蒋丹,刘斌,付凌云,徐旖旎,陶玲,沈祥春
JIANG Dan,LIU Bin,FU Linyun,XU Yi'ni,TAO Lin,SHEN Xiangchun

• 1:贵州医科大学药学院&贵州省高等学校天然药物药理与成药性评价重点实验室
• 2:贵州医科大学

摘要(Abstract)：

目的 探讨环维黄杨星D (CVB-D)对醛固酮(ALD)诱导的心肌肥厚和凋亡的保护作用及机制。方法 在C57BL/6小鼠和原代新生大鼠心肌细胞(PNRCMs)中用ALD构建心肌肥厚模型，分为对照组(Control组)、模型组(ALD组)、CVB-D低剂量组(CVB-D.L组)、CVB-D高剂量组(CVB-D.H组)及阳性药螺内酯组(Spir.组);通过超声心动图检测心功能指标左心室射血分数(LVEF)和左心室缩短分数(LVFS),采用HE、Masson和小麦胚芽凝集素荧光(WGA)染色观察小鼠心脏病理变化，MTT检测PNRCMs的细胞活力，吉姆萨染色和乳酸脱氢酶(LDH)检测PNRCMs损伤，cTnT染色观察PNRCMs的表面积，Western blot检测肥厚相关蛋白ANP和BNP、凋亡相关蛋白Bax、Bcl2和Caspase3、Sirt3及PPARγ蛋白的表达水平；使用Sirt3的过表达载体(Ad-Sirt3)进一步分析肥厚ANP、BNP与凋亡相关蛋白Bax、Bcl2及Caspase3的表达。结果 与Control组相比，ALD组小鼠心功能受损，LVEF和LVFS均下降，心脏组织纤维排列紊乱、胶原沉积，心肌细胞活力降低，表面积增大，ANP、BNP、Bax、Caspase3蛋白的表达增加(P<0.05),Sirt3、PPARγ和Bcl2蛋白的表达降低(P<0.05);与ALD组相比，CVB-D显著增强心功能，改善心脏病理变化，提高细胞活力(P<0.05),降低心肌细胞表面积，下调ANP、BNP、Bax、Caspase3蛋白的表达(P<0.05),上调Sirt3、PPARγ和Bcl2蛋白的表达(P<0.05);进一步使用Ad-Sirt3的结果表明Ad-Sirt3能促进CVB-D逆转肥厚和凋亡相关蛋白的表达(P<0.05)。结论 CVB-D减轻ALD诱导的心肌肥厚，抑制心肌细胞凋亡，其机制可能与调节Sirt3/PPARγ信号通路相关。
Objective To investigate the protective effects and mechanisms of cyclovirobuxine D(CVB-D) against cardiac hypertrophy and apoptosis induced by aldosterone(ALD). Methods The cardiac hypertrophy models were replicated by ALD in C57BL/6 mice and primary rat neonatal cardiomyocytes(PNRCMs).Subjects were divided into control group(Control group), model group(ALD group), CVB-D low dose group(CVB-D.L group), CVB-D high dose group(CVB-D.H group), and positive spironolactone group(Spir. group). Cardiac function indicators including LVEF and LVFS were detected by echocardiography. Cardiac pathological changes were observed by HE, Masson and WGA staining. PNRCMs viability was measured by MTT. Giemsa staining and determination of lactate dehydrogenase(LDH) were used to evaluate PNRCMs injury. The surface area of cardiomyocytes was observed by cTnT staining. Western blot was used to detect the expression of hypertrophy-associated proteins ANP and BNP, and apoptosis-associated proteins Bax, Bcl2 and Caspase3, Sirt3, and PPARγ. The overexpression vector of Sirt3(Ad-Sirt3) was further used to analyze the expression of hypertrophy-associated proteins ANP and BNP, and apoptosis-associated proteins Bax, Bcl2, and Caspase3. Results Compared with the control group, mice in the ALD group showed damaged cardiac function indicators, with a decrease in LVEF and LVFS, disturbed fibrous arrangement, collagen deposition, and decreased cell viability(P<0.05), increased cardiomyocyte surface area, increased expression of ANP, BNP, Bax, and Caspase3 proteins(P<0.05), and decreased expression of Sirt3, PPARγ, and Bcl2 proteins(P<0.05). Compared with the ALD group, CVB-D significantly enhanced cardiac function, improved cardiac pathological changes, and increased cell viability(P<0.05), decreased cardiomyocyte surface area, downregulated the expression of ANP, BNP, Bax, and Caspase3 proteins(P<0.05), and upregulated the expression of Sirt3, PPARγ, and Bcl2 proteins(P<0.05). Further studies showed that Ad-Sirt3 promoted the expression of hypertrophy-related and apoptosis-related proteins reversed by CVB-D(P<0.05). Conclusion CVB-D attenuates cardiac hypertrophy and inhibits cardiomyocyte apoptosis induced by ALD, such mechanism might be associated with regulating the Sirt3/PPARγ signalling pathway.

