LncRNA LOC105378097 inhibits cardiac mitophagy in natural ageing mice.
LncRNA LOC105378097 inhibits cardiac mitophagy in natural ageing mice.
复制标题
LncRNA LOC105378097 抑制自然衰老小鼠的心脏线粒体自噬
DOI：
10.1002/ctm2.908
复制
发表时间：
2022-06
影响因子：
10.6
通讯作者：
中科院分区：
文献类型：
--
作者：
心脏衰老的发展是老年人慢性残疾、疾病和死亡的主要原因。充足的证据已经证实，线粒体自噬在衰老心脏中显着减少具有关键作用。然而，衰老心脏线粒体自噬缺陷的潜在机制却鲜为人知。本研究旨在探讨lncRNA LOC105378097（衰老-线粒体自噬相关LncRNA，lncR-SMAL）在心脏衰老过程中对线粒体自噬作用的潜在机制。通过实时定量聚合酶链反应测量不同年龄人类血清和不同年龄小鼠心脏中lncR-SMAL的表达。通过超声心动图和压力-容积测量系统评估lncR-SMAL对小鼠心功能的影响。通过苏木精-伊红染色、衰老相关 β-半乳糖苷酶染色、流式细胞术和衰老相关标记 p53 和 p21 表达的蛋白质印迹分析来评估心脏衰老。通过蛋白质印迹、mRFP-GFP-LC3 腺病毒颗粒转染和 mito-Keima 染色评估心肌细胞线粒体自噬。通过分子对接、RNA 免疫沉淀 (RIP) 和 RNA Pull-down 测定验证了 lncR-SMAL 和 Parkin 之间的相互作用。进行泛素化检测以探讨 Parkin 抑制的分子机制。通过电镜检查、JC-1染色和耗氧率分析研究lncR-SMAL对线粒体功能的影响。心脏富集的lncR-SMAL在人的血清中在60岁时达到表达峰值。外源性lncRNA SMAL的过度表达与自然衰老完全一样，会导致心脏功能恶化，并通过降低Parkin蛋白水平来抑制相关的心肌细胞线粒体自噬。在小鼠中，lncR-SMAL 可逆转 Parkin 过度表达引起的心脏衰老和线粒体自噬的改善。相比之下，AC16细胞中lncR-SMAL的缺失导致Parkin蛋白上调，改善线粒体自噬和线粒体功能障碍，从而减轻心脏衰老。此外，我们通过计算对接分析、pull-down和RIP实验发现了lncR-SMAL和Parkin蛋白之间的相互作用。这将有助于 lncR-SMA 对 Parkin 泛素化的促进作用并降低 Parkin 蛋白的稳定性。本研究首次证明了一种富含心脏的lncRNA SMAL，它可以通过下调Parkin蛋白来抑制心肌细胞的线粒体自噬，从而进一步导致自然衰老小鼠的心脏衰老和心功能障碍。 (1) 与年轻对照相比，60 岁以上人类受试者的血清样本中以及衰老心肌细胞的细胞核和细胞质中，心脏富集的 lncR-SMAL 水平显着升高。 (2) lncR-SMAL 的人工过度表达通过抑制 Parkin 介导的线粒体自噬诱导心脏衰老。心肌细胞中内源性 lncR-SMAL 的敲低部分消除了细胞衰老。 (3)LncR-SMAL与Parkin蛋白相互作用并促进衰老心肌细胞中Parkin的泛素蛋白酶体降解。
The development of heart ageing is the main cause of chronic disability, disease and death in the elderly. Ample evidence has established a pivotal role for significantly reduced mitophagy in the ageing heart. However, the underlying mechanisms of mitophagy deficiency in ageing heart are little known. The present study aimed to explore the underlying mechanisms of lncRNA LOC105378097 (Senescence‐Mitophagy Associated LncRNA, lncR‐SMAL) actions on mitophagy in the setting of heart ageing. The expression of lncR‐SMAL was measured in serum from different ages of human and heart from different ages of mice through a quantitative real‐time polymerase chain reaction. The effects of lncR‐SMAL on heart function of mice were assessed by echocardiography and pressure‐volume measurements system. Cardiac senescence was evaluated by hematoxylin‐eosin staining, senescence‐associated β‐galactosidase staining, flow cytometry and western blot analysis of expression of ageing related markes p53 and p21. Cardiomyocyte mitophagy was assessed by western blot, mRFP‐GFP‐LC3 adenovirus particles transfection and mito‐Keima staining. Interaction between lncR‐SMAL and Parkin was validated through molecular docking, RNA immunoprecipitation (RIP) and RNA pull‐down assay. Ubiquitination assay was