安石榴苷对阿霉素所致心脏毒性的改善作用及其可能机制

Improvement of punicalagin on adriamycin-induced cardiotoxicity and its possible mechanism

目的探讨安石榴苷对阿霉素所致心脏毒性的改善作用,并基于5′-单磷酸腺苷酸活化蛋白激酶(AMPK)/核因子红细胞2相关因子2(Nrf2)通路及细胞凋亡、氧化应激探讨其可能的作用机制。方法将48只雄性C57BL/6小鼠随机分为对照组、阿霉素组、阿霉素+安石榴苷组、阿霉素+安石榴苷+Compound c组,每组12只。除对照组外,给予其他各组小鼠一次性腹腔注射阿霉素。同时,给予阿霉素+安石榴苷组小鼠腹腔注射安石榴苷,给予阿霉素+安石榴苷+Compound c组小鼠腹腔注射安石榴苷后再腹腔注射Compound c,给予对照组小鼠腹腔注射等量生理盐水。干预7 d后,观察各组小鼠心脏组织形态结构,检测各组小鼠心脏功能[左室压力上升最大速率(+dp/dt_(max))、左室压力下降最大速率(-dp/dt_(max))],血清肌钙蛋白T(cTnT)水平,以及心脏组织活性氧簇(ROS)水平和cleaved Caspase-3、细胞色素c(Cyt-c)、gp91^(phox)、超氧化物歧化酶2(SOD2)、磷酸化AMPK(p-AMPK)、AMPK、Nrf2和血红素加氧酶1(HO-1)蛋白表达水平。结果与对照组比较,阿霉素组小鼠的+dp/dt_(max)、-dp/dt_(max)降低(P<0.05),心肌纤维结构紊乱,出现胞质空泡化,血清cTnT水平升高,心脏组织ROS水平及cleaved Caspase-3、Cyt-c、gp91^(phox)蛋白表达水平上调,SOD2、p-AMPK、Nrf2和HO-1蛋白表达水平下调(均P<0.05);与阿霉素组比较,阿霉素+安石榴苷组小鼠的+dp/dt_(max)、-dp/dt_(max)升高(P<0.05),心肌纤维结构改善,胞质空泡化减少,血清cTnT水平降低,心脏组织ROS水平及cleaved Caspase-3、Cyt-c、gp91^(phox)蛋白表达水平下调,SOD2、p-AMPK、Nrf2和HO-1蛋白表达水平上调(均P<0.05);与阿霉素+安石榴苷组比较,阿霉素+安石榴苷+Compound c组小鼠的+dp/dt_(max)、-dp/dt_(max)降低(P<0.05),心肌纤维结构紊乱,胞质空泡化加重,血清cTnT水平升高,心脏组织ROS水平及cleaved Caspase-3、Cyt-c、gp91^(phox)蛋白表达水平上调,SOD2、p-AMPK、Nrf2和HO-1蛋白表达水平下调(均P<0.05)。4组小鼠AMPK蛋白表达水平差异无统计学意义(P>0.05)。结论安石榴苷可能通过AMPK/Nrf2通路抑制细胞凋亡和氧化应激损伤,从而减轻阿霉素引起的心脏毒性。

Objective To investigate the improvement of punicalagin on adriamycin-induced cardiotoxicity,and to explore its possible mechanism based on adenosine 5′-monophosphate activated protein kinase(AMPK)/nuclear factor erythrocyte 2-related factor 2(Nrf2)pathway,and cell apoptosis and oxidative stress.Methods A total of 48 male C57BL/6 mice were randomly divided into control group,adriamycin group,adriamycin+punicalagin group,or adriamycin+punicalagin+Compound c group,with 12 mice in each group.Except for the control group,mice in the remaining groups received one-time intraperitoneal injection of adriamycin;simultaneously,the adriamycin+punicalagin group received intraperitoneal injection of punicalagin,and the adriamycin+punicalagin+Compound c group received intraperitoneal injection of punicalagin,and then received intraperitoneal injection of Compound c,and the control group received intraperitoneal injection of normal saline with equivalent volume.After 7 days of intervention,cardiac morphological structures of various groups were observed,and cardiac function(maximum rate of left ventricular pressure rise[+dp/dt_(max)]and_(max)imum rate of left ventricular pressure drop[-dp/dt_(max)]),and the levels of serum cardiac troponin T(cTnT),reactive oxygen species(ROS)in cardiac tissues,as well as the expressions of cleaved Caspase-3,cytochrome c(Cyt-c),gp91^(phox),superoxide dismutase 2(SOD2),phosphorylated AMPK(p-AMPK),AMPK,Nrf2 and heme oxygenase 1(HO-1)proteins were detected in various groups.Results Compared with the control group,the adriamycin group yielded decreased+dp/dt_(max)and-dp/dt_(max)(P<0.05),presented as disordered myocardial fibers structure and cytoplasmic vacuolation,and exhibited elevated levels of serum cTnT and ROS in cardiac tissues,as well as up-regulated expressions of cleaved Caspase-3,Cyt-c and gp91^(phox)proteins,while down-regulated expressions of SOD2,p-AMPK,Nrf2 and HO-1 proteins in cardiac tissues(all P<0.05).Compared with the adriamycin group,the adriamycin+punicalagin group exhibited elevated+dp/dt_(max)and-dp/dt_(max)(P<0.05),presented as improved myocardial fibers structure and decreased cytoplasmic vacuolation,and interpreted decreased levels of serum cTnT and ROS in cardiac tissues,as well as down-regulated expressions of cleaved Caspase-3,Cyt-c and gp91^(phox)proteins,while up-regulated expressions of SOD2,p-AMPK,Nrf2 and HO-1 proteins in cardiac tissues(all P<0.05).Compared with the adriamycin+punicalagin group,the adriamycin+punicalagin+Compound c group interpreted decreased+dp/dt_(max)and-dp/dt_(max)(P<0.05),presented as disordered myocardial fibers structure and exacerbation of cytoplasmic vacuolation,and depicted elevated levels of serum cTnT and ROS in cardiac tissues,as well as up-regulated expressions of cleaved Caspase-3,Cyt-c and gp91^(phox)proteins,while down-regulated expressions of SOD2,p-AMPK,Nrf2 and HO-1 proteins in cardiac tissues(all P<0.05).There was no statistically significant difference in the expression of AMPK protein between the four groups(P>0.05).Conclusion Punicalagin may relieve adriamycin-induced cardiotoxicity through inhibiting cell apoptosis and oxidative stress injury via AMPK/Nrf2 pathway.