CaMKⅡ和PLB在房颤心房重构中的作用及意义
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摘要
目的分析风湿性心脏病(风心病)患者右心房组织受磷蛋白(phospholamban,PLB)及其调节蛋白钙调蛋白(CaMKⅡ)的mRNA水平、蛋白及磷酸化水平,探讨CaMKⅡ和PLB在心房纤颤(房颤)与心房重构中的作用机制。方法收集本院2017年7-10月30例风心病患者右心房组织标本[男性12例,女性18例,年龄24～68(50. 8±11. 4)岁],其中窦性心律组11例,房颤心律组19例。RTPCR检测两组CaMKⅡ和PLB的mRNA,Western blot检测两组CaMKⅡp286、PLBp17和PLB的表达情况。结果与窦性心律组比较,房颤心律组CaMKⅡ(0. 006±0. 003)和PLB(1. 704±1. 008)的基因表达量均显著增加(P <0. 05),CaMKⅡp286(0. 812±0. 227)和PLBp17(0. 649±0. 354)蛋白水平显著增加(P <0. 05),而PLB蛋白水平则显著降低(0. 886±0. 364,P <0. 05)。结论心房组织内CaMKⅡ和PLB的mRNA水平以及CaMKⅡp286和PLBp17的水平与房颤呈正相关,从而PLB单体水平降低,调节PLB来调控心房肌细胞钙离子循环有望成为房颤心房重构治疗的靶点。
Objective To investigate the mRNA and protein and phosphorylation levels of phospholamban( PLB) and its regulatory protein Ca2 +/calmodulin dependent protein kinase Ⅱ( CaMKⅡ) in right atrial tissue of patients with rheumatic heart disease( RHD),and explore their roles and significance in atrial fibrillation and atrial remodeling. Methods Right atrial tissue samples from 30 patients with rheumatic heart disease [12 males and 18 females,aged 24 ～ 68( 50. 8 ± 11. 4) years] were collected from our institute during July and October 2017. The patients were assigned into the sinus rhythm group( n = 11) and the atrial fibrillation rhythm group( n = 19). The mRNA levels of Ca MKⅡ and PLB were examined by RT-PCR,and protein levels of Ca MKⅡp286,PLBp17 and PLB were measured by Western blotting. Results Compared with the sinus rhythm group,the mRNA levels of CaMK Ⅱ( 0. 006 ± 0. 003) and PLB( 1. 704 ± 1. 008) were significantly higher in the atrial fibrillation rhythm group( P < 0. 05). While,similar trends were seen in the protein levels of CaMKⅡp286( 0. 812 ± 0. 227) and PLBp17( 0. 649 ± 0. 354)( P < 0. 05),but the expression level of PLB was opposite( 0. 886 ± 0. 364,P < 0. 05). Conclusion The mRNA levels of CaMKⅡ and PLB and the protein levels of CaMKⅡp286 and PLBp17 in atrial tissues are positively correlated with severity of atrial fibrillation. Therefore,decreasing PLB monomer and thus regulating PLB to control the cycle of calcium ions in atrial myocytes might be expected to a target for atrial remodeling therapy in atrial fibrillation.
Objective To investigate the mRNA and protein and phosphorylation levels of phospholamban( PLB) and its regulatory protein Ca2 +/calmodulin dependent protein kinase Ⅱ( CaMKⅡ) in right atrial tissue of patients with rheumatic heart disease( RHD),and explore their roles and significance in atrial fibrillation and atrial remodeling. Methods Right atrial tissue samples from 30 patients with rheumatic heart disease [12 males and 18 females,aged 24 ～ 68( 50. 8 ± 11. 4) years] were collected from our institute during July and October 2017. The patients were assigned into the sinus rhythm group( n = 11) and the atrial fibrillation rhythm group( n = 19). The mRNA levels of Ca MKⅡ and PLB were examined by RT-PCR,and protein levels of Ca MKⅡp286,PLBp17 and PLB were measured by Western blotting. Results Compared with the sinus rhythm group,the mRNA levels of CaMK Ⅱ( 0. 006 ± 0. 003) and PLB( 1. 704 ± 1. 008) were significantly higher in the atrial fibrillation rhythm group( P < 0. 05). While,similar trends were seen in the protein levels of CaMKⅡp286( 0. 812 ± 0. 227) and PLBp17( 0. 649 ± 0. 354)( P < 0. 05),but the expression level of PLB was opposite( 0. 886 ± 0. 364,P < 0. 05). Conclusion The mRNA levels of CaMKⅡ and PLB and the protein levels of CaMKⅡp286 and PLBp17 in atrial tissues are positively correlated with severity of atrial fibrillation. Therefore,decreasing PLB monomer and thus regulating PLB to control the cycle of calcium ions in atrial myocytes might be expected to a target for atrial remodeling therapy in atrial fibrillation.
引文
[1] DOBREV D, CARLSSON LNATTEL S. Novel molecular targets for atrial fibrillation therapy[J]. Nat Rev Drug Discov,2012,11(4):275-291. DOI:10. 1038/nrd3682.