关键词(KeyWords)： 环维黄杨星D;醛固酮;心肌肥厚;细胞凋亡;沉默信息调节因子3;过氧化物酶体增殖物激活受体γ
cycloviroxanthin D;aldosterone;cardiac hypertrophy;apoptosis;sirtuin3;peroxisome proliferators-activated receptor γ

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金(82060729);国家自然科学基金(U1812403-4-4);; 贵州省科技厅科技计划项目(黔科合中引地[2023]003)

作者(Author): 蒋丹,刘斌,付凌云,徐旖旎,陶玲,沈祥春
JIANG Dan,LIU Bin,FU Linyun,XU Yi'ni,TAO Lin,SHEN Xiangchun

DOI: 10.19367/j.cnki.2096-8388.2024.02.002

参考文献(References)：

• [1] 《中国心血管健康与疾病报告 2021》编写组.《中国心血管健康与疾病报告 2021》要点解读[J].中国心血管杂志，2022,27(4):305-318.
• [2] NAKAMURA K,MIYOSHII T,YOSHIDA M,et al.Pathophysiology and treatment of diabetic cardiomyopathy and heart failure in patients with diabetes mellitus[J].Int J Mol Sci,2022,23(7):3587.
• [3] THAM Y K,BERNARDO B C,OOI J Y,et al.Pathophysiology of cardiac hypertrophy and heart failure:signaling pathways and novel therapeutic targets[J].Arch Toxicol,2015,89(9):1401-1438.
• [4] GUITART-MAMPEL M,URQUIZA P,BORGES J I,et al.Impact of aldosterone on the failing myocardium:insights from mitochondria and adrenergic receptors signaling and function[J].Cells,2021,10(6):1552.
• [5] JIA G,AROOR A R,JIA C,et al.Endothelial cell senescence in aging-related vascular dysfunction[J].Biochim Biophys Acta Mol Basis Dis,2019,1865:1802-1809.
• [6] SUNDARESAN N R,GUPTA M,KIM G,et al.Sirt3 blocks the cardiac hypertrophic response by augmenting Foxo3a-dependent antioxidant defense mechanisms in mice[J].J Clin Invest,2009,119(9):2758-2771.
• [7] LIU C,XIONG Q,LI Q,et al.CHD7 regulates bone-fat balance by suppressing PPAR-γ signaling[J].Nat Commun,2022,13(1):1989.
• [8] TSUTSUI H,HAYASHIDANI S,SUEMATSU N,et al.Pioglitazone,a peroxisome proliferator-activated receptor-gamma agonist,attenuates left ventricular remodeling and failure after experimental myocardial infarction[J].Circulation,2002,106(24):3126-3132.
• [9] GUO X,YAN F,SHAN X,et al.SIRT3 inhibits Ang Ⅱ-induced transdifferentiation of cardiac fibroblasts through β-catenin/PPAR-γ signaling[J].Life Sci,2017,186:111-117.
• [10]赵静，赵颖，李晗.大蒜素通过调控SIRT3/PPARγ信号通路促进心衰大鼠血管再生和心功能改善[J].中国循证心血管医学杂志，2021,13(8):1011-1014.
• [11]ZHOU L,TANG H,WANG F,et al.Cyclovirobuxine D inhibits cell proliferation and migration and induces apoptosis in human glioblastoma multiforme and lowgrade glioma[J].Oncol Rep,2020,43(3):807-816.
• [12]GUO Q,GUO J,YANG R,et al.Cyclovirobuxine D attenuates doxorubicin-Induced cardiomyopathy by suppression of oxidative damage and mitochondrial biogenesis impairment[J].Oxid Med Cell Longev,2015,2015:151972.