performed to explore the molecular mechanism of Parkin inhibition. The effects of lncR‐SMAL on mitochondrial function were investigated through electron microscopic examination, JC‐1 staining and oxygen consumption rates analysis. The heart‐enriched lncR‐SMAL reached the expression crest in the serum of human at an age of 60. Exogenously overexpression of lncRNA SMAL deteriorated cardiac function exactly as natural ageing and inhibited the associated cardiomyocytes mitophagy by depressing Parkin protein level. Improved heart ageing and mitophagy caused by Parkin overexpression were reversed by lncR‐SMAL in mice. In contrast, the loss of lncR‐SMAL in AC16 cells induced the upregulation of Parkin protein and ameliorated mitophagy and mitochondrial dysfunction, resulting in alleviated cardiac senescence. Besides, we found the interaction between lncR‐SMAL and Parkin protein through computational docking analysis, pull‐down and RIP assay. This would contribute to the promotive effect of lncR‐SMAL on Parkin ubiquitination and decrease Parkin protein stability. The present study for the first time demonstrates a heart‐enriched lncRNA, SMAL, that inhibits the mitophagy of cardiomyocytes via the downregulation of Parkin protein, which further contributes to heart ageing and cardiac dysfunction in natural ageing mice. (1) The heart‐enriched lncR‐SMAL is robustly increased in its level in serum samples of human subjects over 60‐year old relative to young controls, and in both nucleus and cytoplasm of senescent cardiomyocytes. (2) Artificial overexpression of lncR‐SMAL induces cardiac senescence by inhibiting Parkin‐mediated mitophagy. Knockdown of endogenous lncR‐SMAL in cardiomyocytes partially abrogated cell senescence. (3) LncR‐SMAL interacts with Parkin protein and promotes ubiquitin‐proteasome degradation of Parkin in senescent cardiomyocytes.
登录
查看更多内容
影响因子：
20.1
作者：
Chen K;Wang S;Sun QW;Zhang B;Ullah M;Sun Z
通讯作者：
Sun Z
DOI：
10.15252/embj.201592862
发表时间：
2016-04-01
期刊：
The EMBO journal
影响因子：
--
作者：
Correia-Melo C;Marques FD;Anderson R;Hewitt G;Hewitt R;Cole J;Carroll BM;Miwa S;Birch J;Merz A;Rushton MD;Charles M;Jurk D;Tait SW;Czapiewski R;Greaves L;Nelson G;Bohlooly-Y M;Rodriguez-Cuenca S;Vidal-Puig A;Mann D;Saretzki G;Quarato G;Green DR;Adams PD;von Zglinicki T;Korolchuk VI;Passos JF
通讯作者：
Passos JF
lncRNA OIP5-AS1/Cyrano海绵RNA结合蛋白HUR。
影响因子：
14.9
作者：
Kim J;Abdelmohsen K;Yang X;De S;Grammatikakis I;Noh JH;Gorospe M
通讯作者：
Gorospe M
影响因子：
11.4
作者：
Jin Q;Li R;Hu N;Xin T;Zhu P;Hu S;Ma S;Zhu H;Ren J;Zhou H
通讯作者：
Zhou H
影响因子：
--
作者：
Ikeda Y;Sciarretta S;Nagarajan N;Rubattu S;Volpe M;Frati G;Sadoshima J
通讯作者：
Sadoshima J