Xin yixue (Dec 2022)
Effect of DBP inhibition on vascular calcification in chronic uremic rats
Abstract
Objective To evaluate the effect of D-site binding protein （DBP） inhibition on vascular calcification in chronic uremic rats. Methods GSE146638 was downloaded from the public database （Gene Expression Omnibus， GEO）， and a total of 10 samples from rat of mRNA expression profiles were included in this study. DESeq2 was used to analyze the differentially expressed genes in the aortic smooth muscle between the control and chronic uremic rat model groups. RT-qPCR was employed to detect the expression levels of SLC22A2， ATF3， DBP and SMPD3. Male SD rats were randomly divided into four groups. In the blank group， no treatment was given. In the model group， chronic uremic rat models were established by nephrectomy of 2/3 of bilateral kidneys， and the surviving rats were fed with 8-week high phosphorus diet to induce vascular calcification. In the negative control lentivirus group （shNC group）， the rats were injected with negative control lentivirus for 4 weeks after establishing the vascular calcification uremic rat models. In the small interfering DBP group （shDBP group）， the rats were treated with siDBP lentivirus for 4 weeks after the establishment of vascular calcification uremic rat models. After corresponding interventions， the rat weight was measured， and serum creatinine， serum phosphorus， serum calcium and urea nitrogen levels were detected among four groups. Immunohistochemical staining and HE staining of the thoracic aorta tissues were used to evaluate the degree of injury. RT-qPCR and western blot were adopted to quantitatively measure the relative expression levels of Runt-related transcription factor 2 （RUNX2） and DBP. Results Compared with the control group， the rat weight was significantly less， whereas the serum creatinine levels were significantly higher in the model， shNC and shDBP groups （all P < 0.05）. In the model group， the thoracic aortic vascular wall was thickened， and the expression levels of RUNX2 and proliferating cell nuclear antigen （PCNA） were up-regulated. Compared with the model group， the serum creatinine level was declined， the expression levels of RUNX2 mRNA and protein in the thoracic aorta were down-regulated （all P < 0.05）， the expression level of PCNA in the thoracic aorta was down-regulated and the thoracic aortic vascular wall thickening was mitigated. Conclusion Inhibition of DBP may suppress vascular calcification and alleviate uremia-associated symptoms in chronic uremic rat models by down-regulating the expression levels of RUNX2 mRNA and protein in the aorta.
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