• [13]张光琼，王声全，付凌云，等.环维黄杨星D调控Ca2+/CaMK Ⅱ信号改善醛固酮诱导心肌细胞损伤的实验研究[J].中国药理学通报，2021,37(4):564-570.
• [14]陈世平，徐旖旎，杨宇，等.环维黄杨星D对醛固酮联合高盐诱导大鼠实验性心衰模型心功能障碍的影响[J].中药材，2019,42(8):1885-1889.
• [15]MONTAIGNE D,BUTRUILLE L,STAELS B.PPAR control of metabolism and cardiovascular functions[J].Nat Rev Cardiol,2021,18(12):809-823.
• [16]FERREIRA N S,TOSTES R C,PARADIS P,et al.Aldosterone,inflammation,immune system,and hypertension[J].Am J Hypertens,2021,34(1):15-27.
• [17]BUFFOLO F,TETTI M,MULATERO P,et al.Aldosterone as a mediator of cardiovascular damage[J].Hypertension,2022,79(9):1899-1911.
• [18]SU D,GONG Y,LI S,et al.Cyclovirobuxine D,a cardiovascular drug from traditional Chinese medicine,alleviates inflammatory and neuropathic pain mainly via inhibition of voltage-gated Ca3.2 channels[J].Front Pharmacol,2022,13:1081697.
• [19]JIANG Z,FU L,XU Y,et al.Cyclovirobuxine D protects against diabetic cardiomyopathy by activating Nrf2-mediated antioxidant responses[J].Sci Rep,2020,10(1):6427.
• [20]GAO G,FU L,XU Y,et al.Cyclovirobuxine D ameliorates experimental diabetic cardiomyopathy by inhibiting cardiomyocyte pyroptosis via NLRP3 in vivo and in vitro[J].Front Pharmacol,2022,13:906548.
• [21]DEI RE D P,AMGALAN D,LINKERMANN A,et al.Fundamental mechanisms of regulated cell death and implications for heart disease[J].Physiol Rev,2019,99(4):1765-1817.
• [22]CHEN J,CHEN S,ZHANG B,et al.SIRT3 as a potential therapeutic target for heart failure[J].Pharmacol Res,2021,165:105432.
• [23]TOMCZYK M M,CHEUNG K G,XIANG B,et al.Mitochondrial sirtuin-3 (sirt3) prevents doxorubicin-induced dilated cardiomyopathy by modulating protein acetylation and oxidative stress[J].Circ Heart Fail,2022,15(5):e008547.
• [24]SONG S,DING Y,DAI G L,et al.Sirtuin 3 deficiency exacerbates diabetic cardiomyopathy via necroptosis enhancement and NLRP3 activation[J].Acta Pharmacol Sin,2021,42(2):230-241.
• [25]LEGCHENKO E,CHOUVARINE P,BORCHERT P,et al.PPARγ agonist pioglitazone reverses pulmonary hypertension and prevents right heart failure via fatty acid oxidation[J].Sci Transl Med,2018,10(438):0303.
• [26]XIONG S P,SUN H J,CAO X,et al.Polysulfide protects against diabetic cardiomyopathy through sulfhydration of peroxisome proliferator-activated receptor-γ and sirtuin 3[J].Antioxid Redox Signal,2023,38(1-3):1-17.

文章评论(Comment)：