SIRT1通过Wnt/β-catenin通路抑制D-半乳糖诱导的心肌细胞衰老和凋亡  

SIRT1 inhibits D-galactose-induced cardiomyocyte aging and apoptosis through Wnt/β-catenin pathway

作　　者：陈瑞雪 庞淑瑾 陈鑫 郭依宁 方红城 吕洪雪 王陵军[1,2,5] CHEN Ruixue;PANG Shujin;CHEN Xin;GUO Yining;FANG Hongcheng;LÜHongxue;WANG Lingjun(The First Affiliated Hospital of Guangzhou University of Chinese Medicine,Lingnan Medical Research Center,Guangzhou University of Chinese Medicine,Guangzhou 510405,China;Guangdong Clinical Research Institute of Chinese Medicine,Guangzhou 510405,China;Dongguan Hospital,Guangzhou University of Chinese Medicine,Dongguan 523127,China;Shenzhen Hospital of Integrated Traditional Chinese and Western Medicine,Shenzhen 518104,China;State Key Laborato-ry of Traditional Chinese Medicine Syndromes,The First Affiliated Hospital,Guangzhou University of Chinese Medicine,Guangzhou 510405,China)

机构地区：[1]广州中医药大学第一附属医院,广州中医药大学岭南医学研究中心,广东广州510405 [2]广东省中医临床研究院,广东广州510405 [3]广州中医药大学东莞医院,广东东莞523127 [4]深圳市中西医结合医院,广东深圳518104 [5]中医证候全国重点实验室,广东广州510405

出　　处：《中国病理生理杂志》2025年第3期463-471,共9页Chinese Journal of Pathophysiology

基　　金：广东省自然科学基金资助项目(No.2023A1515010282);广东省基础与应用基础研究基金联合基金资助项目(No.2023A1515110747);东莞市社会发展科技项目高水平医院建设专项(No.20231800923372);东莞市社会发展科技项目(No.20211800904402)。

摘　　要：目的:探讨沉默信息调节蛋白1(silent information regulator 1,SIRT1)通过调控Wnt/β-catenin通路对小鼠心肌细胞衰老模型衰老程度及凋亡的影响。方法:采用40μmol/L D-半乳糖(D-galactose,D-Gal)诱导HL-1细胞建立体外衰老模型,SIRT1过表达慢病毒转染HL-1细胞,Western blot验证转染效率。Western blot检测SIRT1、P53、P21、cleaved caspase-3、B细胞淋巴瘤2(B-cell lymphoma-2,Bcl-2)、Bcl-2相关X蛋白(Bcl-2-associated X protein,Bax)、β-catenin、Wnt3a和c-Myc的蛋白表达水平;采用衰老相关β-半乳糖苷酶(senescence-associatedβ-galactosidase,SA-β-Gal)染色检测细胞衰老水平;MTT比色法检测细胞活力;流式细胞术检测细胞凋亡。结果:D-Gal处理小鼠心肌细胞HL-1后衰老相关蛋白P53和P21表达水平增加,SIRT1蛋白表达水平降低,SA-β-Gal染色显示阳性区域增加(P<0.05)。促凋亡蛋白cleaved caspase-3和Bax水平升高,抗凋亡蛋白Bcl-2表达水平降低(P<0.05);细胞活力显著降低(P<0.05);流式细胞术检测细胞凋亡率显著升高(P<0.05),且与D-Gal处理时间呈正相关。过表达SIRT1,经D-Gal处理48 h,HL-1细胞衰老和凋亡水平显著降低(P<0.05)。D-Gal处理HL-1细胞后,β-catenin、c-Myc和Wnt3a蛋白表达水平显著升高,过表达SIRT1后,上述蛋白表达水平显著降低。加入Wnt/β-catenin通路激动剂氯化锂后,与过表达SIRT1并给予D-Gal处理组相比,β-catenin、c-Myc和Wnt3a蛋白表达水平显著升高(P<0.05)。结论:SIRT1通过Wnt/β-catenin通路抑制心肌细胞凋亡并减轻心肌细胞衰老。AIM:To investigate the effect of silent information regulator 1(SIRT1)on the degree of aging and apoptosis in a mouse cardiomyocyte aging model through the regulation of Wnt/β-catenin pathway.METHODS:An in vitro aging model was established by inducing HL-1 cells with 40μmol/L D-galactose(D-Gal).The HL-1 cells were transfected with a lentivirus overexpressing SIRT1,and the transfection efficiency was verified by Western blot.Western blot was used to detect the protein expression levels of SIRT1,P53,P21,cleaved caspase-3,B-cell lymphoma-2(Bcl-2),Bcl-2-associated X protein(Bax),β-catenin,Wnt3a and c-Myc.Senescence-associatedβ-galactosidase(SA-β-Gal)staining was used to detect cellular senescence level.MTT colorimetric assay was used to detect the cell viability,and flow cytometry was used to detect the apoptosis.RESULTS:Treatment of HL-1 mouse cardiomyocytes with D-Gal led to increases in the expression levels of aging-related proteins P53 and P21,as well as an increase in SIRT1 protein level.Additionally,the SA-β-Gal staining showed a significant increase in the positive area(P<0.05).The expression levels of apoptosis-related proteins cleaved caspase-3 and Bax were elevated,while the level of the anti-apoptotic protein Bcl-2 was reduced(P<0.05).There was a marked decrease in cell viability(P<0.05),and flow cytometry analysis demonstrated a significant increase in cell apoptosis rate(P<0.05),which was positively correlated with the duration of D-Gal treatment.Overexpression of SIRT1 notably reduced both aging and apoptosis levels after 48 h of D-Gal treatment(P<0.05).After DGal treatment,the expression levels ofβ-catenin,c-Myc and Wnt3a proteins were up-regulated.However,these levels were reduced when SIRT1 was overexpressed.Moreover,the addition of LiCl,a Wnt/β-catenin pathway agonist,resulted in increased expression levels ofβ-catenin,c-Myc and Wnt3a proteins compared with the group with SIRT1 overexpression and D-Gal treatment(P<0.05).CONCLUSION:SIRT1 inhibits cardiomyocyte apoptosis and alleviates cardi

关 键 词：沉默信息调节蛋白1 心肌细胞 细胞凋亡 细胞衰老 WNT/Β-CATENIN信号通路 

分 类 号：R541[医药卫生—心血管疾病] R363.2[医药卫生—内科学] Q255[医药卫生—临床医学]