Sun Zhen, Li Lihua, Wu Yao, Zhang Lili, Zang Guangyao, Qian Yongjiang, Yao Haipeng, Mao Xiang, Wang Zhongqun
Department of Cardiology, Affiliated Hospital of Jiangsu University, Zhenjiang 212001, China.
Institute of Cardiovascular Diseases, Jiangsu University, Zhenjiang 212001, China.
Cardiovasc Res. 2025 Apr 22;121(2):296-310. doi: 10.1093/cvr/cvae263.
Microcalcification increases the vulnerability of plaques and has become an important driver of acute cardiovascular events in diabetic patients. However, the regulatory mechanisms remain unclear. DJ-1, a multifunctional protein, may play a potential role in the development of diabetic complications. Therefore, this study aims to explore the relationship between DJ-1 and microcalcification in diabetic plaques and investigate the mechanisms.
The regulatory relationship between DJ-1 and diabetic vascular microcalcification was determined in anterior tibial arteries from diabetic foot amputated patients, a diabetic apolipoprotein E-deficient (ApoE-/-) mouse model, and a vascular smooth muscle cell (VSMC) model. The ubiquitination and acetylation levels of DJ-1 were detected, and the acetylation-ubiquitination crosstalk was explored. Then, the regulatory effects of DJ-1 on receptor for advanced glycation end products (RAGE) were clarified. Further, the role of DJ-1 in collagen-matrix vesicles (MVs) interaction in diabetic microenvironment was observed. The collagen interacting surface protein of MVs was verified with proteomics and the biomimetic MVs model. In clinical samples, the number of microcalcification nodules in anterior tibial artery plaques was negatively correlated with DJ-1 expression. In diabetic ApoE-/- mice and VSMCs models, knocking down DJ-1 significantly increased the number of microcalcified nodules. N-acetyltransferase 10 (NAT10) was an acetyltransferase of DJ-1. NAT10 could crosstalk the ubiquitination of DJ-1 and enhance the ubiquitination of DJ-1 by E3 ubiquitin ligase tripartite motif-containing protein 32 (TRIM32). Besides, the knockdown of DJ-1 activated signal transducer and activator of transcription 1 (STAT1), and then STAT1 could bind to RAGE promoter, thus up-regulating RAGE. Furthermore, the knockdown of DJ-1 significantly promoted collagen-MVs interaction in diabetic microenvironment. Milk fat globule epidermal growth factor 8 (MFGE8) may serve as a collagen-interacting protein. The coating of MFGE8 protein could increase the interaction between collagen and biomimetic MVs.
In the diabetic microenvironment, DJ-1 was a protective factor for vascular microcalcification. NAT10- and TRIM32-mediated acetylation-ubiquitination crosstalk resulted in the degradation of DJ-1. The decrease of DJ-1 could activate DJ-1/STAT1/RAGE microcalcification signal. Further, under the stimulation of DJ-1-mediated microcalcification signal, VSMCs released MVs with high abundance of MFGE8. MFGE8 promoted collagen-MVs interaction and finally accelerated the formation of microcalcification.
微钙化增加了斑块的易损性，并已成为糖尿病患者急性心血管事件的重要驱动因素。然而，其调控机制仍不清楚。DJ-1是一种多功能蛋白，可能在糖尿病并发症的发生发展中发挥潜在作用。因此，本研究旨在探讨DJ-1与糖尿病斑块中微钙化之间的关系，并研究其机制。
在糖尿病足截肢患者的胫前动脉、糖尿病载脂蛋白E缺陷（ApoE-/-）小鼠模型和血管平滑肌细胞（VSMC）模型中确定DJ-1与糖尿病血管微钙化之间的调控关系。检测DJ-1的泛素化和乙酰化水平，并探讨乙酰化-泛素化的相互作用。然后，阐明DJ-1对晚期糖基化终末产物受体（RAGE）的调控作用。此外，观察DJ-1在糖尿病微环境中胶原-基质小泡（MVs）相互作用中的作用。用蛋白质组学和仿生MVs模型验证MVs的胶原相互作用表面蛋白。在临床样本中，胫前动脉斑块中微钙化结节的数量与DJ-1表达呈负相关。在糖尿病ApoE-/-小鼠和VSMCs模型中，敲低DJ-1显著增加了微钙化结节的数量。N-乙酰转移酶10（NAT10）是DJ-1的乙酰转移酶。NAT10可以与DJ-1的泛素化相互作用，并通过E3泛素连接酶含三联基序蛋白32（TRIM32）增强DJ-1的泛素化。此外，敲低DJ-1激活了信号转导子和转录激活子1（STAT1），然后STAT1可以与RAGE启动子结合，从而上调RAGE。此外，敲低DJ-1显著促进了糖尿病微环境中胶原-MVs的相互作用。乳脂肪球表皮生长因子8（MFGE8）可能作为一种胶原相互作用蛋白。MFGE8蛋白的包被可以增加胶原与仿生MVs之间的相互作用。
在糖尿病微环境中，DJ-1是血管微钙化的保护因子。NAT10和TRIM32介导的乙酰化-泛素化相互作用导致DJ-1的降解。DJ-1的减少可激活DJ-1/STAT1/RAGE微钙化信号。此外，在DJ-1介导的微钙化信号刺激下，VSMCs释放富含MFGE8的MVs。MFGE8促进胶原-MVs相互作用，最终加速微钙化的形成。
