丹参酮ⅡA对急性心肌梗死大鼠心脏功能和心肌线粒体自噬的影响
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摘要
目的:探究丹参酮ⅡA(TSⅡA)对急性心肌梗死(AMI)大鼠心脏功能的作用及机制。方法:将60只大鼠随机分为对照(Ctrl)组、心肌梗死(MI)组、TSⅡA(20)组、TSⅡA(50)组和TSⅡA(100)组。除对照组外,其余组采用冠状动脉结扎术建立急性心肌梗死模型,TSⅡA(20)组、TSⅡA(50)组和TSⅡA(100 mg)组腹腔注射20 mg/kg、50 mg/kg和100 mg/kg的TSⅡA,其余两组给予生理盐水。21 d后检测大鼠平均动脉压(MAP)、心率(HR)和左心室收缩压(LVSP),并分离心肌,计算心肌梗死面积,HE染色检测心肌病理损伤,Tunel染色检测细胞凋亡,试剂盒检测血清超氧化物歧化酶(SOD)、丙二醛(MDA)和谷胱甘肽(GSH)浓度,Western blot检测肌红蛋白(Mb)、肌酸激酶同工酶(CK-MB)、肌酐蛋白Ⅰ(c TnⅠ)及Beclin 1、P62和LC3的表达。结果:与对照组比较,心肌梗死组大鼠MAP、HR和LVSP均明显降低,心肌组织Mb、CK-MB和c TnⅠ表达明显增多;与心肌梗死组比较,TSⅡA(20、50、100)组大鼠MAP、HR和LVSP均明显升高,Mb、CK-MB和c TnⅠ蛋白表达明显被抑制。同时,心肌梗死组大鼠心肌损伤与对照组比较明显加重,细胞凋亡明显增多,心肌梗死面积均明显增大; TSⅡA(20、50、100)组大鼠心肌损伤情况与心肌梗死组比较明显减轻,凋亡细胞明显减少,心肌梗死面积明显减小。此外,心肌梗死能显著降低大鼠血清SOD和GSH浓度,升高MDA浓度,还能显著升高LC3Ⅱ/LC3Ⅰ比值和Beclin 1的表达水平,抑制P62表达; TSⅡA(20、50、100)能明显诱导SOD和GSH分泌,降低血清MDA浓度,还能显著下调LC3Ⅱ/LC3Ⅰ比值和Beclin 1的表达水平,促进P62表达。结论:丹参酮ⅡA(TSⅡA)可能通过抑制心肌线粒体自噬促进心肌梗死大鼠心肌功能的恢复。
Objective: To investigate the effects and mechanisms of tanshinone ⅡA( TSⅡA) on cardiac function of myocardial infraction rats. Methods: 60 rats were divided into control,myocardial infraction( MI),TSⅡA( 20 mg),TSⅡA( 50 mg) and TSⅡA( 100 mg) groups. The coronary artery of rats was ligated for establishing the MI model except control group. The rats in TS Ⅱ A( 20 mg),TS Ⅱ A( 50 mg) and TS Ⅱ A( 100 mg) groups were injected intraperitoneally with TS Ⅱ A( 20 mg/kg,50 mg/kg and100 mg/kg) respectively,and others were treated with saline. Three weeks after rats treating with TSⅡA,the mean arterial pressure( MAP),heat rate( HR) and left ventricular systolic pressure( LVSP) were measured. The myocardial tissues were removed and calculated myocardial infraction area of rats,the pathological injury was determined by HE staining. Tunel assay was performed for apoptosis,the concentrations of superoxide dismutase( SOD),malondialdehyde( MDA) and glutathione( GSH) were measured by kits. The expressions of myoglobin( Mb),creatine kinase MB( CK-MB),cardiac troponin Ⅰ( c TnⅠ),Beclin 1,P62 and LC3 were determined by Western blot. Results: Compared with control group,the MAP,HR,LVSP and the expression levels of Mb,CK-MB and c TnⅠwere up-regulated; compared with MI group,the MAP,HR,LVSP and the expression levels of Mb,CK-MB and c TnⅠwere down-regulated in TSⅡA( 20,50,100) group markedly. Meanwhile,the pathological injury was aggravated in MI group compared with Ctrl group,the cell apoptosis was induced and myocardial infraction area of rats in MI group were increased significantly. The injury of myocardial tissue was alleviated in TSⅡA( 20,50,100) group compared with myocardial infraction group,cell apoptosis was inhibited and infraction size of rats in TSⅡA( 20,50,100) group were decreased. In addition,MI inhibited secretions of SOD and GSH,but induced secretion of MDA,and up-regulated the ratio of LC3Ⅱ/LC3Ⅰand protein levels of Beclin 1,inhibited the expression of P62. TSⅡA( 20,50,100) induced the sections of SOD,GSH and expression of P62 in MI rats,but decreased the concentration of MDA,the ratio of LC3Ⅱ/LC3Ⅰand protein level of Belcin 1. Conclusion: Tanshinone ⅡA( TSⅡA) may promote the recovery of cardiac function of myocardial infraction rats by inhibiting mitophagy.
