心肌自噬在异丙肾上腺素诱导心肌肥厚中的作用
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摘要
目的探讨心肌自噬在异丙肾上腺素(ISO)诱导大鼠心肌肥厚中的作用。方法将29只大鼠随机分为3组:皮下联合腹腔注射生理盐水(Con)组9只、腹腔注射生理盐水联合皮下注射异丙肾上腺(ISO)组10只、腹腔注射雷帕霉素(RAP)联合皮下注射ISO(ISO+RAP)组10只。连续干预14 d后,将Con组和ISO组存活大鼠(Con,n=9;ISO,n=8)行心脏超声检查左室壁厚度及左室射血分数(LVEF),再将各组大鼠腹腔注射过量的10%水合氯醛处死,摘除心脏,计算心脏体质量指数(HWI)。心肌组织病理切片行HE染色和Masson染色,Western blot检测p62表达和LC3Ⅱ/Ⅰ比值,透射电镜观察心肌组织自噬泡数量。结果与Con组比较,ISO组左室壁厚度和LVEF明显增加(P<0.01)。与Con组比较,ISO组和ISO+RAP组的HWI增大(P<0.01),且ISO+RAP组的HWI小于ISO组(P<0.01)。与Con组比较,ISO组心肌病理切片显示心肌组织病理性肥厚改变明显;而与ISO组比较,ISO+RAP组心肌组织病理性肥厚明显改善。与Con组比较,ISO组心肌组织LC3Ⅱ/Ⅰ比值下调,而p62表达上调,自噬泡数量明显减少(P<0.05);与ISO组比较,ISO+RAP组心肌组织LC3Ⅱ/Ⅰ比值上调,而p62表达下调,自噬泡数量明显增多(P<0.01)。结论上调心肌自噬活性可以逆转ISO诱导的病理性心肌肥厚。
Objective To explore the effects of myocardial autophagy on isoproterenol(ISO)-induced cardiac pathological hypertrophy in rats.MethodsTwenty-nine SD rats were randomly divided into three groups: subcutaneous injection of normal saline combined with intraperitoneal injection of normal saline(Con,n=9) group,subcutaneous injection of normal saline combined with intraperitoneal injection of isoproterenol(ISO,n=10) group and subcutaneous injection of rapamycin(RAP) combined with intraperitoneal injection of ISO(ISO+RAP,n=10) group. After 14-days of continuous treatment,cardiac echocardiogram for left ventricular wall thicknesses and ejection fraction(LVEF) were carried out in Con group(n=9) and ISO group(n=8). Rats were executed by intraperitoneal injection of overdoes 10% chloral hydrate. Heart to body index(HWI) was calculated by heart weight/body weight. Cardiac pathologic alternation was observed by histological sections stained with hematoxylin and eosin(HE) and Masson. The expression of p62 and the ratio of LC3Ⅱ/Ⅰ were measured by Western blot assay. The number of autophagic vacuoles was detected by transmission electron microscopy.ResultsCompared with Con group,the left ventricular wall thickness and LVEF were significantly increased in ISO group(P<0.01). Compared with Con group,the HWI was increased in ISO group and ISO+RAP group(P<0.01). But the HWI was decreased in ISO+RAP group comparison with that of ISO group(P<0.01). Compared with Con group,the alternation of cardiac pathological hypertrophy was marked in ISO group. Moreover,compared with ISO group,cardiac pathological hypertrophy was improved in ISO+RAP group. Compared with Con group,the ratio of LC3Ⅱ/Ⅰwas down-regulated,the protein expression of p62 was up-regulated and the number of autophagic vesicles was significantly reduced in ISO group(P<0.01). However,compared with ISO group,the ratio of LC3Ⅱ/Ⅰwas up-regulated,the protein expression of p62 was down-regulated and the number of autophagic vesicles was significantly increased in ISO + RAP group(P<0.01).ConclusionThe up-regulation of myocardial autophagy can reverse pathological hypertrophy induced by ISO.
