Aim： To observe effects of angiotensin （Ang） II receptor antagonist （ATI） irbesartan and angiotensin-converting enzyme （ACE） inhibitor perindopril on rat myocardium calcineurin expression and sarcoplasmic retic...Aim： To observe effects of angiotensin （Ang） II receptor antagonist （ATI） irbesartan and angiotensin-converting enzyme （ACE） inhibitor perindopril on rat myocardium calcineurin expression and sarcoplasmic reticulum Ca^2＋-ATPase activity in the model of pressure-overload cardiac hypertrophy. Methods： Forty male adult Sprague Dawley rats were divided into 5 groups One group was treated by sham operation; four groups were myocardium hypertrophy cases caused by banding aortic above renal artery. Drugs were given one week after operation. Group 1： sham group, rats （n=8） were gavaged with normal saline 2 ml/（kg·d） （ig）; Group 2： control group, rats （n=8） were treated with normal saline 2 ml/（kg·d） （ig）; Group 3： rats （n=8） were given perindopril 2 mg/（kg·d） （ig）; Group 4： rats （n=8） were treated with irbesartan 20 mg/（kg·d） （ig）; Group 5： rats （n=8） were given irbesartan 20 mg/（kg·d） plus perindopril 2 mg/（kg·d） （ig）. Morphometric determination, calcineurin expression and sarcoplasmic reticulum Ca^2＋-ATPase activity were done at the end of 6 week of drug intervention. Expression of calcineurin in myocardium was detected by immunohistochemistry. Results： Left ventricular mass index （LVMI）, transverse diameter of myocardial cell （TDM）, calcineurin activity were remarkably decreased after drug intervention and this decrease was most remarkable in the combination drug therapy group. Sarcoplasmic reticulum Ca^2＋-ATPase activity was increased after drug intervention, especially in the combined drug therapy group. Calcineurin expression in myocardium was remarkably decreased after drug intervention. LVMI was positively correlated with TDM and calcineurin, negatively correlated with sarcoplasmic reticulum Ca^2＋-ATPase. Conclusion： These data suggest that irbesartan and perindopril inhibit cardiac hypertrophy through the increased activity of sarcoplasmic reticulum Ca^2＋-ATPase and decreased expression of calcineurin. Their combination had better effects on regressing of ventricular hypertrophy.展开更多
Objective To detect changes in the calcium pump protein or the calcium release channel of thesarcoplasmic reticulum during chronic doxorubicin treatment. Methods The rats were treated with intravenousdoxorubicin(lmg/k...Objective To detect changes in the calcium pump protein or the calcium release channel of thesarcoplasmic reticulum during chronic doxorubicin treatment. Methods The rats were treated with intravenousdoxorubicin(lmg/kg) twice weekly for 12 to 18 times. Controls received intravenous normal saline. The seventy ofcardiomyopathy was scored by light and electron microscopic study to investigate left ventricular papillary muscleand the calcium handling of the myocardial sarcoplasmic reticulum (SR) was determined using the isotope45 Ca2+loading. Results The ability of SR Ca2+ uptake was decreased in doxorubicin- treated rats compared withcontrol rats and the magnitude of the decrease in SR Ca2+ uptake was correlated with the seventy of thecardiomyopathy graded by pathology score. The percentage of the SR calcium release decreased 14.3% ± 4.2% in theinitial 10s and decreased 17.1 %± 4.5% (P<0.05) at 2min in the severe groups as compared with control (P<0.01)and the amount of SR calcium release seemed correlate with the seventy of the cardiomyopathy graded.Conclusion The altered function of SR calcium uptake and release could lead to the abnormalities of contractionand relaxation observed in the doxorubicin cardiomyopathy.展开更多