Objective: To investigate the effects and mechanisms of tanshinone ⅡA( TSⅡA) on cardiac function of myocardial infraction rats. Methods: 60 rats were divided into control,myocardial infraction( MI),TSⅡA( 20 mg),TSⅡA( 50 mg) and TSⅡA( 100 mg) groups. The coronary artery of rats was ligated for establishing the MI model except control group. The rats in TS Ⅱ A( 20 mg),TS Ⅱ A( 50 mg) and TS Ⅱ A( 100 mg) groups were injected intraperitoneally with TS Ⅱ A( 20 mg/kg,50 mg/kg and100 mg/kg) respectively,and others were treated with saline. Three weeks after rats treating with TSⅡA,the mean arterial pressure( MAP),heat rate( HR) and left ventricular systolic pressure( LVSP) were measured. The myocardial tissues were removed and calculated myocardial infraction area of rats,the pathological injury was determined by HE staining. Tunel assay was performed for apoptosis,the concentrations of superoxide dismutase( SOD),malondialdehyde( MDA) and glutathione( GSH) were measured by kits. The expressions of myoglobin( Mb),creatine kinase MB( CK-MB),cardiac troponin Ⅰ( c TnⅠ),Beclin 1,P62 and LC3 were determined by Western blot. Results: Compared with control group,the MAP,HR,LVSP and the expression levels of Mb,CK-MB and c TnⅠwere up-regulated; compared with MI group,the MAP,HR,LVSP and the expression levels of Mb,CK-MB and c TnⅠwere down-regulated in TSⅡA( 20,50,100) group markedly. Meanwhile,the pathological injury was aggravated in MI group compared with Ctrl group,the cell apoptosis was induced and myocardial infraction area of rats in MI group were increased significantly. The injury of myocardial tissue was alleviated in TSⅡA( 20,50,100) group compared with myocardial infraction group,cell apoptosis was inhibited and infraction size of rats in TSⅡA( 20,50,100) group were decreased. In addition,MI inhibited secretions of SOD and GSH,but induced secretion of MDA,and up-regulated the ratio of LC3Ⅱ/LC3Ⅰand protein levels of Beclin 1,inhibited the expression of P62. TSⅡA( 20,50,100) induced the sections of SOD,GSH and expression of P62 in MI rats,but decreased the concentration of MDA,the ratio of LC3Ⅱ/LC3Ⅰand protein level of Belcin 1. Conclusion: Tanshinone ⅡA( TSⅡA) may promote the recovery of cardiac function of myocardial infraction rats by inhibiting mitophagy.
引文
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[14]Chen Z,Xu H.Anti-inflammatory and immunomodulatory mechanism of tanshinoneⅡA for atherosclerosis[J].Evid Based Complement Alternat Med,2014,2014:267976.
[15]Ren ZH,Tong YH,Xu W,et al.Tanshinone II A attenuates inflammatory responses of rats with myocardial infarction by reducing MCP-1 expression[J].Phytomedicine,2010,17(3-4):212-218.