[2] KOURLIOUROS A,SAVELIEVA I,KIOTSEKOGLOU A,et al.Current concepts in the pathogenesis of atrial fibrillation[J].Am Heart J,2009,157(2):243-252. DOI:10. 1016/j. ahj.2008. 10. 009.
[3] LAW M L,PRINS K W,OLANDER M E,et al. Exacerbation of dystrophic cardiomyopathy by phospholamban deficiencymediated chronically increased cardiac Ca2+cycling in vivo[J]. Am J Physiol Heart Circ Physiol,2018,315(6):H1544-H1552. DOI:10. 1152/ajpheart. 00341. 2018.
[4]李勇,马瑞彦,肖颖彬,等.风湿性二尖瓣病变并发心房颤动患者483例临床分析[J].第三军医大学学报,2011,33(10):1048-1051.LI Y,MA R Y,XIAO Y B,et al. Clinical analysis of 483cases of rheumatic mitral valve disease with atrial fibrillation[J]. J Third Mil Med Univ,2011,33(10):1048-1051.
[5] JANUARY C T,WANN L S,ALPERT J S,et al. 2014AHA/ACC/HRS guideline for the management of patients with atrial fibrillation:a report of the American College of Cardiology/American Heart Association Task Force on practice guidelines and the Heart Rhythm Society[J]. Circulation,2014,130(23):e199-267. DOI:10. 1161/cir.0000000000000041.
[6] AHMAD F,SHEN W,VANDEPUT F,et al. Regulation of sarcoplasmic reticulum Ca2+ATPase 2(SERCA2)activity by phosphodiesterase 3A(PDE3A)in human myocardium:phosphorylation-dependent interaction of PDE3A1 with SERCA2[J]. J Biol Chem,2015,290(11):6763-6776.DOI:10. 1074/jbc. M115. 638585.
[7] MESUBI O O,ANDERSON M E. Atrial remodelling in atrial fibrillation:Ca MKⅡas a nodal proarrhythmic signal[J].Cardiovasc Res,2016,109(4):542-557. DOI:10. 1093/cvr/cvw002.
[8] DONG X,THOMAS D D. Time-resolved FRET reveals the structural mechanism of SERCA-PLB regulation[J]. Biochem Biophys Res Commun,2014,449(2):196-201. DOI:10.1016/j. bbrc. 2014. 04. 166.
[9] GUSTAVSSON M,VERARDI R,MULLEN D G,et al.Allosteric regulation of SERCA by phosphorylation-mediated conformational shift of phospholamban[J]. Proc Natl Acad Sci U S A,2013,110(43):17338-17343. DOI:10. 1073/pnas. 1303006110.
[10] FERNANDEZ-DE GORTARI E,ESPINOZA-FONSECA L M.Structural basis for relief of phospholamban-mediated inhibition of the sarcoplasmic reticulum Ca2+-ATPase at saturating Ca2+conditions[J]. J Biol Chem,2018,293(32):12405-12414. DOI:10. 1074/jbc. RA118. 003752.
[11] ONAL B,UNUDURTHI S D,HUND T J. Modeling Ca MKⅡin cardiac physiology:from molecule to tissue[J]. Front Pharmacol,2014,5:9. DOI:10. 3389/fphar. 2014. 00009.
[12] HEIJMAN J, VOIGT N, NATTEL S, et al. Calcium handling and atrial fibrillation[J]. Wien Med Wochenschr,2012,162(13/14):287-291. DOI:10. 1007/s10354-012-0109-9.
[13] GRAMLEY F,LORENZEN J,JEDAMZIK B,et al. Atrial fibrillation is associated with cardiac hypoxia[J]. Cardiovasc Pathol,2010,19(2):102-111. DOI:10. 1016/j. carpath. 2008. 11. 001.
[14] JALIFE J. Mechanisms of persistent atrial fibrillation[J].Curr Opin Cardiol,2014,29(1):20-27. DOI:10. 1097/HCO. 0000000000000027.
[15] CHEN M,XU D,WU A Z,et al. Phospholamban regulates nuclear Ca2+stores and inositol 1,4,5-trisphosphate mediated nuclear Ca2+cycling in cardiomyocytes[J]. J Mol Cell Cardiol,2018,123:185-197. DOI:10. 1016/j. yjmcc.2018. 09. 008.
[16] BOCCHI L,SAVI M,NAPONELLI V,et al. Long-term oral administration of theaphenon-E improves cardiomyocyte mechanics and calcium dynamics by affecting phospholamban phosphorylation and ATP production[J]. Cell Physiol Biochem,2018,47(3):1230-1243. DOI:10. 1159/000490219.
[17] OLIVEIRA L G,MORENO L G,MELO D S,et al. Caryocar brasiliense oil improves cardiac function by increasing Serca2a/PLB ratio despite no significant changes in cardiovascular risk factors in rats[J]. Lipids Health Dis,2017,16(1):37. DOI:10. 1186/s12944-017-0422-9.