Objective To study the effect of phospholamban antisense RNA （asPLB） on sarcoplasmic reticulum Ca2＋-ATPase activity and cardiac function in rats with diabetes mellitus （DM） mediated by recombinant adeno-associate...Objective To study the effect of phospholamban antisense RNA （asPLB） on sarcoplasmic reticulum Ca2＋-ATPase activity and cardiac function in rats with diabetes mellitus （DM） mediated by recombinant adeno-associated virus （rAAV） vector. Methods Six weeks after the induction of DM by streptozotocin injected intraperitoneally, the rats were divided into three groups, namely： DM-rAAV-asPLB group, DM-saline group and DM group （control group）. The rats in the DM-rAAV-asPLB group were intramyocardially injected with rAAV-asPLB, the rats in the DM-saline group were injected with saline, and those in the control group did not receive any treatment. Six weeks after gene transfer, the expressions of PLB protein and PLB phosphorylation were detected by Western-blot, while the activity of sarcoplasmic reticulum （SR） Ca2＋-ATPase and left ventricular function were measured. Results The PLB protein expression level was significantly higher whereas the PLB phosphorylation, SR Ca2＋-ATPase activity and left ventricular function were significantly lower in the DM-saline group than in the control group. No significant difference was found in PLB protein expression level, PLB phosphorylation or SR Ca2＋-ATPase activity between the DM-rAAV-asPLB group and the control group. The left ventricular function in the DM-rAAV-asPLB group was poorer than in the control group and was better than in the DM-saline group. Conclusion rAAV-asPLB can down-regulate PLB protein expression and up-regulate phosphorylation and SR Ca2＋-ATPase activity, thus contributing to the improvement of in vivo ventricutar function. PLB left展开更多
目的探讨去甲基化酶脂肪质量和肥胖相关基因(fat mass and obesity associated gene,FTO)在糖尿病冠脉平滑肌收缩功能异常中的影响。方法Cre-loxP重组技术制备平滑肌特异性FTO敲除小鼠(FTO^(SMKO))。分为4组:对照组(WT)、糖尿病组(DM)、...目的探讨去甲基化酶脂肪质量和肥胖相关基因(fat mass and obesity associated gene,FTO)在糖尿病冠脉平滑肌收缩功能异常中的影响。方法Cre-loxP重组技术制备平滑肌特异性FTO敲除小鼠(FTO^(SMKO))。分为4组:对照组(WT)、糖尿病组(DM)、FTO敲除组(FTO^(SMKO))和FTOSMKO糖尿病组(FTO^(SMKO)-DM),每组各15只。糖尿病小鼠由腹腔注射链脲佐菌素(STZ)制备;其余小鼠注射等量的柠檬酸-柠檬酸钠缓冲液。通过小血管环张力测定技术,观察5-HT对4组小鼠冠脉平滑肌收缩反应的影响;采用Western blot与Dot blot技术检测小鼠血管组织FTO蛋白及n6-甲基腺嘌呤(m6A)甲基化修饰水平的变化。结果与WT组相比,DM组血糖明显升高(P<0.01),体质量明显下降(P<0.05);DM组小鼠主动脉FTO蛋白水平升高( P <0.01),m6A甲基化修饰水平降低( P <0.01)。DM组5-HT诱导收缩反应与WT组相比明显下降( P <0.01),而FTOSMKO-DM组收缩反应比DM组明显增加( P <0.01);FTO^(SMKO) -DM组非L型钙通道介导的血管平滑肌收缩反应增强,其中,1,4,5-三磷酸肌醇受体(IP3R)和咖啡因激活兰尼碱受体(RyR)介导的肌浆网钙释放诱导收缩反应均明显增加( P <0.05)。 结论 特异性敲除平滑肌 FTO 可改善糖尿病小鼠冠脉对血管收缩剂5-HT的反应性,可能与FTO介导5-HT受体信号通路异常有关。展开更多
Theaflavins from black tea effectively improve insulin secretion in obesity and diabetes,but the molecular mechanisms are unclear.Here,the palmitic acid(PA)-induced pancreaticβ-TC-6 cells and high fat-/high glucose-i...Theaflavins from black tea effectively improve insulin secretion in obesity and diabetes,but the molecular mechanisms are unclear.Here,the palmitic acid(PA)-induced pancreaticβ-TC-6 cells and high fat-/high glucose-induced zebrafish were used.The effects of theaflavin-3,3'-digallate(TF3)on glucolipotoxicityinduced insulin secretion dysfunction,ferroptosis and endoplasmic reticulum(ER)stress were investigated by a variety of molecular biological approaches,inductively coupled plasma-mass