[16]Tan S,Zhou S,Luo Y.Baicalein pretreatment confers cardioprotection against acute myocardial infarction by activating the endothelial nitric oxide synthase signaling pathway and inhibiting oxidative stress[J].Mol Med Rep,2014,9(6):2429-2434.
[17]Pipaliya H,Vaghasiya J.Cardio protective effect of vitamin A against isoproterenol-induced myocardial infarction[J].J Nutr Sci Vitaminol,2012,58(6):402-407.
[18]Wang N,Chang Y,Chen L,et al.TanshinoneⅡA protects against chronic intermittent hypoxia-induced myocardial injury via activating the endothelin 1 pathway[J].Biomed Pharmacother,2017,95:1013-1020.
[19]Feng Y,Zhao J,Hou H,et al.WDR26 promotes mitophagy of cardiomyocytes induced by hypoxia through Parkin translocation[J].Acta Biochim Biophys Sin,2016,48(12):1075-1084.
[20]Lin WJ,Kuang HY.Oxidative stress induces autophagy in response,to multiple noxious stimuli in retinal ganglion cells[J].Autophagy,2014,10(10):1692-1701.
[21]Dianat M,Sadeghi N,Badavi M,et al.Protective effects of co-administration of gallic acid and cyclosporine on rat myocardial morphology against ischemia/reperfusion[J].Jundishapur J Nat Pharm Prod,2014,9(4):e17186.
[22]Jia Z,Lin L,Huang S,et al.Inhibition of autophagy by berberine enhances the survival of H9C2 myocytes following hypoxia[J].Mol Med Rep,2017,16(2):1677-1684.
[23]Zhu Y,Tang Q,Wang G,et al.TanshinoneⅡA protects hippocampal neuronal cells from reactive oxygen species through changes in autophagy and activation of phosphatidylinositol 3-kinase,protein kinas b,and mechanistic target of rapamycin pathways[J].Curr Neurovasc Res,2017,14(2):132-140.
[24]Pan CC,Kumar S,Shah N,et al.Endoglin regulation of Smad2function mediates Beclin1 expression and endothelial autophagy[J].J Biol Chem,2015,290(24):14884-14892.
[25]Jiang P,Mizushima N.LC3-and p62-based biochemical methods for the analysis of autophagy progression in mammalian cells[J].Methods,2015,75:13-18.
[2]Seropian IM,Toldo S,Tassell BWV,et al.Anti-inflammatory strategies for ventricular remodeling following ST-segment elevation acute myocardial infarction[J].J Am Coll Cardiol,2014,63(16):1593-1603.
[3]Zhao BY,Liao ZF,Chen S,et al.Intramyocardial transplantation of cardiac telocytes decreases myocardial infarction and improves post-infarcted cardiac function in rats[J].J Cell Mol Med,2014,18(5):780-789.
[4]Kubli DA,Zhang X,Lee Y,et al.Parkin protein deficiency exacerbates cardiac injury and reduces survival following myocardial infarction[J].J Biol Chem,2013,288(2):915-926.
[5]Queliconi BB,Kowaltowski AJ,Gottlieb RA.Bicarbonate increases ischemia-reperfusion damage by inhibiting mitophagy[J].PLo SOne,2016,11(12):e0167678.
[6]班努·库肯,钟小兰,严金龙.大鼠心肌梗死后心脏线粒体及自噬小体的变化与Parkin蛋白活性的相关性[J].临床心血管病杂志,2016,6:598-602.Bannu·Kuken,Zhong XL,Yan JL.Activity of Parkin protein in the changes of heart mitochondria and autophagosome after myocardial infraction in rats[J].J Clin Cardiol,2016,6:598-602.
[7]Sui X,Gao C.Huperzine A ameliorates damage induced by acute myocardial infarction in rats through antioxidant,anti-apoptotic and anti-inflammatory mechanisms[J].Int J Mol Med,2014,33(1):227-233.
[8]Chen P,Pang S,Yang N,et al.Beneficial effects of schisandrin Bon the cardiac function in mice model of myocardial infarction[J].PLo S One,2013,8(8):e79418.
