RAS阻滞剂及LCZ696对动脉粥样硬化小鼠ACE2-Ang-(1-7)-Mas轴的影响研究
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摘要
目的探讨肾素-血管紧张素系统(RAS)阻滞剂及LCZ696对动脉粥样硬化小鼠ACE2-Ang-(1-7)-Ma s轴的影响及可能的机制。方法采用6周龄健康雄性ApoE~(-/-)小鼠28只和C57BL/6小鼠7只。ApoE~(-/-)小鼠高脂喂养建模后随机分为4组,分别予以生理盐水、依那普利、缬沙坦和LCZ696灌胃处理,C57BL/6小鼠予以等量0.9%氯化钠溶液灌胃。干预6周后处死取血样及病理标本,应用全自动生化分析仪测定各组血清中LDL-C、TC水平,ELISA法测定血清CRP、IL-6、ACE2、Ang-(1-7)、AngII水平,油红O染色法测斑块脂质含量,RT-qPCR法测定主动脉ACE2 mRNA表达,免疫组化法测定主动脉斑块中ACE2和Ang-(1-7)蛋白表达。结果与C57BL/6组相比,ApoE~(-/-)对照组LDL-C、TC、CRP、IL-6水平升高(P<0.01),主动脉斑块明显增加(P<0.01)。与Ap oE~(-/-)对照组相比,依那普利组、缬沙坦组和LCZ696组血清CRP、IL-6、AngII水平显著降低(P<0.01),ACE2、Ang-(1-7)水平显著升高(P<0.01),依那普利组、缬沙坦组和LCZ696组斑块显著减少(P<0.01),依那普利组和LCZ696组的主动脉中ACE2 mRNA表达增加(P<0.01或0.05),依那普利组、缬沙坦组和LCZ696组主动脉斑块中ACE2和Ang-(1-7)蛋白表达增加(P<0.01)。与依那普利组、缬沙坦组相比,LCZ696组斑块减少更明显(P<0.05),主动脉斑块中ACE2和Ang-(1-7)蛋白表达增加更明显(P<0.01或0.05)。结论依那普利、缬沙坦和LCZ696均可以通过ACE2-Ang-(1-7)-Mas轴来抑制动脉粥样硬化的发展,LCZ696抗动脉粥样硬化作用更明显。
Objective To investigate the effect of RAS blockers and LCZ696 on ACE2-Ang-(1-7)-Mas axis in atherosclerotic mice and the possible mechanisms. Methods 28 healthy male ApoE~(-/-)mice aged 6 weeks were fed high-fat diet and randomly divided into 4 groups after animal model established and then treated with saline, Enalapril, Valsartan and LCZ696, respectively. 7C57BL/6 mice were given equal amount of saline. After six-week intervention,all mice were killed and the blood samples and pathological specimens were taken for measurements of serum LDL-C, TC by automatic biochemical analyzer, and serum levels of CRP,IL-6,ACE2, Ang-(1-7)and AngII by ELISA. The lipid content of plaque was measured by Oil Red O staining. The expression of aortic ACE2 m RNA was determined by RT-qPCR. The expression of ACE2 and Ang-(1-7) protein in aortic plaques was determined by immunohistochemistry. Results Compared with C57BL/6 group, LDL-C,TC,CRP and IL-6 as well as aortic plaque size significantly increased in ApoE~(-/-)control group(all P<0.01). Compared with ApoE~(-/-)control group, serum CRP,IL-6 and AngII decreased, serum ACE2 and Ang-(1-7)increased and the plaque decreased significantly in Enalapril, Valsartan and LCZ696 groups(all P<0.01), ACE2 m RNA expression increased in the aorta in enalapril group and LCZ696 group(P<0.01 or 0.05), and ACE2,Ang-(1-7) protein expression increased in aortic plaques in enalapril, valsartan and LCZ696 groups(P <0.01). Compared with enalapril and valsartan groups, the reduction of aortic plaque was more significant in LCZ696 group(P<0.05)and the expression of ACE2 and Ang-(1-7)protein in aortic plaque increased more(P<0.01 or 0.05). Conclusion Enalapril, valsartan and LCZ696 can all inhibit the development of atherosclerosis through ACE2-Ang-(1-7)-Mas axis, and LCZ696 has more obvious anti-atherosclerosis effect.