spectrometry(ICP-MS),transmission electron microscopy(TEM)and widely targeted metabolomics analysis.TF3 was found to potently inhibit glucolipotoxicity-induced insulin secretion dysfunction and ferroptosis inβ-TC-6 cells and zebrafish,with increasing glutathione peroxidase 4(GPX4)expression,suppressing lipid peroxidation and iron accumulation and protecting mitochondria.Additionally,TF3 attenuated ER stress by regulating 3 unfolded protein response(UPR)pathways inβ-TC-6 cells,and significantly modulated linoleic acid metabolism and L-kynurenine signalling in zebrafish.The expression of sarcoplasmic/endoplasmic reticulum calcium ATPase 2(SERCA2)was obviously enhanced by TF3.Thapsigargin,a SERCA2 inhibitor,remarkably reversed the effects of TF3 on insulin production,ferroptosis,ER stress and the kynurenine signalling.Together,this work revealed the critical role of SERCA2 in ferroptosis regulation,and demonstrated TF3 targeted SERCA2 to inhibit ER stress and ferropto sis,thereby protectingβ-cell secretory function from glucolipotoxicity.展开更多
BACKGROUND Myocardial infarction(MI)is a significant global cause of chronic heart failure.In post-ischemic cardiac hypertrophy,multiple molecular targets and signals within the cardiac tissue are evident.Mesenchymal ...BACKGROUND Myocardial infarction(MI)is a significant global cause of chronic heart failure.In post-ischemic cardiac hypertrophy,multiple molecular targets and signals within the cardiac tissue are evident.Mesenchymal stem cell-derived exosomes(MSC-EXO)and exercise(EXE)showed promise in enhancing post-ischemic cardiac repair.AIM To investigate how the exosomes released by stem cells and/or EXE can promote cardiac repair and improve isoproterenol(ISO)-induced post-ischemic hypertrophy.METHODS The enrolled animals were divided into 8 control rats and 32 experimental rats.Induction of MI was performed using ISO.Then,the experimental rats were divided into 4 groups:Rats subjected to 4 weeks of swimming EXE,rats treated with exosomes,and the combined treatment.Additionally,functional and interactional exploration of targeted proteins was conducted using Gene Ontology,Kyoto Encyclopedia of Genes and Genomes analysis,and STRING database,along with histological examination.RESULTS Both MSC-EXO or EXE significantly improved ISO induced elevation of cardiac enzymes,oxidative stress,and inflammatory markers,as well as the degenerative changes of the cardiac muscles,fibrosis,and apoptosis.Meanwhile,the combined treatment of EXE and MSC-EXO resulted in a significant improvement in cardiac function and structure as compared to all groups that synchronized with dual inhibition of extracellular signal-regulated kinase and protein kinase B/mammalian target of rapamycin(P<0.01)signaling and modulation of matrix metalloproteinase 9 and sarcoplasmic endoplasmic reticulum calcium ATPase type 2a,with significant improved angiogenesis.CONCLUSION Functional and structural cardiac improvements are accompanied by reduced inflammation,oxidative stress,and apoptosis.Both MSC-EXO and EXE exert cardio-protection by upregulating sarcoplasmic endoplasmic reticulum calcium ATPase,the critical pump for normal calcium handling.展开更多
目的:研究扩心方对阿霉素诱导扩张型心肌病模型大鼠心功能、心肌损伤的保护作用,探讨扩心方作用机制。方法:50只Wistar大鼠腹腔注射阿霉素6周建立DCM模型,开始造模4周后随机分为模型组,扩心方高、中、低剂量组,卡托普利组(每组各10只),...目的:研究扩心方对阿霉素诱导扩张型心肌病模型大鼠心功能、心肌损伤的保护作用,探讨扩心方作用机制。方法:50只Wistar大鼠腹腔注射阿霉素6周建立DCM模型,开始造模4周后随机分为模型组,扩心方高、中、低剂量组,卡托普利组(每组各10只),连续灌胃给药四周,另设10只正常对照。正常组和模型组以生理盐水灌胃。治疗结束后,对各组大鼠进行心脏超声检查,病理形态学检测,以及用反转录-聚合酶链反应和蛋白质免疫印迹法分别检测心肌组织中SERCA2α、PLB的m RNA和蛋白含量。结果:与正常对照组相比,模型组心功能减弱,出现心肌病理损害,SERCA2α与PLB表达异常;与模型组相比,扩心方组可以改善DCM大鼠的心功能、心肌病理学形态,同时,提高SERCA2αm RNA和蛋白表达、抑制PLB m RNA和蛋白表达。结论:扩心方能提高DCM大鼠的左心室射血分数和缩短率,减轻心肌损伤,改善心功能,同时调控DCM大鼠SERCA2α、PLB的表达。提示扩心方改善DCM大鼠心功能的作用机制,可能与纠正心肌肌浆网钙调控相关蛋白-SERCA2α、PLB的异常表达相关。展开更多
