苦参碱对阿霉素所致H9c2心肌细胞损伤的保护作用及与线粒体Na~+-K~+-ATP酶、Ca~(2+)-ATP酶关系的研究
|
摘要
目的:研究苦参碱对阿霉素所致H9c2心肌细胞损伤的保护作用及其机制。方法:将H9c2心肌细胞培养,取2～3代细胞进行试验,共分为4组。对照组:接受生理盐水作为干预因素;阿霉素组:接受0.5 mg·L~(-1)阿霉素作为损伤模型;苦参碱+阿霉素组:接受0.5 mg·L~(-1)阿霉素和不同浓度苦参碱(50,150,200 mg·L~(-1))作为干预因素;苦参碱组:接受不同浓度苦参碱(50,150,200 mg·L~(-1))作为干预因素。以上各组相应干预24 h后,应用流式细胞仪检测H9c2心肌细胞凋亡水平,应用分光光度法检测线粒体Na~+-K~+-ATP酶、Ca~(2+)-ATP酶活性,利用JC-1法检测线粒体膜电位。结果:与阿霉素组相比,苦参碱+阿霉素组H9c2心肌细胞凋亡显著减少、线粒体Na~+-K~+-ATP酶、Ca~(2+)-ATP酶活性显著升高(P<0.05)、线粒体膜电位显著改善。结论:苦参碱对阿霉素所致H9c2心肌细胞有保护作用,减轻心肌细胞凋亡,改善线粒体膜电位、提高线粒体Na~+-K~+-ATP酶、Ca~(2+)-ATP酶活性。
OBJECTIVE To study the protective effect and mechanism of matrine on H9 c2 cardiomyocytes injury induced by adriamycin.METHODS H9 c2 cardiomyocytes were cultured and 2-3 passages of cells were used for testing.They were divided into 4 groups.Control group: received normal saline as intervention factor;adriamycin group:received 0.5 mg·L~(-1) adriamycin as injury model;matrine+adriamycin group:received 0.5 mg·L~(-1) adriamycin and different concentrations of matrine(50,150,200 mg·L~(-1)) as an intervention factor; matrine group: received different concentrations of matrine(50,150,200 mg·L~(-1)) as an intervention factor.After 24 hours of intervention,the apoptosis of H9 c2 cardiomyocytes was detected by flow cytometry.The mitochondrial Na~+-K~+-ATPase and Ca~(2+)-ATPase activities were detected by spectrophotometry.The mitochondrial membrane potential was detected by JC-1 method. RESULTS Compared with the adriamycin group,the apoptosis of H9 c2 cardiomyocytes in the matrine+adriamycin group was significantly decreased,and the mitochondrial Na~+-K~+-ATPase and Ca~(2+)-ATPase activities were significantly increased(P<0.05).Mitochondrial membrane potential is significantly improved.CONCLUSION Matrine has protective effect on H9 c2 cardiomyocytes induced by adriamycin,reduces cardiomyocyte apoptosis, improves mitochondrial membrane potential,and increases mitochondrial Na~+-K~+-ATPase and Ca~(2+)-ATPase activities.
OBJECTIVE To study the protective effect and mechanism of matrine on H9 c2 cardiomyocytes injury induced by adriamycin.METHODS H9 c2 cardiomyocytes were cultured and 2-3 passages of cells were used for testing.They were divided into 4 groups.Control group: received normal saline as intervention factor;adriamycin group:received 0.5 mg·L~(-1) adriamycin as injury model;matrine+adriamycin group:received 0.5 mg·L~(-1) adriamycin and different concentrations of matrine(50,150,200 mg·L~(-1)) as an intervention factor; matrine group: received different concentrations of matrine(50,150,200 mg·L~(-1)) as an intervention factor.After 24 hours of intervention,the apoptosis of H9 c2 cardiomyocytes was detected by flow cytometry.The mitochondrial Na~+-K~+-ATPase and Ca~(2+)-ATPase activities were detected by spectrophotometry.The mitochondrial membrane potential was detected by JC-1 method. RESULTS Compared with the adriamycin group,the apoptosis of H9 c2 cardiomyocytes in the matrine+adriamycin group was significantly decreased,and the mitochondrial Na~+-K~+-ATPase and Ca~(2+)-ATPase activities were significantly increased(P<0.05).Mitochondrial membrane potential is significantly improved.CONCLUSION Matrine has protective effect on H9 c2 cardiomyocytes induced by adriamycin,reduces cardiomyocyte apoptosis, improves mitochondrial membrane potential,and increases mitochondrial Na~+-K~+-ATPase and Ca~(2+)-ATPase activities.
引文
[1] Pecoraro M,Sorrentino R,Franceschelli S,et al.Doxorubicin-mediated cardiotoxicity:Role of mitochondrial connexin 43[J].Cardiovasc Toxicol,2015,15(4):366-376.
[2] DongJ,Shao W,Yan P,et al.Curcumolide,a unique sesquiterpenoid with anti-inflammatory properties from Curcuma wenyujin[J].Bioorg Med Chem Lett,2015,25(2):198-202.
[3] Zhi X,Chen X,Su JC.Research progress in the pharmacological action of matrine[J].Chengdu Univ Tradit Chin Med (成都中医药大学学报),2017,40(1):123-127.
