Kruppel样因子15对大鼠心脏压力超/卸负荷后脂质沉积及心功能的调节作用
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摘要
目的通过建立大鼠胸骨上小切口升主动脉缩窄及松解模型,探讨心脏压力超/卸负荷后心脏脂质沉积及心功能的变化。方法选用5周龄、体质量150~200 g雄性SD大鼠45只,行非人工通气下胸骨上小切口升主动脉缩窄术致大鼠心脏功能受损,造模成功后分为9组(n=5):假手术对照组(9、12、15周)分别标记为C1、C2、C3,升主动脉缩窄组(缩窄9、12、15周)分别标记为A1、A2、A3,去缩窄组(缩窄9周后去缩窄当天、3、6周组)分别标记为B1、B2、B3。通过动物超声心动图观察心脏功能,PCR和Western blot检测Kruppel样因子15(kruppel-like factor 15,KLF15),心肌脂质染色观察脂质沉积分别在A1后去缩窄,B1、B2、B3以及A1、A2、A3的变化情况。结果 1与C1、C2、C3相比,升主动脉缩窄组随着缩窄时间越长,心功能越低,A1、A2、A3与C1、C2、C3比较EF分别为:[(85.32±3.90)vs(54.32±3.88),(85.45±4.26)vs(47.91±3.64),(82.83±4.93)vs(40.08±4.45),P<0.01];2与A1相比,B2、B3心功能有所恢复,并具有统计学差异(P<0.05);EF%:(54.32±3.88)vs(63.28±5.81),(54.32±3.88)vs(63.59±6.48);3与C1、C2、C3相比,A1、B2、B3心脏KLF15 mRNA及其蛋白表达降低[mRNA:(0.66±0.09)vs(1.76±0.07),(0.52±0.07)vs(1.78±0.12),(0.51±0.04)vs(1.70±0.13),P<0.01];蛋白:[(0.99±0.05)vs(1.25±0.04),(0.96±0.05)vs(1.30±0.09),(1.01±0.08)vs(1.30±0.06),P<0.01];4与A1相比,B2、B3 KLF15 mRNA及其蛋白表达无明显变化。5与C1、C2、C3相比,A1、B2、B3心脏脂质沉积增多;与A1相比,B2、B3心脏脂质沉积有所减少,但仍明显高于对照组。结论在心脏超/卸负荷中,KLF15可能对心脏脂质沉积具有负向调节作用,参与了卸负荷后心功能的恢复过程。
Objective To determine lipid deposition and cardiac function in pressure overload /unload myocardium in rats through banding and debanding of the ascending aorta through small suprasternal incision. Methods Forty-five 5-week-old male SD rats were randomly divided into sham operation group( control group,in 9,12 and 15 weeks after small suprasternal incision respectively as C1,C2 and C3groups),ascending aortic banding groups( banding for 9,12 and 15 weeks respectively as A1,A2 and A3groups),and ascending aortic banding groups( debanding for 0,3 and 6 weeks respectively after 9 weeks' banding as B1,B2 and B3 groups),and there were 5 animals in each group. Animal ultrasonography was used to evaluate the cardiac function in each group. The expression of Kruppel-like factor 15( KLF15) at mRNA and protein levels was detected by PCR and Western blotting respectively. Lipid staining was employed to measure the lipid deposition in the myocardium during pressure overload / unload. Results Compared with the control group,the cardiac function was decreased with the increase of banding time,and the ejection fraction( EF%) was significantly lower in the A1,A2 and A3 groups than the C1,C2 and C3 groups( 54. 32 ±3. 88 vs 85. 32 ± 3. 90,47. 91 ± 3. 64 vs 85. 45 ± 4. 26,40. 08 ± 4. 45 vs 82. 83 ± 4. 93,P < 0. 01),and it was also decreased in the B1,B2 and B3 groups( 53. 78 ± 3. 58,63. 28 ± 5. 81 and 63. 59 ± 6. 48,P <0. 05). Compared to the A1 group,the EF% was significant increased in the B2 and B3( both P < 0. 05).The mRNA and protein levels of KLF15 were significant decreased in the A1,B2 and B3 groups than the C1,C2 and C3 groups( mRNA: 0. 66 ± 0. 09 vs 1. 76 ± 0. 07,0. 52 ± 0. 07 vs 1. 78 ± 0. 12,0. 51 ± 0. 04 vs1. 70 ± 0. 13; Protein: 0. 99 ± 0. 05 vs 1. 25 ± 0. 04,0. 96 ± 0. 05 vs 1. 30 ± 0. 09,1. 01 ± 0. 08 vs 1. 30 ±0. 06,all P < 0. 01). But there were no statistical differences in the levels in the A1 group with the B2 and B3 groups. Myocardial lipid deposition was higher in the A1,B2 and B3 groups than the C1,C2 and C3 groups,and its was decreased in the B2 and B3 than the A1 group,but still higher than the control group.Conclusion During cardiac pressure overload / unload,KLF15 may play negative regulative role in the lipid deposition in the heart,and is involved in cardiac function recovery after unloading.