文摘Aim： To observe effects of angiotensin （Ang） II receptor antagonist （ATI） irbesartan and angiotensin-converting enzyme （ACE） inhibitor perindopril on rat myocardium calcineurin expression and sarcoplasmic reticulum Ca^2＋-ATPase activity in the model of pressure-overload cardiac hypertrophy. Methods： Forty male adult Sprague Dawley rats were divided into 5 groups One group was treated by sham operation; four groups were myocardium hypertrophy cases caused by banding aortic above renal artery. Drugs were given one week after operation. Group 1： sham group, rats （n=8） were gavaged with normal saline 2 ml/（kg·d） （ig）; Group 2： control group, rats （n=8） were treated with normal saline 2 ml/（kg·d） （ig）; Group 3： rats （n=8） were given perindopril 2 mg/（kg·d） （ig）; Group 4： rats （n=8） were treated with irbesartan 20 mg/（kg·d） （ig）; Group 5： rats （n=8） were given irbesartan 20 mg/（kg·d） plus perindopril 2 mg/（kg·d） （ig）. Morphometric determination, calcineurin expression and sarcoplasmic reticulum Ca^2＋-ATPase activity were done at the end of 6 week of drug intervention. Expression of calcineurin in myocardium was detected by immunohistochemistry. Results： Left ventricular mass index （LVMI）, transverse diameter of myocardial cell （TDM）, calcineurin activity were remarkably decreased after drug intervention and this decrease was most remarkable in the combination drug therapy group. Sarcoplasmic reticulum Ca^2＋-ATPase activity was increased after drug intervention, especially in the combined drug therapy group. Calcineurin expression in myocardium was remarkably decreased after drug intervention. LVMI was positively correlated with TDM and calcineurin, negatively correlated with sarcoplasmic reticulum Ca^2＋-ATPase. Conclusion： These data suggest that irbesartan and perindopril inhibit cardiac hypertrophy through the increased activity of sarcoplasmic reticulum Ca^2＋-ATPase and decreased expression of calcineurin. Their combination had better effects on regressing of ventricular hypertrophy.
文摘Objective To detect changes in the calcium pump protein or the calcium release channel of thesarcoplasmic reticulum during chronic doxorubicin treatment. Methods The rats were treated with intravenousdoxorubicin(lmg/kg) twice weekly for 12 to 18 times. Controls received intravenous normal saline. The seventy ofcardiomyopathy was scored by light and electron microscopic study to investigate left ventricular papillary muscleand the calcium handling of the myocardial sarcoplasmic reticulum (SR) was determined using the isotope45 Ca2+loading. Results The ability of SR Ca2+ uptake was decreased in doxorubicin- treated rats compared withcontrol rats and the magnitude of the decrease in SR Ca2+ uptake was correlated with the seventy of thecardiomyopathy graded by pathology score. The percentage of the SR calcium release decreased 14.3% ± 4.2% in theinitial 10s and decreased 17.1 %± 4.5% (P<0.05) at 2min in the severe groups as compared with control (P<0.01)and the amount of SR calcium release seemed correlate with the seventy of the cardiomyopathy graded.Conclusion The altered function of SR calcium uptake and release could lead to the abnormalities of contractionand relaxation observed in the doxorubicin cardiomyopathy.