Objective To explore the effects of myocardial autophagy on isoproterenol(ISO)-induced cardiac pathological hypertrophy in rats.MethodsTwenty-nine SD rats were randomly divided into three groups: subcutaneous injection of normal saline combined with intraperitoneal injection of normal saline(Con,n=9) group,subcutaneous injection of normal saline combined with intraperitoneal injection of isoproterenol(ISO,n=10) group and subcutaneous injection of rapamycin(RAP) combined with intraperitoneal injection of ISO(ISO+RAP,n=10) group. After 14-days of continuous treatment,cardiac echocardiogram for left ventricular wall thicknesses and ejection fraction(LVEF) were carried out in Con group(n=9) and ISO group(n=8). Rats were executed by intraperitoneal injection of overdoes 10% chloral hydrate. Heart to body index(HWI) was calculated by heart weight/body weight. Cardiac pathologic alternation was observed by histological sections stained with hematoxylin and eosin(HE) and Masson. The expression of p62 and the ratio of LC3Ⅱ/Ⅰ were measured by Western blot assay. The number of autophagic vacuoles was detected by transmission electron microscopy.ResultsCompared with Con group,the left ventricular wall thickness and LVEF were significantly increased in ISO group(P<0.01). Compared with Con group,the HWI was increased in ISO group and ISO+RAP group(P<0.01). But the HWI was decreased in ISO+RAP group comparison with that of ISO group(P<0.01). Compared with Con group,the alternation of cardiac pathological hypertrophy was marked in ISO group. Moreover,compared with ISO group,cardiac pathological hypertrophy was improved in ISO+RAP group. Compared with Con group,the ratio of LC3Ⅱ/Ⅰwas down-regulated,the protein expression of p62 was up-regulated and the number of autophagic vesicles was significantly reduced in ISO group(P<0.01). However,compared with ISO group,the ratio of LC3Ⅱ/Ⅰwas up-regulated,the protein expression of p62 was down-regulated and the number of autophagic vesicles was significantly increased in ISO + RAP group(P<0.01).ConclusionThe up-regulation of myocardial autophagy can reverse pathological hypertrophy induced by ISO.
引文
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[19]Xue R,Zeng J,Chen Y,et al.Sestrin 1 ameliorates cardiac hypertrophy via autophagy activation[J].J Cell Mol Med,2017,21(6):1193-1205.doi:10.1111/jcmm.13052.
[20]Su M,Chen Z,Wang C,et al.Cardiac-specific overexpression of miR-222 induces heart failure and inhibits autophagy in mice[J].Cell Physiol Biochem,2016,39(4):1503-1511.doi:10.1159/000447853.
[21]Li Z,Song Y,Liu L,et al.miR-199a impairs autophagy and induces cardiac hypertrophy through m TOR activation[J].Cell Death Differ,2017,24(7):1205-1213.doi:10.1038/cdd.2015.95.
[22]Li AL,Lv JB,Gao L.MiR-181a mediates Ang II-induced myocardial hypertrophy by mediating autophagy[J].Eur Rev Med Pharmacol Sci,2017,21(23):5462-5470.doi:10.26355/eurrev_201712_13936.
[23]Huang J,Huang H,Pan W,et al.Renal denervation attenuates cardiac hypertrophy in spontaneously hypertensive rats via regulation of autophagy[J].Mol Med Rep,2017,16(2):2023-2029.doi:10.3892/mmr.2017.6790.
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[2]Tsimploulis A,Sheriff HM,Lam PH,et al.Systolic-diastolic hypertension versus isolated systolic hypertension and incident heart failure in older adults:Insights from the Cardiovascular Health Study[J].Int J Cardiol,2017,235:11-16.doi:10.1016/j.ijcard.2017.02.139.
[3]Bartus K,Podolec J,Sadowski J,et al.Blood pressure reduction in patients with accessory renal arteries and bilateral single renal arteries after catheter-based renal denervation:a prospective study with 3-year follow-up[J].Pol Arch Intern Med,2017,127(6):423-428.doi:10.20452/pamw.4006.
[4]Spiering W,Williams B,Van der Heyden J,et al.Endovascular baroreflex amplification for resistant hypertension:a safety and proof-of-principle clinical study[J].Lancet,2017,390(10113):2655-2661.doi:10.1016/S0140-6736(17)32337-1.
[5]Lobo MD,Ott C,Sobotka PA,et al.Central iliac arteriovenous anastomosis for uncontrolled hypertension:one-year results from the ROX control HTN trial[J].Hypertension,2017,70(6):1099-1105.doi:10.1161/HYPERTENSIONAHA.117.10142.
[6]Li L,Xu J,He L,et al.The role of autophagy in cardiac hypertrophy[J].Acta Biochim Biophys Sin(Shanghai),2016,48(6):491-500.doi:10.1093/abbs/gmw025.
[7]Huang JH,Pan W,Ou DJ,et al.LC3B,a protein that serves as an autophagic marker,modulates angiotensin ii-induced myocardial hypertrophy[J].J Cardiovasc Pharmacol,2015,66(6):576-583.doi:10.1097/FJC.0000000000000306.
