β1-整合素促进Akt/GSK-3β/β-catenin信号活化并抑制阿霉素诱导的大鼠心肌细胞损伤  

β1-integrin promotes Akt/GSK-3β/β-catenin signaling activation and diminishes cardiac myocytes injury induced by doxorubicin

作　　者：李新[1,2] 刘海涛[1] 陶凌[1] 

机构地区：[1]第四军医大学西京医院心血管内科,陕西省西安市710032 [2]铜川市人民医院超声科

出　　处：《中国心血管病研究》2016年第12期1148-1152,I0012,F0003,共7页Chinese Journal of Cardiovascular Research

摘　　要：目的 研究β1-整合素过表达对阿霉素诱导的大鼠心肌细胞损伤的作用.方法 分离新生SD大鼠心肌细胞,随机分为对照组、空载体组、过表达组、阿霉素组、空载体＋阿霉素组、过表达＋阿霉素组和LY294002预处理组.利用四唑盐比色法（MTT）检测细胞存活,检测心肌细胞乳酸脱氢酶（LDH）释放率、丙二醛（MDA）含量、谷胱甘肽过氧化物酶（GPx）活性、超氧化物歧化酶（SOD）活性,利用实时定量PCR和Western blot检测细胞中β1-整合素以及Akt磷酸化、GSK-3β、β-catenin、Bcl2、Bax的表达,同时利用流式细胞术检测细胞凋亡.结果 空载体组和过表达组转染效率为（68.8±9.5）％和（71.2±8.9）％.与对照组比较,阿霉素组细胞存活率显著降低（P＜0.05）,而过表达＋阿霉素则提高细胞存活率（P＜0.05）.相比于对照组,阿霉素处理促进LDH释放和MDA形成（P＜0.05）,降低GPx和SOD活性（P＜0.05）,过表达＋阿霉素则可以逆转阿霉素作用.阿霉素抑制心肌细胞Akt磷酸化和β-catenin核转移,β1-整合素过表达则促进Akt磷酸化和β-catenin 核转移.阿霉素处理促进细胞凋亡并下调Bcl2表达、上调Bax表达,β1-整合素过表达抑制细胞凋亡并上调Bcl2表达、下调Bax表达.抑制Akt磷酸化显著阻碍β1-整合素的作用.结论 过表达β1-整合素可拮抗阿霉素诱导的细胞毒性,这可能与Akt/GSK-3β/β-catenin信号活化相关.Objective To explore the effect of β1-integrin overexpression on cardiac myocytes injury in- duced by doxorubicin. Methods Cardiac myocytes were prepared from newborn SD rats and randomly allocated into control, blank, overexpression, doxorubicin treatment, blank ＋doxorubicin treatment, overexpression ＋doxoru- bicin treatment, and LY294002 pretreatment group. Cell survival was determined by MTT assay. LDH release, MDA content, GPx activity, and SOD activity were measured using commercial available kits. qRT-PCR and Western blot were used to analyze the expression of β1-integrin, phosphorylated Akt, GSK-3β, β-catenin, Bel2, and Bax. Apoptosis of cardiac myocytes was evaluated by flow cytometry. Results Transfection efficacy were （68.8±9.5）% and （71.2±8.9）% for blank group and overexpression group, respectively. Compared with the con- trol, cell survival was markedly decreased in doxorubiein treatment group（P〈0.05 ）, while β1-integrin overexpres- sion enhanced cell survival （P〈0.05）. Furthermore, doxorubicin promoted LDH release and MDA production as well as reduced GPx and SOD activity （P〈0.05）, while β1-integrin overexpression reversed the effects of doxoru- bicin treatment （P〈0.05）. Doxorubicin restrained Akt phoslphorylation and nuclear translocation of β-catenin in cardiac myocytes, while β1-integrin overexpression enhanced Akt phosphorylation and nuclear translocation of β -catenin. Doxorubicin prompted apoptosis, downregulated Bcl2 expression, and upregulated Bax expression, which were suppressed by β1-integrin overexpression. However, blockade of Akt phosphorylation impeded the ef-feet of β1-integrin overexpression. Conclusion β1-integrin overexpression could counteract cytotoxicity induced by doxorubicin, which might be attributed to activation of Akt/GSK-3β/β-catenin signaling.

关 键 词：Β1-整合素 阿霉素 心肌细胞 AKT磷酸化 

分 类 号：Q95-33[生物学—动物学] R542.2[医药卫生—心血管疾病]