Objective To determine lipid deposition and cardiac function in pressure overload /unload myocardium in rats through banding and debanding of the ascending aorta through small suprasternal incision. Methods Forty-five 5-week-old male SD rats were randomly divided into sham operation group( control group,in 9,12 and 15 weeks after small suprasternal incision respectively as C1,C2 and C3groups),ascending aortic banding groups( banding for 9,12 and 15 weeks respectively as A1,A2 and A3groups),and ascending aortic banding groups( debanding for 0,3 and 6 weeks respectively after 9 weeks' banding as B1,B2 and B3 groups),and there were 5 animals in each group. Animal ultrasonography was used to evaluate the cardiac function in each group. The expression of Kruppel-like factor 15( KLF15) at mRNA and protein levels was detected by PCR and Western blotting respectively. Lipid staining was employed to measure the lipid deposition in the myocardium during pressure overload / unload. Results Compared with the control group,the cardiac function was decreased with the increase of banding time,and the ejection fraction( EF%) was significantly lower in the A1,A2 and A3 groups than the C1,C2 and C3 groups( 54. 32 ±3. 88 vs 85. 32 ± 3. 90,47. 91 ± 3. 64 vs 85. 45 ± 4. 26,40. 08 ± 4. 45 vs 82. 83 ± 4. 93,P < 0. 01),and it was also decreased in the B1,B2 and B3 groups( 53. 78 ± 3. 58,63. 28 ± 5. 81 and 63. 59 ± 6. 48,P <0. 05). Compared to the A1 group,the EF% was significant increased in the B2 and B3( both P < 0. 05).The mRNA and protein levels of KLF15 were significant decreased in the A1,B2 and B3 groups than the C1,C2 and C3 groups( mRNA: 0. 66 ± 0. 09 vs 1. 76 ± 0. 07,0. 52 ± 0. 07 vs 1. 78 ± 0. 12,0. 51 ± 0. 04 vs1. 70 ± 0. 13; Protein: 0. 99 ± 0. 05 vs 1. 25 ± 0. 04,0. 96 ± 0. 05 vs 1. 30 ± 0. 09,1. 01 ± 0. 08 vs 1. 30 ±0. 06,all P < 0. 01). But there were no statistical differences in the levels in the A1 group with the B2 and B3 groups. Myocardial lipid deposition was higher in the A1,B2 and B3 groups than the C1,C2 and C3 groups,and its was decreased in the B2 and B3 than the A1 group,but still higher than the control group.Conclusion During cardiac pressure overload / unload,KLF15 may play negative regulative role in the lipid deposition in the heart,and is involved in cardiac function recovery after unloading.
引文
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[13]Gullace G,Demicheli G,Monte I,et al.Reclassification of echocardiography according to the appropriateness of use,function-and competence-based profiles and application[J].J Cardiovasc Echograph,2012,22(3):91-98.
[14]Gray S,Feinberg M W,Hull S,et al.The Kruppel-like factor KLF15 regulates the insulin-sensitive glucose transporter GLUT4[J].J Biol Chem,2002,277(37):34322-34328.
[15]Fisch S,Gray S,Heymans S,et al.Kruppel-like factor 15is a regulator of cardiomyocyte hypertrophy[J].Proc Natl Acad Sci USA,2007,104(17):7074-7079.