[9]Yuan X,Jing S,Wu L,et al.Pharmacological postconditioning with tanshinoneⅡA attenuates myocardial ischemia-reperfusion injury in rats by activating the phosphatidylinositol 3-kinase pathway[J].Exp Ther Med,2014,8(3):973-977.
[10]Queliconi BB,Wojtovich AP,Nadtochiy SM,et al.Redox regulation of the mitochondrial K(ATP)channel in cardioprotection[J].Biochim Biophys Acta,2011,1813(7):1309-1315.
[11]Hu H,Zhai C,Qian G,et al.Protective effects of tanshinoneⅡAon myocardial ischemia reperfusion injury by reducing oxidative stress,HMGB1 expression,and inflammatory reaction[J].Pharm Biol,2015,53(12):1752-1758.
[12]夏金华,夏建川,谢丽燕,等.丹参酮ⅡA对树突状细胞表型的影响及对功能的调控[J].中国免疫学杂志,2017,33(3):374-377.Xia JH,Xia JC,Xie LY,et al.A novel role for tanshinoneⅡA in modulation of dendritic cells maturation and function[J].Chin JImmunol,2017,33(3):374-377.
[13]Hu Q,Wei B,Wei L,et al.Sodium tanshinoneⅡA sulfonate ameliorates ischemia-induced myocardial inflammation and lipid accumulation in Beagle dogs through NLRP3 inflammasome[J].Int J Cardiol,2015,196:183-192.
[14]Chen Z,Xu H.Anti-inflammatory and immunomodulatory mechanism of tanshinoneⅡA for atherosclerosis[J].Evid Based Complement Alternat Med,2014,2014:267976.
[15]Ren ZH,Tong YH,Xu W,et al.Tanshinone II A attenuates inflammatory responses of rats with myocardial infarction by reducing MCP-1 expression[J].Phytomedicine,2010,17(3-4):212-218.
[16]Tan S,Zhou S,Luo Y.Baicalein pretreatment confers cardioprotection against acute myocardial infarction by activating the endothelial nitric oxide synthase signaling pathway and inhibiting oxidative stress[J].Mol Med Rep,2014,9(6):2429-2434.
[17]Pipaliya H,Vaghasiya J.Cardio protective effect of vitamin A against isoproterenol-induced myocardial infarction[J].J Nutr Sci Vitaminol,2012,58(6):402-407.
[18]Wang N,Chang Y,Chen L,et al.TanshinoneⅡA protects against chronic intermittent hypoxia-induced myocardial injury via activating the endothelin 1 pathway[J].Biomed Pharmacother,2017,95:1013-1020.
[19]Feng Y,Zhao J,Hou H,et al.WDR26 promotes mitophagy of cardiomyocytes induced by hypoxia through Parkin translocation[J].Acta Biochim Biophys Sin,2016,48(12):1075-1084.
[20]Lin WJ,Kuang HY.Oxidative stress induces autophagy in response,to multiple noxious stimuli in retinal ganglion cells[J].Autophagy,2014,10(10):1692-1701.
[21]Dianat M,Sadeghi N,Badavi M,et al.Protective effects of co-administration of gallic acid and cyclosporine on rat myocardial morphology against ischemia/reperfusion[J].Jundishapur J Nat Pharm Prod,2014,9(4):e17186.
[22]Jia Z,Lin L,Huang S,et al.Inhibition of autophagy by berberine enhances the survival of H9C2 myocytes following hypoxia[J].Mol Med Rep,2017,16(2):1677-1684.
[23]Zhu Y,Tang Q,Wang G,et al.TanshinoneⅡA protects hippocampal neuronal cells from reactive oxygen species through changes in autophagy and activation of phosphatidylinositol 3-kinase,protein kinas b,and mechanistic target of rapamycin pathways[J].Curr Neurovasc Res,2017,14(2):132-140.
[24]Pan CC,Kumar S,Shah N,et al.Endoglin regulation of Smad2function mediates Beclin1 expression and endothelial autophagy[J].J Biol Chem,2015,290(24):14884-14892.
[25]Jiang P,Mizushima N.LC3-and p62-based biochemical methods for the analysis of autophagy progression in mammalian cells[J].Methods,2015,75:13-18.