基金supported by Scientific Research Foundation of College of Medicine,Zhejiang UniversityFoundation of Zhejiang Science and Technology Bureau(No.2007B071)+1 种基金Foundation of Zhejiang Educational Committee(No.Y200908338)Foundation of Zhejiang Traditional Chinese Medicine Committee(No.2011B503103)
文摘Objective To study the effect of phospholamban antisense RNA （asPLB） on sarcoplasmic reticulum Ca2＋-ATPase activity and cardiac function in rats with diabetes mellitus （DM） mediated by recombinant adeno-associated virus （rAAV） vector. Methods Six weeks after the induction of DM by streptozotocin injected intraperitoneally, the rats were divided into three groups, namely： DM-rAAV-asPLB group, DM-saline group and DM group （control group）. The rats in the DM-rAAV-asPLB group were intramyocardially injected with rAAV-asPLB, the rats in the DM-saline group were injected with saline, and those in the control group did not receive any treatment. Six weeks after gene transfer, the expressions of PLB protein and PLB phosphorylation were detected by Western-blot, while the activity of sarcoplasmic reticulum （SR） Ca2＋-ATPase and left ventricular function were measured. Results The PLB protein expression level was significantly higher whereas the PLB phosphorylation, SR Ca2＋-ATPase activity and left ventricular function were significantly lower in the DM-saline group than in the control group. No significant difference was found in PLB protein expression level, PLB phosphorylation or SR Ca2＋-ATPase activity between the DM-rAAV-asPLB group and the control group. The left ventricular function in the DM-rAAV-asPLB group was poorer than in the control group and was better than in the DM-saline group. Conclusion rAAV-asPLB can down-regulate PLB protein expression and up-regulate phosphorylation and SR Ca2＋-ATPase activity, thus contributing to the improvement of in vivo ventricutar function. PLB left
文摘目的探讨去甲基化酶脂肪质量和肥胖相关基因(fat mass and obesity associated gene,FTO)在糖尿病冠脉平滑肌收缩功能异常中的影响。方法Cre-loxP重组技术制备平滑肌特异性FTO敲除小鼠(FTO^(SMKO))。分为4组:对照组(WT)、糖尿病组(DM)、FTO敲除组(FTO^(SMKO))和FTOSMKO糖尿病组(FTO^(SMKO)-DM),每组各15只。糖尿病小鼠由腹腔注射链脲佐菌素(STZ)制备;其余小鼠注射等量的柠檬酸-柠檬酸钠缓冲液。通过小血管环张力测定技术,观察5-HT对4组小鼠冠脉平滑肌收缩反应的影响;采用Western blot与Dot blot技术检测小鼠血管组织FTO蛋白及n6-甲基腺嘌呤(m6A)甲基化修饰水平的变化。结果与WT组相比,DM组血糖明显升高(P<0.01),体质量明显下降(P<0.05);DM组小鼠主动脉FTO蛋白水平升高( P <0.01),m6A甲基化修饰水平降低( P <0.01)。DM组5-HT诱导收缩反应与WT组相比明显下降( P <0.01),而FTOSMKO-DM组收缩反应比DM组明显增加( P <0.01);FTO^(SMKO) -DM组非L型钙通道介导的血管平滑肌收缩反应增强,其中,1,4,5-三磷酸肌醇受体(IP3R)和咖啡因激活兰尼碱受体(RyR)介导的肌浆网钙释放诱导收缩反应均明显增加( P <0.05)。 结论 特异性敲除平滑肌 FTO 可改善糖尿病小鼠冠脉对血管收缩剂5-HT的反应性,可能与FTO介导5-HT受体信号通路异常有关。
基金supported by National Natural Science Foundation of China(32272303)Natural Science Foundation of Zhejiang Province,China(LY21C200010)。
文摘Theaflavins from black tea effectively improve insulin secretion in obesity and diabetes,but the molecular mechanisms are unclear.Here,the palmitic acid(PA)-induced pancreaticβ-TC-6 cells and high fat-/high glucose-induced zebrafish were used.The effects of theaflavin-3,3'-digallate(TF3)on glucolipotoxicityinduced insulin secretion dysfunction,ferroptosis and endoplasmic reticulum(ER)stress were investigated by a variety of molecular biological approaches,inductively coupled plasma-mass spectrometry(ICP-MS),transmission electron microscopy(TEM)and widely targeted metabolomics analysis.TF3 was found to potently inhibit glucolipotoxicity-induced insulin secretion dysfunction and ferroptosis inβ-TC-6 cells and zebrafish,with increasing glutathione peroxidase 4(GPX4)expression,suppressing lipid peroxidation and iron accumulation and protecting mitochondria.Additionally,TF3 attenuated ER stress by regulating 3 unfolded protein response(UPR)pathways inβ-TC-6 cells,and significantly modulated linoleic acid metabolism and L-kynurenine signalling in zebrafish.The expression of sarcoplasmic/endoplasmic reticulum calcium ATPase 2(SERCA2)was obviously enhanced by TF3.Thapsigargin,a SERCA2 inhibitor,remarkably reversed the effects of TF3 on insulin production,ferroptosis,ER stress and the kynurenine signalling.Together,this work revealed the critical role of SERCA2 in ferroptosis regulation,and demonstrated TF3 targeted SERCA2 to inhibit ER stress and ferropto sis,thereby protectingβ-cell secretory function from glucolipotoxicity.