Objective To investigate the effect of RAS blockers and LCZ696 on ACE2-Ang-(1-7)-Mas axis in atherosclerotic mice and the possible mechanisms. Methods 28 healthy male ApoE~(-/-)mice aged 6 weeks were fed high-fat diet and randomly divided into 4 groups after animal model established and then treated with saline, Enalapril, Valsartan and LCZ696, respectively. 7C57BL/6 mice were given equal amount of saline. After six-week intervention,all mice were killed and the blood samples and pathological specimens were taken for measurements of serum LDL-C, TC by automatic biochemical analyzer, and serum levels of CRP,IL-6,ACE2, Ang-(1-7)and AngII by ELISA. The lipid content of plaque was measured by Oil Red O staining. The expression of aortic ACE2 m RNA was determined by RT-qPCR. The expression of ACE2 and Ang-(1-7) protein in aortic plaques was determined by immunohistochemistry. Results Compared with C57BL/6 group, LDL-C,TC,CRP and IL-6 as well as aortic plaque size significantly increased in ApoE~(-/-)control group(all P<0.01). Compared with ApoE~(-/-)control group, serum CRP,IL-6 and AngII decreased, serum ACE2 and Ang-(1-7)increased and the plaque decreased significantly in Enalapril, Valsartan and LCZ696 groups(all P<0.01), ACE2 m RNA expression increased in the aorta in enalapril group and LCZ696 group(P<0.01 or 0.05), and ACE2,Ang-(1-7) protein expression increased in aortic plaques in enalapril, valsartan and LCZ696 groups(P <0.01). Compared with enalapril and valsartan groups, the reduction of aortic plaque was more significant in LCZ696 group(P<0.05)and the expression of ACE2 and Ang-(1-7)protein in aortic plaque increased more(P<0.01 or 0.05). Conclusion Enalapril, valsartan and LCZ696 can all inhibit the development of atherosclerosis through ACE2-Ang-(1-7)-Mas axis, and LCZ696 has more obvious anti-atherosclerosis effect.
引文
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[11] Ras S, Sampaio WO, Alzamora AC, et al. The ACE2/Angiotensin-(1-7)/MAS Axis of the Renin-Angiotensin System∶Focus on Angiotensin-(1-7)[J]. Physiological Reviews, 2018, 98(1)∶505.DOI∶10.1152/physrev.00023.2016.
[12] Zhang YH, Hao QQ, Wang XY, et al. ACE2 activity was increased in atherosclerotic plaque by losartan:Possible relation to anti-atherosclerosis[J]. Journal of the renin-angiotensin-aldosterone system∶JRAAS, 2015, 16(2)∶292. DOI∶10.1177/1470320314542892.
[13]黄丽,应涛,何文明.不同肾素血管紧张素系统阻滞剂对ACE2-Ang-(1-7)及炎症因子的影响[J].心电与循环, 2017, 36(2)∶77-80.
[14] Mogensen UM, Kober L, Kristensen SL, et al. The effects of sacubitril/valsartan on coronary outcomes in PARADIGM-HF[J].American Heart Journal, 2017, 188∶35-41. DOI∶10.1016/j.ahj.2017.02.034.
[15] Ichiki T, Izumi R, Cataliotti A, et al. Endothelial permeability in vitro and in vivo:protective actions of ANP and omapatrilat in experimental atherosclerosis[J]. Peptides, 2013, 48(5)∶21-26.DOI∶0.1016/j.peptides.2013.07.020.
[16] Hubers SA, Brown NJ. Combined Angiotensin Receptor Antagonism and Neprilysin inhibition[J]. Circulation, 2016, 133(11)∶1115-1124. DOI∶10.1161/CIRCULATIONAHA.115.018622.
[2] Zhang F, Liu J, Li SF. Angiotensin-(1-7)∶new perspectives in atherosclerosis treatment[J].Journal of Geriatric Cardiology∶JGC,2015, 12(6)∶676-682. DOI∶10.11909/j.issn.1671-5411.2015.06.014.