[4] OctaviaY,Tocchetti CG,Gabrielson KL,et al.Doxorubicin-induced cardiomyopathy:From molecular mechanisms to therapeutic strategies[J].J Mol Cell Cardiol,2012,52:1213-1225.
[5] Carvalho C,Santos RX,Cardoso S,et al.Doxorubicin:The good,the bad and the ugly effect[J].Curr Med Chem,2009,16:3267-3285.
[6] ParkerMA,King V,Howard KP.Nuclear magnetic resonance study of doxorubicin binding to cardiolipin containing magnetically oriented phospholipid bilayers[J].Biochim Biophys Acta,2001,1514:206-216.
[7] MartinSS,Senior AE.Membrane adenosine triphosphatase activities in rat pancreas[J].Biochim Biophys Acta,1980,602:401-418.
[8] Chu EF,Lu JK,Wang YQ,et al.Protective effect and mechanism of matrine on myocardial injury induced by adriamycin in rats[J].Drugs & Clinic (现代药物与临床),2016,31(5),572-576.
[9] Xu M,Yang L,Hong LZ,Zhao XY,et al.Direct protection of neurons and astrocytes by matrine via inhibition of the NF-κB signaling pathway contributes to neuroprotection against focal cerebral ischemia[J].Brain Res,2012,1454:48-64.
[10] ZhangJ,Li Y,Chen X,et al.Autophagy is involved in anticancer effects of matrine on SGC-7901 human gastric cancer cells[J].Oncol Rep,2011,26:115-124.
[11] ZhangY,Cui L,Guan GC,et al.Matrine suppresses cardiac fibrosis by inhibiting the TGF-β/Smad pathway in experimental diabetic cardiomyopathy[J].Molecular Med Reports,2018,17:1775-1781.
[12] ZhouYH,Shan HL,Qiao GF et al.Inotropic effects and mechanisms of matrine,a main alkaloid from Sophora flavescens Ait[J].Biol Pharm Bull,2008,31:2057-2062.
[13] Ling JY,Zhang GY,CuiZJ,et al.Supercritical fluid extraction of quinolizidine alkaloids from Sophora flavescens Ait.and purification by high-speed counter-current chromatography[J].J Chromatogr A,2007,23:123-127.
[14] Lu JK,Chu EF,Tony H,et al.Curcumin downregulates phosphate carrier and protects against doxorubicin induced cardiomyocyte apoptosis[J].Bio Med Res Int Vo,2016,2016:1980763.
[2] DongJ,Shao W,Yan P,et al.Curcumolide,a unique sesquiterpenoid with anti-inflammatory properties from Curcuma wenyujin[J].Bioorg Med Chem Lett,2015,25(2):198-202.
[3] Zhi X,Chen X,Su JC.Research progress in the pharmacological action of matrine[J].Chengdu Univ Tradit Chin Med (成都中医药大学学报),2017,40(1):123-127.
[4] OctaviaY,Tocchetti CG,Gabrielson KL,et al.Doxorubicin-induced cardiomyopathy:From molecular mechanisms to therapeutic strategies[J].J Mol Cell Cardiol,2012,52:1213-1225.
[5] Carvalho C,Santos RX,Cardoso S,et al.Doxorubicin:The good,the bad and the ugly effect[J].Curr Med Chem,2009,16:3267-3285.
[6] ParkerMA,King V,Howard KP.Nuclear magnetic resonance study of doxorubicin binding to cardiolipin containing magnetically oriented phospholipid bilayers[J].Biochim Biophys Acta,2001,1514:206-216.
[7] MartinSS,Senior AE.Membrane adenosine triphosphatase activities in rat pancreas[J].Biochim Biophys Acta,1980,602:401-418.
[8] Chu EF,Lu JK,Wang YQ,et al.Protective effect and mechanism of matrine on myocardial injury induced by adriamycin in rats[J].Drugs & Clinic (现代药物与临床),2016,31(5),572-576.
[9] Xu M,Yang L,Hong LZ,Zhao XY,et al.Direct protection of neurons and astrocytes by matrine via inhibition of the NF-κB signaling pathway contributes to neuroprotection against focal cerebral ischemia[J].Brain Res,2012,1454:48-64.
[10] ZhangJ,Li Y,Chen X,et al.Autophagy is involved in anticancer effects of matrine on SGC-7901 human gastric cancer cells[J].Oncol Rep,2011,26:115-124.
[11] ZhangY,Cui L,Guan GC,et al.Matrine suppresses cardiac fibrosis by inhibiting the TGF-β/Smad pathway in experimental diabetic cardiomyopathy[J].Molecular Med Reports,2018,17:1775-1781.
[12] ZhouYH,Shan HL,Qiao GF et al.Inotropic effects and mechanisms of matrine,a main alkaloid from Sophora flavescens Ait[J].Biol Pharm Bull,2008,31:2057-2062.
[13] Ling JY,Zhang GY,CuiZJ,et al.Supercritical fluid extraction of quinolizidine alkaloids from Sophora flavescens Ait.and purification by high-speed counter-current chromatography[J].J Chromatogr A,2007,23:123-127.
[14] Lu JK,Chu EF,Tony H,et al.Curcumin downregulates phosphate carrier and protects against doxorubicin induced cardiomyocyte apoptosis[J].Bio Med Res Int Vo,2016,2016:1980763.