文摘BACKGROUND Myocardial infarction(MI)is a significant global cause of chronic heart failure.In post-ischemic cardiac hypertrophy,multiple molecular targets and signals within the cardiac tissue are evident.Mesenchymal stem cell-derived exosomes(MSC-EXO)and exercise(EXE)showed promise in enhancing post-ischemic cardiac repair.AIM To investigate how the exosomes released by stem cells and/or EXE can promote cardiac repair and improve isoproterenol(ISO)-induced post-ischemic hypertrophy.METHODS The enrolled animals were divided into 8 control rats and 32 experimental rats.Induction of MI was performed using ISO.Then,the experimental rats were divided into 4 groups:Rats subjected to 4 weeks of swimming EXE,rats treated with exosomes,and the combined treatment.Additionally,functional and interactional exploration of targeted proteins was conducted using Gene Ontology,Kyoto Encyclopedia of Genes and Genomes analysis,and STRING database,along with histological examination.RESULTS Both MSC-EXO or EXE significantly improved ISO induced elevation of cardiac enzymes,oxidative stress,and inflammatory markers,as well as the degenerative changes of the cardiac muscles,fibrosis,and apoptosis.Meanwhile,the combined treatment of EXE and MSC-EXO resulted in a significant improvement in cardiac function and structure as compared to all groups that synchronized with dual inhibition of extracellular signal-regulated kinase and protein kinase B/mammalian target of rapamycin(P<0.01)signaling and modulation of matrix metalloproteinase 9 and sarcoplasmic endoplasmic reticulum calcium ATPase type 2a,with significant improved angiogenesis.CONCLUSION Functional and structural cardiac improvements are accompanied by reduced inflammation,oxidative stress,and apoptosis.Both MSC-EXO and EXE exert cardio-protection by upregulating sarcoplasmic endoplasmic reticulum calcium ATPase,the critical pump for normal calcium handling.
文摘目的:研究扩心方对阿霉素诱导扩张型心肌病模型大鼠心功能、心肌损伤的保护作用,探讨扩心方作用机制。方法:50只Wistar大鼠腹腔注射阿霉素6周建立DCM模型,开始造模4周后随机分为模型组,扩心方高、中、低剂量组,卡托普利组(每组各10只),连续灌胃给药四周,另设10只正常对照。正常组和模型组以生理盐水灌胃。治疗结束后,对各组大鼠进行心脏超声检查,病理形态学检测,以及用反转录-聚合酶链反应和蛋白质免疫印迹法分别检测心肌组织中SERCA2α、PLB的m RNA和蛋白含量。结果:与正常对照组相比,模型组心功能减弱,出现心肌病理损害,SERCA2α与PLB表达异常;与模型组相比,扩心方组可以改善DCM大鼠的心功能、心肌病理学形态,同时,提高SERCA2αm RNA和蛋白表达、抑制PLB m RNA和蛋白表达。结论:扩心方能提高DCM大鼠的左心室射血分数和缩短率,减轻心肌损伤,改善心功能,同时调控DCM大鼠SERCA2α、PLB的表达。提示扩心方改善DCM大鼠心功能的作用机制,可能与纠正心肌肌浆网钙调控相关蛋白-SERCA2α、PLB的异常表达相关。
