自噬相关基因Rubicon缺失可抑制心脏细胞衰老  

Autophagy-related gene Rubicon deficiency inhibits cellular senescence in the heart

作　　者：何帆 来帅威 张沙沙 董凡 Amber Naz Haniya Mazhar 贺林[1] 朱洪新 He Fan;Lai Shuaiwei;Zhang Shasha;Dong Fan;Amber Naz;Haniya Mazhar;He Lin;Zhu Hongxin(Bio-X Institutes,Key Laboratory for the Genetics of Developmental and Neuropsychiatric Disorders,Ministry of Education,Shanghai Jiao Tong University,Shanghai 200240,China)

机构地区：[1]上海交通大学Bio-X研究院,遗传发育与精神神经疾病教育部重点实验室,上海市200240

出　　处：《中国组织工程研究》2022年第24期3897-3902,共6页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金(81974024),项目负责人:朱洪新;上海市自然科学基金(16ZR1418200),项目负责人:朱洪新。

摘　　要：背景:心脏衰老是心脏疾病的主要危险因素之一,而细胞衰老是心脏衰老的主要机制,在衰老相关心脏疾病中起重要作用。作者前期工作发现自噬相关基因Uvrag缺失促进小鼠心脏细胞衰老,Rubicon是Uvrag的抑制性互作蛋白,其对小鼠心脏细胞衰老的作用尚不明确。目的:探讨Rubicon基因对心脏细胞衰老的影响。方法:使用15月龄野生型小鼠、Rubicon基因缺失小鼠、Uvrag基因缺失小鼠作为观察对象,其中Uvrag基因缺失小鼠作为阳性对照。采用荧光定量PCR检测小鼠心脏组织衰老相关分泌表型相关因子mRNA表达变化,苏木精-伊红染色、天狼猩红染色、衰老相关β-半乳糖苷酶染色进行心肌组织学观察,蛋白免疫印迹检测小鼠心脏组织p53和p16蛋白表达量。实验方案经上海交通大学动物实验伦理委员会批准。结果与结论:①苏木精-伊红染色和天狼猩红染色结果显示,Rubicon基因缺失改善了衰老过程中心肌细胞重塑及心脏纤维化;②衰老相关β-半乳糖苷酶染色发现,Rubicon基因缺失小鼠心脏衰老细胞较野生型对照组显著减少(P<0.05);③荧光定量PCR结果表明,Rubicon基因缺失小鼠心脏白细胞介素1β、白细胞介素6、转化生长因子β和Ⅲ型胶原α1链、基质金属蛋白酶组织抑制因子1 mRNA表达量较野生型对照组显著下调(P<0.05);④免疫印迹结果显示,Rubicon基因缺失导致心脏组织衰老调控关键基因p53表达较野生型对照组显著降低(P<0.05);⑤提示Rubicon基因缺失可显著抑制小鼠心脏细胞衰老并延缓心脏衰老,Rubicon是潜在的抑制心脏细胞衰老、抗心脏衰老及衰老相关心脏疾病的靶分子。BACKGROUND:Cardiac aging is the major risk factor for heart diseases.Cellular senescence is the key mechanism of cardiac aging,which plays an important role in age-related heart disease.Our previous work has shown that deletion of autophagy-related gene Uvrag promotes cellular senescence in the heart.Rubicon is an inhibitory interacting partner of Uvrag.However,the function of Rubicon in cellular senescence in the heart remains unknown.OBJECTIVE:To determine the effect of Rubicon gene on cellular senescence in the heart.METHODS:Wild type and Rubicon-deficient mice at 15 months of age were utilized.Uvrag-deficient mice were applied as observation objects.Uvrag-deficient mice of the same age were used as positive controls.Fluorescence quantitative PCR was used to detect changes in mRNA expression of senescence-related secreted phenotype-related factors in mouse heart tissue.Hematoxylin-eosin staining,Sirius red staining,and senescence-associated β-galactosidase staining were performed to observe myocardial histology.Western blot assay was conducted to measure the expression of p53 and p16 protein in the mouse heart.The protocols were approved by the Animal Expe riment Ethics Committee of Shanghai Jiao Tong Unive rsity.RESULTS AND CONCLUSION:(1) Hematoxylin-eosin staining and Sirius red staining demonstrated that Rubicon deficiency amelio rated myocardial cell remodeling and cardiac fibrosis during aging.(2) Senescence-associated β-galactosidase staining displayed that Rubicon-deficient mice had significantly fewer senescence-associated β-galactosidase-positive cells in hearts from Rubicon-deficient mice than in control mice(P <0.05).(3) Fluorescence quantitative PCR res ults suggested that expression of inte rleukin 1β,inte rleukin 6,transforming growth factor β,type ⅢI collagen α1 chain,and tissue inhibitor of matrix metalloproteinase 1 mRNA in hearts was significantly decreased in Rubicon-deficient mice than that in control mice(P <0.05).(4) Western blot assay res ults showed that p53 protein abundance,a

关 键 词：RUBICON 细胞衰老 心脏衰老 衰老相关分泌表型 P53 小鼠 

分 类 号：R459.9[医药卫生—治疗学] R318[医药卫生—临床医学]