[8]Huang JH,Huang CL,Luo YS,et al.Role of MiR-30a in cardiomyocyte autophagy induced by Angiotensin II[J].J Renin Angiotensin Aldosterone Syst,2015,16(1):1-5.doi:10.1177/1470320314562060.
[9]赵美眯,李卓,杨艳,等.注射异丙肾上腺素建立大鼠心肌肥厚模型[J].中国医科大学学报,2017,46(5):406-412.Zhao MM,Li Z,Yang Y,et al.Rat model of isoproterenol‐induced cardiac hypertrophy[J].Journal of China Medical University,2017,46(5):406-412.doi:10.12007/j.issn.0258‐4646.2017.05.006.
[10]Gao L,Guo Y,Liu X,et al.KLF15 protects against isoproterenolinduced cardiac hypertrophy via regulation of cell death and inhibition of Akt/mTOR signaling[J].Biochem Biophys Res Commun,2017,487(1):22-27.doi:10.1016/j.bbrc.2017.03.087.
[11]McMullen JR,Sherwood MC,Tarnavski O,et al.Inhibition of mTOR signaling with rapamycin regresses established cardiac hypertrophy induced by pressure overload[J].Circulation,2004,109(24):3050-3055.doi:10.1161/01.CIR.0000130641.08705.45.
[12]Khalilimeybodi A,Daneshmehr A,Sharif-Kashani B.Investigatingβ-adrenergic-induced cardiac hypertrophy through computational approach:classical and non-classical pathways[J].J Physiol Sci,2018,68(4):503-520.doi:10.1007/s12576-017-0557-5.
[13]Lu J,Sun D,Liu Z,et al.SIRT6 suppresses isoproterenol-induced cardiac hypertrophy through activation of autophagy[J].Transl Res,2016,172:96-112.e6.doi:10.1016/j.trsl.2016.03.002.
[14]Shimizu I,Minamino T.Physiological and pathological cardiac hypertrophy[J].Mol Cell Cardiol,2016,97:245-262.doi:10.1016/j.yjmcc.2016.06.001.
[15]Wang X,Cui T.Autophagy modulation:a potential therapeutic approach in cardiac hypertrophy[J].Am Physiol Heart Circ Physiol,2017,313(2):H304-H319.doi:10.1152/ajpheart.00145.2017.
[16]Lahiri V,Hawkins WD,Klionsky DJ.Watch what you(self-)eat:autophagic mechanisms that modulate metabolism[J].Cell Metab,2019,29(4):803-826.doi:10.1016/j.cmet.2019.03.003.
[17]Papandreou ME,Tavernarakis N.Nucleophagy:from homeostasis to disease[J].Cell Death Differ,2019,26(4):630-639.doi:10.1038/s41418-018-0266-5.
[18]Simonson B,Subramanya V,Chan MC,et al.DDiT4L promotes autophagy and inhibits pathological cardiac hypertrophy in response to stress[J].Sci Signal,2017,10(468).pii:eaaf5967.doi:10.1126/scisignal.aaf5967.
[19]Xue R,Zeng J,Chen Y,et al.Sestrin 1 ameliorates cardiac hypertrophy via autophagy activation[J].J Cell Mol Med,2017,21(6):1193-1205.doi:10.1111/jcmm.13052.
[20]Su M,Chen Z,Wang C,et al.Cardiac-specific overexpression of miR-222 induces heart failure and inhibits autophagy in mice[J].Cell Physiol Biochem,2016,39(4):1503-1511.doi:10.1159/000447853.
[21]Li Z,Song Y,Liu L,et al.miR-199a impairs autophagy and induces cardiac hypertrophy through m TOR activation[J].Cell Death Differ,2017,24(7):1205-1213.doi:10.1038/cdd.2015.95.
[22]Li AL,Lv JB,Gao L.MiR-181a mediates Ang II-induced myocardial hypertrophy by mediating autophagy[J].Eur Rev Med Pharmacol Sci,2017,21(23):5462-5470.doi:10.26355/eurrev_201712_13936.
[23]Huang J,Huang H,Pan W,et al.Renal denervation attenuates cardiac hypertrophy in spontaneously hypertensive rats via regulation of autophagy[J].Mol Med Rep,2017,16(2):2023-2029.doi:10.3892/mmr.2017.6790.
[24]Johnson SC,Yanos ME,Bitto A,et al.Dose-dependent effects of mTOR inhibition on weight and mitochondrial disease in mice[J].Front Genet,2015,6:247.doi:10.3389/fgene.2015.00247.2019-03-122019-04-22