摘要
目的 阿霉素(doxorubicin, Dox)的临床应用受到其剂量依赖的心脏毒性的限制,本研究拟探究槲皮素(Quercetin, Que)在Dox诱导的细胞毒性中的作用机制。方法 将小鼠随机分为Con组、Que处理组、Dox组、Dox-Que组,将分离的乳鼠心肌原代细胞(neonatal mouse cardiomyocytes, nmCMs)随机分为Dox-Vehicle组、Dox-Que组、Dox-Que-SB组、Dox-SB组及Dox-3 MA组。细胞活力以CCK-8试剂盒检测、同时检测ATP生成及ROS生成,此外以Western blot方法检测线粒体自噬标志蛋白的表达情况。结果 在体研究证实,Que可通过抑制线粒体自噬并降低心肌纤维化抵抗Dox诱导的心肌毒性。体外研究表明,Dox处理可显著降低nmCMs细胞活性,ATP含量及增加ROS生成,同时线粒体自噬被显著激活(P<0.05),Que预处理可通过抑制线粒体自噬的激活而缓解Dox诱导的心肌细胞损伤,并降低ROS生成及增加线粒体ATP含量。我们进一步的研究证实Que对线粒体自噬的抑制作用是Pink1/Parkin分子通路介导的,而SB处理可通过激活线粒体自噬而部分抵消Que对Dox诱导的心肌毒性的保护作用。结论 Que通过抑制Pink1/Parkin介导的线粒体自噬的激活,促进ATP生成及降低ROS活性,缓解Dox引起的心肌毒性。
Objective The clinical application of doxorubicin(Dox) is limited by its dose-dependent cardiotoxicity. This study was established to elucidate the mechanism by which Quercetin(Que) acts in Dox-induced cytotoxicity. Methods Mice were randomly divided into a Con group, Que group, Dox group and Dox-Que group. Neonatal mouse cardiomyocytes were isolated and randomly divided into a Dox-Vehicle group, Dox-Que group, Dox-Que-SB group, Dox-SB group and Dox-3 MA group. Cell viability was detected by CCK-8 kit, while ATP content and ROS production were determined simultaneously. In addition, Western blot was used to determine the expression of mitophagy marker proteins. Results In vivo studies confirmed that Que can reduce Dox-induced cardiotoxicity by inhibiting mitophagy and reducing myocardial fibrosis. In vitro studies showed that Dox treatment significantly reduced the cell activity of neonatal mouse cardiomyocytes and ATP content, increased ROS production, and significantly activated mitophagy(P<0.05). Que pretreatment alleviated Dox-induced cardiomyocyte injury by inhibiting the activation of mitophagy while reducing ROS production and increasing mitochondrial ATP content. Our study further confirmed that the inhibitory effect of Que on mitophagy was mediated by the Pink1/Parkin-dependent signaling pathway, and that SB treatment partially offset the protective effect of Que on Dox-induced cardiotoxicity by activating mitophagy. Conclusions Que can inhibit Pink1/Parkin-mediated mitophagy activation, promote ATP production, reduce ROS production and alleviate Dox-induced cardiotoxicity.
作者
郭红
王少玮
谢文静
周俊
蒋书云
刘金成
杨剑
金屏
GUO Hong;WANG Shaowei;XIE Wenjing;ZHOU Jun;JIANG Shuyun;LIU Jincheng;YANG Jian;JIN Ping(Department of Cardiovascular Surgery,Xijing Hospital,Air Force Medical University,Xi’an 710032,China)
出处
《中国比较医学杂志》
CAS
北大核心
2022年第7期1-9,共9页
Chinese Journal of Comparative Medicine
基金
国家自然科学基金(81870216)。
作者简介
郭红(1983-),女,研究方向:心肌保护。E-mail:guo_hongxj@126.com;通信作者:金屏(1984-),男,副主任医师,硕士,研究方向:心肌保护。E-mail:bohua001@126.com。
