C/EBPα-Mediated Transcriptional Activation of PIK3C2A Regulates Autophagy, Matrix Metalloproteinase Expression, and Phenotypic of Vascular Smooth Muscle Cells in Aortic Dissection

• Published in: Journal of Immunology Research Vol. 2022; pp. 1 - 14
• Main Authors: Lu, Heng, Chen, Yi, Chen, Yinhai, Huang, Lingchen, Chen, Liangwan
• Format: Journal Article
• Language: English
• Published: New York Hindawi 12.09.2022; John Wiley & Sons, Inc; Hindawi Limited
• Subjects: Advertising executives; Antibodies; Aorta; Aortic dissection; Autophagy; Dissecting aneurysm; Dissection; Extracellular matrix; Gelatinase A; Gelatinase B; Genetic aspects; Genetic transcription; Genotype & phenotype; Homeostasis; Immunohistochemistry; Immunology; Kinases; Laboratory animals; Major histocompatibility complex; Matrix metalloproteinase; Metalloproteinase; Muscle contraction; Phenotypes; Proteins; Rapamycin; Rodents; siRNA; Smooth muscle; Surgery; Transcription activation; Transfection; China; United States > US
• ISSN: 2314-8861; 2314-7156
• DOI: 10.1155/2022/7465353

Purpose. To investigate the function of C/EBPα in the development of aortic dissection (AD) and the underlying mechanism. Methods. Aortic vascular smooth muscle cells (VSMCs) were isolated, cultured, and identified from AD rats. Then, C/EBPα and PIK3C2A were knockdown or overexpressed by siRNA or plasmid transfection, respectively. Rapamycin or 3-MA was utilized to stimulate and restrain autophagy of VSMCs, respectively. Western blot was used to evaluate the expression levels of C/EBPα, PIK3C2A, LC3, Beclin-1, p62, MMP-2, MMP-9, α-SMA, SM-MHC, and OPN. The pathological status of aortic ring was evaluated by stretch stress, and ChIP assay was used to analyze the binding between C/EBPα and PIK3C2A. C/EBPα shRNA was injected into tail vein to observe the effect of C/EBPα knockdown in vivo on phenotype, autophagy of aortic vascular tissue by immunohistochemical staining and Western blot. Results. The protein levels of C/EBPα, PIK3C2A, MMP-2, MMP-9, and LC3 in the aorta of AD rats were all upregulated significantly. C/EBPα and rapamycin promoted notable upregulation of the synthesized proteins (OPN), PIK3C2A, matrix metalloproteinases, LC3, and Beclin-1 in VSMCs, while suppressed contractile proteins (α-SMA and SM-MHC) and p62. The opposite results were observed in the C/EBPα-knockdown VSMCs, PIK3C2A-knockdown VSMCs, or VSMCs treated with 3-MA. C/EBPα, PIK3C2A, and LC3 were dramatically upregulated by the stimulation of 3 g and 5 g stretch stress. The downregulated contractile proteins, upregulated synthetic proteins, activated autophagy, and aggravated pathological state in 5 g stretch stress-treated aortic rings were significantly reversed by the knockdown of C/EBPα. ChIP results indicated that there was a binding site for C/EBPα in the promoter of PIK3C2A. C/EBPα also downregulated α-SMA level and upregulated OPN levels in AD rats in vivo. Conclusion. Our data indicated that during the development of AD, C/EBPα regulated the transition of VSMC phenotype and extracellular matrix remodeling by activating autophagy through regulating the transcriptional activity of PIK3C2A promoter.