[3] Solomon SD, Zile M, Pieske B, et al. The angiotensin receptor neprilysin inhibitor LCZ696 in heart failure with preserved ejection fraction:a phase 2 double-blind randomised controlled trial[J].Lancet, 2012, 380(9851)∶1387-1395. DOI∶10.1016/S0140-6736(12)61227-6.
[4] Braunwald E. The path to an Angiotensin receptor antagonistneprilysin inhibitor in the treatment of heart failure[J]. Journal of the American College of Cardiology, 2015, 65(10)∶1029. DOI∶10.1016/j.jacc.2015.01.033.
[5] Segura J, Ruilope L M. Dual-acting angiotensin receptor-neprilysin inhibition[J]. Current Hypertension Reports, 2011, 13(1)∶74.DOI∶10.1007/s11906-010-0166-7.
[6] Jin HY, Chen LJ, Zhang ZZ, et al. Deletion of angiotensin-converting enzyme 2 exacerbates renal inflammation and injury in apolipoprotein E-deficient mice through modulation of the nephrin and TNF-alpha-TNFRSF1A signaling[J]. Journal of Translational Medicine, 2015, 13(1)∶255. DOI∶10.1186/s12967-015-0616-8.
[7] Zhang F, Ren J, Chan K, et al. Angiotensin-(1-7)regulates Angiotensin II-induced VCAM-1 expression on vascular endothelial cells[J]. Biochemical&Biophysical Research Communications,2013, 430(2)∶642-646. DOI∶10.1016/j.bbrc.2012.11.098.
[8] Liang B, Wang X, Zhang N, et al. Angiotensin-(1-7)Attenuates Angiotensin II-Induced ICAM-1, VCAM-1, and MCP-1Expression via the MAS Receptor Through Suppression of P38 and NF-u03baB Pathways in HUVECs[J]. Cellular Physiology&Biochemistry, 2015, 35(6)∶2472-2482. DOI∶10.1159/000374047.
[9] Silva AR, Fraga-Silva RA, Stergiopulos N, et al. Update on the role of angiotensin in the pathophysiology of coronary atherothrombosis[J]. European Journal of Clinical Investigation, 2015, 45(3)∶274-87. DOI∶10.1111/eci.12401.
[10] Zhang YH, Zhang YH, Dong XF, et al. ACE2 and Ang-(1-7)protect endothelial cell function and prevent early atherosclerosis by inhibiting inflammatory response[J]. Inflammation Research,2015,64(3-4)∶253-260. DOI∶10.1007/s00011-015-0805-1.
[11] Ras S, Sampaio WO, Alzamora AC, et al. The ACE2/Angiotensin-(1-7)/MAS Axis of the Renin-Angiotensin System∶Focus on Angiotensin-(1-7)[J]. Physiological Reviews, 2018, 98(1)∶505.DOI∶10.1152/physrev.00023.2016.
[12] Zhang YH, Hao QQ, Wang XY, et al. ACE2 activity was increased in atherosclerotic plaque by losartan:Possible relation to anti-atherosclerosis[J]. Journal of the renin-angiotensin-aldosterone system∶JRAAS, 2015, 16(2)∶292. DOI∶10.1177/1470320314542892.
[13]黄丽,应涛,何文明.不同肾素血管紧张素系统阻滞剂对ACE2-Ang-(1-7)及炎症因子的影响[J].心电与循环, 2017, 36(2)∶77-80.
[14] Mogensen UM, Kober L, Kristensen SL, et al. The effects of sacubitril/valsartan on coronary outcomes in PARADIGM-HF[J].American Heart Journal, 2017, 188∶35-41. DOI∶10.1016/j.ahj.2017.02.034.
[15] Ichiki T, Izumi R, Cataliotti A, et al. Endothelial permeability in vitro and in vivo:protective actions of ANP and omapatrilat in experimental atherosclerosis[J]. Peptides, 2013, 48(5)∶21-26.DOI∶0.1016/j.peptides.2013.07.020.
[16] Hubers SA, Brown NJ. Combined Angiotensin Receptor Antagonism and Neprilysin inhibition[J]. Circulation, 2016, 133(11)∶1115-1124. DOI∶10.1161/CIRCULATIONAHA.115.018622.