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黑色素瘤缺乏因子2在小鼠心肌衰老模型中的表达及意义OACSTPCD

Expression and significance of melanoma deficiency factor 2 in cardiac senescence model

中文摘要英文摘要

背景 心脏衰老是老年人心血管疾病患病率升高的主要原因之一,了解心肌衰老的发病机制以及发现老年心血管疾病的新治疗靶点至关重要.目的 探讨黑色素瘤缺乏因子2(absent in melanoma 2,AIM2)在小鼠心肌衰老模型中的作用.方法 通过生物信息学分析AIM2在年轻小鼠与老龄小鼠心室中具体的表达差异.在动物水平上,将雄性BALB/C小鼠按照年龄分为年轻组(2月龄)和老龄组(14月龄),分别检测两组小鼠心肌中AIM2、凋亡相关斑点样蛋白(apoptosis-associated speck-like protein containing a CARD,ASC)、半胱天冬酶-1(cysteinyl aspartate specific proteinase-1,Caspase-1)在蛋白和 mRNA水平上的变化.在细胞水平上,首先应用不同浓度的D-gal(0 g/L、10 g/L、20 g/L、50 g/L)处理H9C2心肌细胞制备细胞衰老模型,采用CCK-8法检测细胞活力,以确定复制心肌细胞衰老的D-gal浓度;应用选定浓度的D-gal处理H9C2细胞,将细胞分为对照组和衰老组,采用β-半乳糖苷酶染色检测细胞衰老程度,利用Western blot、免疫荧光和RT-qPCR等方法检测心肌细胞中AIM2、ASC、Caspase-1在蛋白和mRNA水平的变化.结果 生物信息学结果显示老龄小鼠心室肌中AIM2表达水平较年轻组升高.动物实验中,老龄组小鼠心室中AIM2、ASC、Caspase-1的蛋白和mRNA水平均较年轻组升高(P<0.05).细胞研究中,使用CCK-8法测定细胞活力,结合细胞状态采用20 g/L的D-gal处理H9C2建立细胞衰老模型,衰老组AIM2、ASC、Caspase-1在蛋白和mRNA水平均较对照组升高(P<0.05).结论 生物信息学、在体与离体实验结果显示,在心肌衰老过程中AIM2在蛋白和mRNA水平均升高,AIM2在心肌衰老的发生和发展中可能发挥了一定作用,为心肌衰老发生机制研究提供了新思路.

Background Cardiac aging is one of the major causes of the increased prevalence of cardiovascular disease in the elderly.Therefore,it is crucial to understand the pathogenesis of cardiac aging and to identify new therapeutic targets for cardiovascular disease in the elderly.Objective To investigate the role of melanoma deficiency factor 2(AIM2)inflammasome in myocardial senescence model.Methods Bioinformatics was used to analyze the difference of AIM2 expression in the ventricles of young and old mice.In animal experiment,male BALB/C mice were divided into young(2 months of age)and old(14 months of age)groups according to age,and AIM2,Apoptosis-associated speck-like protein containing a CARD(ASC),Cysteinyl aspartate specific proteinase-1(Caspase-1)at the protein and mRNA levels were tested in mice of the two groups.At the cellular level,H9C2 cardiomyocytes were treated with different concentrations of(0,10,20,50 g/L)D-gal to prepare cardiomyocyte senescence model,and the cell viability was detected by CCK8 method to determine the optimum D-gal concentration.H9C2 cells were treated with the optimal concentration of D-gal,and the cells were divided into control group and aging group.β-galactosidase staining was used to detect the degree of cell senescence.Western blot,immunofluorescence and RT-qPCR were used to detect the expression of AIM2,ASC and Caspase-1 in cardiomyocytes.Results The results of bioinformatics showed that the expression level of AIM2 in ventricular myocardium of aged mice was higher than that of young mice(P<0.05).In the animal experiment,the protein and mRNA levels of AIM2,ASC,Caspase-1 in the ventricles of the aged group were higher than those of the young group.The cell viability was determined by CCK8 method,and Cardiomyocyte senescence model was established by D-gal treatment of H9C2 with 20g/L.The protein and mRNA levels of AIM2,ASC and Caspase-1 in the aging group were higher than those in the control group(P<0.05).Conclusion The results of bioinformatics,in vivo and in vitro experiments show that the protein and mRNA levels of AIM2 inflammasome increase during myocardial senescence,and AI AIM2 inflammasome may play a certain role in the occurrence and development of myocardial senescence,which provides new ideas for the study of the mechanism of cardiac aging.

王刚;古丽妮尕尔·安外尔;李慧慧;包雅丽;张甜;迪娜·艾尼瓦尔;凌灿;孙湛

新疆医科大学基础医学院病理生理学教研室,新疆乌鲁木齐 830000新疆医科大学基础医学院病理生理学教研室,新疆乌鲁木齐 830000||新疆地方病分子生物学重点实验室,新疆乌鲁木齐 830000

基础医学

心肌衰老AIM2炎性小体生物信息学

cardiac agingAIM2inflammasomebioinformatics

《解放军医学院学报》 2024 (005)

544-550,561 / 8

新疆维吾尔自治区自然科学基金杰出青年科学基金项目(2022D01E53)

10.12435/j.issn.2095-5227.2024.039

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