JAK2/STAT3信号通路介导玉郎伞查尔酮后处理对大鼠离体心脏的保护作用
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摘要
目的:探讨玉郎伞查尔酮(YLSC)后处理对大鼠离体心脏缺血/再灌注损伤(I/R)的保护作用及相关信号传导通路机制。方法:将60只雄性SD大鼠随机分为正常组,模型组,YLSC组(5μmol·L~(-1)),AG490组(2μmol·L~(-1)),YLSC+AG490组(YLSC 5μmol·L~(-1)+AG490 2μmol·L~(-1)),每组12只。采用停灌30 min,再灌注120 min的方法制备大鼠离体心脏缺血/再灌注模型。再灌注前10 min给予AG490或YLSC。记录心率(HR),左心室舒张末压(LVEDP)和左室内压最大变化速率(±dp/dtmax)。收集冠脉流出液,测定冠脉流出液中乳酸脱氢酶(LDH)含量。再灌结束后处死大鼠,2,3,5-氯化三苯基四氮唑(TTC)法测定心肌梗死面积,蛋白免疫印迹法(Western blot)检测心肌组织磷酸化Janus激酶2(phosphorylated janus kinase 2,p-JAK2),磷酸化信号转导和转录激活子3(phosphorylated signal transducer and activator of transcription 3,p-STAT3),B淋巴细胞瘤-2(B-cell lymphoma-2,Bcl-2),Bcl-2相关X蛋白(Bcl-2 associated X protein,Bax)蛋白表达量。结果:与模型组比较,YLSC后处理可明显改善心功能,缩小心肌梗死面积,减少LDH含量,减少凋亡及Bax表达水平,上升p-JAK2,p-STAT3及Bcl-2表达水平(P<0.05,P<0.01)。而同时给予YLSC和AG490处理后,血流动力学改善作用减弱,心肌梗死面积,LDH及Bax表达较YLSC组回升,p-JAK2,p-STAT3,Bcl-2的表达较YLSC组降低(P<0.05,P<0.01)。结论:YLSC后处理能减轻大鼠离体心脏缺血/再灌注损伤,其机制可能与激活JAK2/STAT3通路有关。
Objective: To study the protective effect of 17-methoxyl-7-hydroxyl-benzofuran chalcone( YLSC) postconditioning on myocardial ischemia reperfusion( I/R) injury in isolated rat hearts and investigate its related signaling pathway. Method: Total 60 male SD rats were randomly divided into 5 groups: normal group,model group,YLSC group( 5 μmol·L~(-1)),AG490 group( 2 μmol·L~(-1)) and YLSC + inhibitor group( YLSC5 μmol·L~(-1)+ AG490 2 μmol·L~(-1)),n = 12 in each group. The isolated rat hearts were subjected to 30 min ischemia + 120 min reperfusion to establish the I/R model. AG490 or YLSC was given 10 min before reperfusion.The heart rate( HR),the left ventricular end diastolic pressure( LVEDP) and the maximum change rate of left ventricular pressure( ± dp/dtmax) were recorded,and the level of lactic dehydrogenase( LDH) in coronary effluent was detected. The rats were sacrificed after reperfusion, and their infarct sizes were measured by triphenyltetrazolium chloride( TTC) staining. Phosphorylated janus kinase 2( JAK2),phosphorylated signal transducer and activator of transcription 3( STAT3),B-cell lymphoma-2( Bcl-2) and Bcl-2 associated X protein( Bax) protein expression levels in myocardium were measured by Western blot. Result: As compared with the model group,YLSC postconditioning obviously ameliorated cardiac function,decreased myocardial infarct sizes and LDH level,reduced apoptotic rate and Bax expression levels,and increased p-JAK2,p-STAT3 and Bcl-2 levels( P < 0. 05,P < 0. 01). Combination of YLSC and AG490 could attenuate the hemodynamic improvement effect,increase the infarct sizes,LDH level and Bax expression level,and down-regulate p-JAK2,p-STAT3 and Bcl-2 expression levels as compared with YLSC group( P < 0. 05,P < 0. 01). Conclusion: YLSC postconditioning could effectively protect against myocardial I/R injury in isolated rat hearts,and its mechanism may be associated with activation of JAK2/STAT3 signaling pathway.
Objective: To study the protective effect of 17-methoxyl-7-hydroxyl-benzofuran chalcone( YLSC) postconditioning on myocardial ischemia reperfusion( I/R) injury in isolated rat hearts and investigate its related signaling pathway. Method: Total 60 male SD rats were randomly divided into 5 groups: normal group,model group,YLSC group( 5 μmol·L~(-1)),AG490 group( 2 μmol·L~(-1)) and YLSC + inhibitor group( YLSC5 μmol·L~(-1)+ AG490 2 μmol·L~(-1)),n = 12 in each group. The isolated rat hearts were subjected to 30 min ischemia + 120 min reperfusion to establish the I/R model. AG490 or YLSC was given 10 min before reperfusion.The heart rate( HR),the left ventricular end diastolic pressure( LVEDP) and the maximum change rate of left ventricular pressure( ± dp/dtmax) were recorded,and the level of lactic dehydrogenase( LDH) in coronary effluent was detected. The rats were sacrificed after reperfusion, and their infarct sizes were measured by triphenyltetrazolium chloride( TTC) staining. Phosphorylated janus kinase 2( JAK2),phosphorylated signal transducer and activator of transcription 3( STAT3),B-cell lymphoma-2( Bcl-2) and Bcl-2 associated X protein( Bax) protein expression levels in myocardium were measured by Western blot. Result: As compared with the model group,YLSC postconditioning obviously ameliorated cardiac function,decreased myocardial infarct sizes and LDH level,reduced apoptotic rate and Bax expression levels,and increased p-JAK2,p-STAT3 and Bcl-2 levels( P < 0. 05,P < 0. 01). Combination of YLSC and AG490 could attenuate the hemodynamic improvement effect,increase the infarct sizes,LDH level and Bax expression level,and down-regulate p-JAK2,p-STAT3 and Bcl-2 expression levels as compared with YLSC group( P < 0. 05,P < 0. 01). Conclusion: YLSC postconditioning could effectively protect against myocardial I/R injury in isolated rat hearts,and its mechanism may be associated with activation of JAK2/STAT3 signaling pathway.
