liposomes(SLPs)for the delivery of Valsartan(NSLPVal).These functional nanocarriers could mimic neutrophils and are homed to the injured heart;they were also found to respond to a low-pH microenvironment.In the mouse model of MI,we found that NSLP-Val could target the infarct marginal zone and release Valsartan locally in the low-pH microenvironment without affecting hemodynamic stability.Further,locally released angiotensin receptor inhibitors reduced the infarct size and inflammatory response by inhibiting cardiomyocytes.Ultimately,NSLP-Val improved cardiac function and inhibited cardiac hypertrophy and fibrosis.
文摘BACKGROUND Heart failure(HF)is a global health problem characterized by impaired heart function.Cardiac remodeling and cell death contribute to the development of HF.Although treatments such as digoxin and angiotensin receptor blocker drugs have been used,their effectiveness in reducing mortality is uncertain.Researchers are exploring the use of adipose-derived mesenchymal stem cell(ADMSC)exosomes(Exos)as a potential therapy for HF.These vesicles,secreted by cells,may aid in tissue repair and regulation of inflammation and immune responses.However,further investigation is needed to understand the specific role of these vesicles in HF treatment.AIM To investigate the mechanism of extracellular vesicles produced by ADMSC s in the treatment of HF.METHODS Exogenous surface markers of ADMSCs were found,and ADMSCs were cultured.RESULTS The identification of surface markers showed that the surface markers CD44 and CD29 of adipose-derived stem cells(ADSCs)were well expressed,while the surface markers CD45 and CD34 of ADSCs were negative,so the cultured cells were considered ADSCs.Western blotting detected the Exo surface marker protein,which expressed CD63 protein but did not express calnexin protein,indicating that ADSC-derived Exos were successfully extracted.CONCLUSION The secretion of MSCs from adipose tissue can increase ATP levels,block cardiomyocyte apoptosis,and enhance the heart function of animals susceptible to HF.The inhibition of Bax,caspase-3 and p53 protein expression may be related to this process.
基金supported by the National Science Foundation of China(82130011,81770261,91749128,and 81900233)the Fundamental Research Funds for the Central Universities(2042022kf1125,China)the Outstanding Young and Middleaged Talents Training Program of Zhongnan Hospital of Wuhan University(ZNYQ2022002,China).
文摘Clinical application of doxorubicin(DOX)is heavily hindered by DOX cardiotoxicity.Several theories were postulated for DOX cardiotoxicity including DNA damage and DNA damage response(DDR),although the mechanism(s)involved remains to be elucidated.This study evaluated the potential role of TBC domain family member 15(TBC1D15)in DOX cardiotoxicity.Tamoxifen-induced cardiac-specific Tbcldi5 knockout(Tbcldi5^(CKO))or Tbcldi5 knockin(Tbcldi5^(CKI))male mice were challenged with a single dose of DOx prior to cardiac assessment 1 week or 4 weeks following DOX challenge.Adenoviruses encoding TBC1D15 or containing shRNA targeting Tbcld15 were used for Tbcld15 overexpression or knockdown in isolated primary mouse cardiomyocytes.Our results re-vealed that DOX evoked upregulation of TBC1D15 with compromised myocardial function and overt mortality,the effects of which were ameliorated and accentuated by Tbcldi5 deletion and Tbcld15 overexpression,respectively.DOX overtly evoked apoptotic cell death,the effect of which was alleviated and exacerbated by Tbcld15 knockout and overexpression,respectively.Meanwhile,DOX provoked mitochondrial membrane potential collapse,oxidative stress and DNA damage,the effects of which were mitigated and exacerbated by Tbcld15 knockdown and overexpression,respectively.Further scrutiny revealed that TBC1D15 fostered cytosolic accumulation of the cardinal DDR element DNA-dependent protein kinase catalytic subunit(DNA-PKcs).Liquid chromatography-tandem mass spectrometry and coimmunoprecipitation denoted an interaction between TBCID15 and DNA-PKcs at the segment 594-624 of TBC1D15.Moreover,overexpression of TBC1D15 mutant(A594-624,deletion of segment 594-624)failed to elicit accentuation of DOX-induced cytosolic retention of DNA-PKcs,DNA damage and cardiomyocyte apoptosis by TBC1D15 wild type.However,Tbcld15 deletion ameliorated DOXinduced cardiomyocyte contractile anomalies,apoptosis,mitochondrial anomalies,DNA damage and cytosolic DNA-PKcs accumulation,which were canceled off by DNA-PKcs inhibition or ATM activation.Taken together,our findings denoted a pivotal role for TBCID15 in DOX-induced DNA damage,mitochondrial injury,and apoptosis possibly through binding with DNA-PKcs and thus gate-keeping its cytosolic retention,a route to accentuation of cardiac contractile dysfunction in DOX-induced cardiotoxicity.
基金supported by grants from the National Natural Science Foundation of China (81872860, 81803521, 81673433)National Major Special Projects for the Creation and Manufacture of New Drugs (2019ZX09301104, China)+5 种基金Local Innovative and Research Teams Project of Guangdong Pearl River Talents Program (2017BT01Y093, China)National Engineering and Technology Research Center for New drug Druggability Evaluation(Seed Program of Guangdong Province, 2017B090903004,China)Natural Science Foundation of Guangdong Province(2019A1515010273, China)Foundation from Guangdong Traditional Medicine Bureau (20191060, China)Fundamental Research Funds for the Central Universities (19ykpy131, China)Research and Industrialization team of Taxus chinensis var.mairel (2014YT02S044, China)。
文摘As an effective anticancer drug, the clinical limitation of doxorubicin(Dox) is the time-and dose-dependent cardiotoxicity. Yes-associated protein 1(YAP1) interacts with transcription factor TEA domain 1(TEAD1) and plays an important role in cell proliferation and survival. However, the role of YAP1 in Dox-induced cardiomyopathy has not been reported. In this study, the expression of YAP1 was reduced in clinical human failing hearts with dilated cardiomyopathy and Dox-induced in vivo and in vitro cardiotoxic model. Ectopic expression of Yap1 significantly blocked Dox-induced cardiomyocytes apoptosis in TEAD1 dependent manner. Isorhapontigenin(Isor) is a new derivative of stilbene and responsible for a wide range of biological processes. Here, we found that Isor effectively relieved Doxinduced cardiomyocytes apoptosis in a dose-dependent manner in vitro. Administration with Isor(30 mg/kg/day, intraperitoneally, 3 weeks) significantly protected against Dox-induced cardiotoxicity in mice. Interestingly, Isor increased Dox-caused repression in YAP1 and the expression of its target genes in vivo and in vitro. Knockout or inhibition of Yap1 blocked the protective effects of Isor on Dox-induced cardiotoxicity. In conclusion, YAP1 may be a novel target for Dox-induced cardiotoxicity and Isor might be a new compound to fight against Dox-induced cardiotoxicity by increasing YAP1 expression.