[16]Wang B,Haldar S M,Lu Y,et al.The Kruppel-like factor KLF15 inhibits connective tissue growth factor(CTGF)expression in cardiac fibroblasts[J].J Mol Cell Cardiol,2008,45(2):193-197.
[17]余杨,邹书帆,马杰,等.KLF15基因对心肌纤维化的抑制作用[J].第三军医大学学报,2014,36(14):1448-1453.
[18]Nagare T,Sakaue H,Matsumoto M,et al.Overexpression of KLF15 transcription factor in adipocytes of mice results in down-regulation of SCD1 protein expression in adipocytes and consequent enhancement of glucose-induced insulin secretion[J].J Biol Chem,2011,286(43):37458-37469.
[2]Xu R,Lin F,Zhang S,et al.Signal pathways involved in reverse remodeling of the hypertrophic rat heart after pressure unloading[J].Int J Cardiol,2010,143(3):414-423.
[3]Gray S,Wang B,Orihuela Y,et al.Regulation of gluconeogenesis by Kruppel-like factor 15[J].Cell Metab,2007,5(4):305-312.
[4]Haldar S M,Jeyaraj D,Anand P,et al.Kruppel-like factor15 regulates skeletal muscle lipid flux and exercise adaptation[J].Proc Natl Acad Sci U S A,2012,109(17):6739-6744.
[5]Jeyaraj D,Scheer F A,Ripperger J A,et al.Klf15 orchestrates circadian nitrogen homeostasis[J].Cell Metab,2012,15(3):311-323.
[6]Prosdocimo D A,Anand P,Liao X,et al.Kruppel-like factor 15 is a critical regulator of cardiac lipid metabolism[J].J Biol Chem,2014,289(9):5914-5924.
[7]Jung D Y,Chalasani U,Pan N,et al.KLF15 is a molecular link between endoplasmic reticulum stress and insulin resistance[J].PLo S One,2013,8(10):e77851.
[8]Haldar S M,Lu Y,Jeyaraj D,et al.Klf15 deficiency is a molecular link between heart failure and aortic aneurysm formation[J].Sci Transl Med,2010,2(26):26ra26.
[9]Razeghi P,Young M E,Alcorn J L,et al.Metabolic gene expression in fetal and failing human heart[J].Circulation,2001,104(24):2923-2931.
[10]Goldberg I J,Trent C M,Schulze P C.Lipid metabolism and toxicity in the heart[J].Cell Metab,2012,15(6):805-812.
[11]马杰,余杨,蹇召,等.胸骨上小切口大鼠主动脉瓣上缩窄模型的建立[J].中华实验外科杂志,2014,31(5):1147-1149.
[12]Nakamura A,Rokosh D G,Paccanaro M,et al.LV systolic performance improves with development of hypertrophy after transverse aortic constriction in mice[J].Am J Physiol Heart Circ Physiol,2001,281(3):H1104-H1112.
[13]Gullace G,Demicheli G,Monte I,et al.Reclassification of echocardiography according to the appropriateness of use,function-and competence-based profiles and application[J].J Cardiovasc Echograph,2012,22(3):91-98.
[14]Gray S,Feinberg M W,Hull S,et al.The Kruppel-like factor KLF15 regulates the insulin-sensitive glucose transporter GLUT4[J].J Biol Chem,2002,277(37):34322-34328.
[15]Fisch S,Gray S,Heymans S,et al.Kruppel-like factor 15is a regulator of cardiomyocyte hypertrophy[J].Proc Natl Acad Sci USA,2007,104(17):7074-7079.
[16]Wang B,Haldar S M,Lu Y,et al.The Kruppel-like factor KLF15 inhibits connective tissue growth factor(CTGF)expression in cardiac fibroblasts[J].J Mol Cell Cardiol,2008,45(2):193-197.
[17]余杨,邹书帆,马杰,等.KLF15基因对心肌纤维化的抑制作用[J].第三军医大学学报,2014,36(14):1448-1453.
[18]Nagare T,Sakaue H,Matsumoto M,et al.Overexpression of KLF15 transcription factor in adipocytes of mice results in down-regulation of SCD1 protein expression in adipocytes and consequent enhancement of glucose-induced insulin secretion[J].J Biol Chem,2011,286(43):37458-37469.