Abstract
Background Pathological changes of the adrenal gland and the possible underlying molecular mechanisms are currently unclear in the case of atherosclerosis (AS) combined with chronic stress (CS). Methods New Zealand white rabbits were used to construct a CS and AS animal model. Proteomics and bioinformatics were employed to identify hub proteins in the adrenal gland related to CS and AS. Hub proteins were detected using immunohistochemistry, immunofluorescence assays, and Western blotting. Real-time quantitative polymerase chain reaction (RT-qPCR) was used to analyze the expression of genes. In addition, a neural network model was constructed. The quantitative relationships were inferred by cubic spline interpolation. Enzymatic activity of mitochondrial citrate synthase and OGDH was detected by the enzymatic assay kit. Function of citrate synthase and OGDH with knockdown experiments in the adrenal cell lines was performed. Furthermore, target genes-TF-miRNA regulatory network was constructed. Coimmunoprecipitation (IP) assay and molecular docking study were used to detect the interaction between citrate synthase and OGDH. Results Two most significant hub proteins (citrate synthase and OGDH) that were related to CS and AS were identified in the adrenal gland using numerous bioinformatic methods. The hub proteins were mainly enriched in mitochondrial proton transport ATP synthase complex, ATPase activation, and the AMPK signaling pathway. Compared with the control group, the adrenal glands were larger and more disordered, irregular, and necrotic in the AS+CS group. The expression of citrate synthase and OGDH was higher in the AS+CS group than in the control group, both at the protein and mRNA levels (P < 0.05). There were strong correlations among the cross-sectional areas of adrenal glands, citrate synthase, and OGDH (P < 0.05) via Spearman's rho analysis, receiver operating characteristic curves, a neural network model, and cubic spline interpolation. Enzymatic activity of citrate synthase and OGDH increased under the situation of atherosclerosis and chronic stress. Through the CCK8 assay, the adrenal cell viability was downregulated significantly after the knockdown experiment of citrate synthase and OGDH. Target genes-TF-miRNA regulatory network presented the close interrelations among the predicted microRNA, citrate synthase and OGDH. After Coimmunoprecipitation (IP) assay, the result manifested that the citrate synthase and OGDH were coexpressed in the adrenal gland. The molecular docking study showed that the docking score of optimal complex conformation between citrate synthase and OGDH was -6.15 kcal/mol. Conclusion AS combined with CS plays a significant role on the hypothalamic–pituitary–adrenal (HPA) axis, promotes adrenomegaly, increases the release of glucocorticoid (GC), and might enhance ATP synthesis and energy metabolism in the body through citrate synthase and OGDH gene targets, providing a potential research direction for future related explorations into this mechanism.
Highlights
Atherosclerosis (AS) is a chronic progressive disease characterized by the accumulation and deposition of lipid and fibrous components and an inflammatory response that leads to the development of lesions; AS is the most common and important member of a group of vascular diseases collectively called arteriosclerosis [1]
The pathological changes that occur in the adrenal gland and the possible molecular mechanisms involved in this are currently unclear in the case of AS combined with Chronic stress (CS)
The main results of this study were that citrate synthase and oxoglutarate dehydrogenase (OGDH) genes were highly expressed in adrenal tissue during AS combined with CS, and the degree of adrenal enlargement was more severe when both genes were highly expressed
Summary
Atherosclerosis (AS) is a chronic progressive disease characterized by the accumulation and deposition of lipid and fibrous components and an inflammatory response that leads to the development of lesions; AS is the most common and important member of a group of vascular diseases collectively called arteriosclerosis [1]. CS can activate various stress pathways in the body [7], including the hypothalamic–pituitary–adrenal (HPA) axis and the sympathetic nervous system (SNS) This can have adverse effects on the cardiovascular, endocrine, and immune systems, leading to the occurrence and development of stress-related diseases and threatening human health [8, 9]. Pathological changes of the adrenal gland and the possible underlying molecular mechanisms are currently unclear in the case of atherosclerosis (AS) combined with chronic stress (CS). Two most significant hub proteins (citrate synthase and OGDH) that were related to CS and AS were identified in the adrenal gland using numerous bioinformatic methods. There were strong correlations among the cross-sectional areas of adrenal glands, citrate synthase, and OGDH (P < 0:05) via Spearman’s rho analysis, receiver operating characteristic curves, a neural network model, and cubic spline interpolation.
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