[2] KOURLIOUROS A,SAVELIEVA I,KIOTSEKOGLOU A,et al.Current concepts in the pathogenesis of atrial fibrillation[J].Am Heart J,2009,157(2):243-252. DOI:10. 1016/j. ahj.2008. 10. 009.
[3] LAW M L,PRINS K W,OLANDER M E,et al. Exacerbation of dystrophic cardiomyopathy by phospholamban deficiencymediated chronically increased cardiac Ca2+cycling in vivo[J]. Am J Physiol Heart Circ Physiol,2018,315(6):H1544-H1552. DOI:10. 1152/ajpheart. 00341. 2018.
[4]李勇,马瑞彦,肖颖彬,等.风湿性二尖瓣病变并发心房颤动患者483例临床分析[J].第三军医大学学报,2011,33(10):1048-1051.LI Y,MA R Y,XIAO Y B,et al. Clinical analysis of 483cases of rheumatic mitral valve disease with atrial fibrillation[J]. J Third Mil Med Univ,2011,33(10):1048-1051.
[5] JANUARY C T,WANN L S,ALPERT J S,et al. 2014AHA/ACC/HRS guideline for the management of patients with atrial fibrillation:a report of the American College of Cardiology/American Heart Association Task Force on practice guidelines and the Heart Rhythm Society[J]. Circulation,2014,130(23):e199-267. DOI:10. 1161/cir.0000000000000041.
[6] AHMAD F,SHEN W,VANDEPUT F,et al. Regulation of sarcoplasmic reticulum Ca2+ATPase 2(SERCA2)activity by phosphodiesterase 3A(PDE3A)in human myocardium:phosphorylation-dependent interaction of PDE3A1 with SERCA2[J]. J Biol Chem,2015,290(11):6763-6776.DOI:10. 1074/jbc. M115. 638585.
[7] MESUBI O O,ANDERSON M E. Atrial remodelling in atrial fibrillation:Ca MKⅡas a nodal proarrhythmic signal[J].Cardiovasc Res,2016,109(4):542-557. DOI:10. 1093/cvr/cvw002.
[8] DONG X,THOMAS D D. Time-resolved FRET reveals the structural mechanism of SERCA-PLB regulation[J]. Biochem Biophys Res Commun,2014,449(2):196-201. DOI:10.1016/j. bbrc. 2014. 04. 166.
[9] GUSTAVSSON M,VERARDI R,MULLEN D G,et al.Allosteric regulation of SERCA by phosphorylation-mediated conformational shift of phospholamban[J]. Proc Natl Acad Sci U S A,2013,110(43):17338-17343. DOI:10. 1073/pnas. 1303006110.
[10] FERNANDEZ-DE GORTARI E,ESPINOZA-FONSECA L M.Structural basis for relief of phospholamban-mediated inhibition of the sarcoplasmic reticulum Ca2+-ATPase at saturating Ca2+conditions[J]. J Biol Chem,2018,293(32):12405-12414. DOI:10. 1074/jbc. RA118. 003752.
[11] ONAL B,UNUDURTHI S D,HUND T J. Modeling Ca MKⅡin cardiac physiology:from molecule to tissue[J]. Front Pharmacol,2014,5:9. DOI:10. 3389/fphar. 2014. 00009.
[12] HEIJMAN J, VOIGT N, NATTEL S, et al. Calcium handling and atrial fibrillation[J]. Wien Med Wochenschr,2012,162(13/14):287-291. DOI:10. 1007/s10354-012-0109-9.
[13] GRAMLEY F,LORENZEN J,JEDAMZIK B,et al. Atrial fibrillation is associated with cardiac hypoxia[J]. Cardiovasc Pathol,2010,19(2):102-111. DOI:10. 1016/j. carpath. 2008. 11. 001.
[14] JALIFE J. Mechanisms of persistent atrial fibrillation[J].Curr Opin Cardiol,2014,29(1):20-27. DOI:10. 1097/HCO. 0000000000000027.
[15] CHEN M,XU D,WU A Z,et al. Phospholamban regulates nuclear Ca2+stores and inositol 1,4,5-trisphosphate mediated nuclear Ca2+cycling in cardiomyocytes[J]. J Mol Cell Cardiol,2018,123:185-197. DOI:10. 1016/j. yjmcc.2018. 09. 008.
[16] BOCCHI L,SAVI M,NAPONELLI V,et al. Long-term oral administration of theaphenon-E improves cardiomyocyte mechanics and calcium dynamics by affecting phospholamban phosphorylation and ATP production[J]. Cell Physiol Biochem,2018,47(3):1230-1243. DOI:10. 1159/000490219.
[17] OLIVEIRA L G,MORENO L G,MELO D S,et al. Caryocar brasiliense oil improves cardiac function by increasing Serca2a/PLB ratio despite no significant changes in cardiovascular risk factors in rats[J]. Lipids Health Dis,2017,16(1):37. DOI:10. 1186/s12944-017-0422-9.
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