环维黄杨星D对醛固酮诱导型心肌肥厚和凋亡的保护作用及机制  

Protection and mechanism of cyclovirobuxin D on ALD induced cardiac hypertrophy and apoptosis

作　　者：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(School of Pharmacy,Guizhou Medical University&Key Laboratory of Optimal Utilization of Natural Medicine Resources and the High Efficacy Application of Natural Medicinal Resources Engineering Center of Guizhou Province,Guiyang 550025,Guizhou,China)

机构地区：[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,贵州医科大学,贵州贵阳550025

出　　处：《贵州医科大学学报》2024年第2期169-178,共10页Journal of Guizhou Medical University

基　　金：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)。

摘　　要：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 reve

关 键 词：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 

分 类 号：R-332[医药卫生] R285.5R542.2