引文
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[14]WANG Z,YU J,WU J,et al.Scutellarin protects cardiomyocyte ischemia-reperfusion injury by reducing apoptosis and oxidative stress[J].Life Sci,2016,157:200-207.
[15]Hattori R,Maulik N,Otani H,et al.Role of STAT3 in ischemic preconditioning[J].J Mol Cell Cardiol,2001,33(11):1929-1936.
[16]Boengler K,Hilfiker Kleiner D,Heusch G,et al.Inhibition of permeability transition pore opening by mitochondrial STAT3 and its role in myocardial ischemia/reperfusion[J].Basic Res Cardiol,2010,105(6):771-785.
[17]Szczepanek K,CHEN Q,Derecka M,et al.Mitochondrial-targeted signal transducer and activator of transcription 3(STAT3)protects against ischemiainduced changes in the electron transport chain and the generation of reactive oxygen species[J].J Biol Chem,2011,286(34):29610-29620.
[18]Bolli R,Stein A B,GUO Y R,et al.A murine model of inducible,cardiac-specific deletion of STAT3:its use to determine the role of STAT3 in the upregulation of cardioprotective proteins by ischemic preconditioning[J].J Mol Cell Cardiol,2011,50(4):589-597.
[2]Huffman L C,Koch S E,Butler K L.Coronary effluent from a preconditioned heart activates the JAK-STAT pathway and induces cardioprotection in a donor heart[J].Am J Physiol Heart Circ Physiol,2008,294(1):257-262.
[3]XIE X J,FAN D M,XI K,et al.Suppression of microRNA-135b-5p protects against myocardial ischemia/reperfusion injury by activating JAK2/STAT3signaling pathway in mice during sevoflurane anesthesia[J].Biosci Rep,2017,37(3):186-196.
[4]广西壮族自治区卫生厅.广西中药材标准[M].南宁:广西科学技术出版社,1992:31-32.
[5]简洁,张士军,邱莉,等.壮药玉郎伞查尔酮类成分的研究[J].中国医院药学杂志,2010,30(20):1734-1737.
[6]覃斐章.17-甲氧基-7-羟基-苯并呋喃查尔酮对心肌缺血的保护作用及机制研究[D].南宁:广西医科大学,2012.
[7]胡明珠,周波,盛琼,等.PI3K/Akt/Sirt1信号通路介导硫化氢后处理对大鼠缺血心肌的保护作用[J].中国药理学通报,2016,32(2):268-273.
[8]禤霏霏,黄建春,唐静芝,等.玉郎伞查尔酮调控PI3K/Akt信号通路抗心肌缺血/再灌注损伤的作用及机制研究[J].中国药理学通报,2015,31(12):1730-1734.
[9]黄建春.17-甲氧基-7-羟基-苯并呋喃查尔酮逆转压力超负荷大鼠心血管重构的作用及机制研究[D].南宁:广西医科大学,2014.
[10]Levy D E,Darnel J E.Stats:transcriptional control and biological impact[J].Nat Rev Mol Cell Biol,2002,137(3):651-662.
[11]Hilfiker K D,Hilfiker A,Fuchs M,et al.Signal transducer and activator of transcription 3 is required for myocardial capillary growth,control of interstitial matrix deposition,and heart protection from ischemic injury[J].Circ Res,2004,95(2):187-195.
[12]Kunisada K,Negoro S,Tone E,et al.Signal transducer and activator of transcription 3 in the heart transduces not only a hypertrophic signal but a protective signal against doxorubicin-induced cardiomyopathy[J].P Natl Acad Sci USA,2000,97(1):315-319.
[13]Barry S P,Townsend P A,Latchman D S,et al.Role of the JAK/STAT pathway in myocardial injury[J].Trends Mol Med,2007,13(2):82-89.
[14]WANG Z,YU J,WU J,et al.Scutellarin protects cardiomyocyte ischemia-reperfusion injury by reducing apoptosis and oxidative stress[J].Life Sci,2016,157:200-207.
[15]Hattori R,Maulik N,Otani H,et al.Role of STAT3 in ischemic preconditioning[J].J Mol Cell Cardiol,2001,33(11):1929-1936.
[16]Boengler K,Hilfiker Kleiner D,Heusch G,et al.Inhibition of permeability transition pore opening by mitochondrial STAT3 and its role in myocardial ischemia/reperfusion[J].Basic Res Cardiol,2010,105(6):771-785.
[17]Szczepanek K,CHEN Q,Derecka M,et al.Mitochondrial-targeted signal transducer and activator of transcription 3(STAT3)protects against ischemiainduced changes in the electron transport chain and the generation of reactive oxygen species[J].J Biol Chem,2011,286(34):29610-29620.
[18]Bolli R,Stein A B,GUO Y R,et al.A murine model of inducible,cardiac-specific deletion of STAT3:its use to determine the role of STAT3 in the upregulation of cardioprotective proteins by ischemic preconditioning[J].J Mol Cell Cardiol,2011,50(4):589-597.