抑制DBP对慢性尿毒症大鼠血管钙化的影响  

Effect of DBP inhibition on vascular calcifi cation in chronic uremic rats

作　　者：曹素娟 刘书政 袁步奇 袁培乐 赵松鹤 张云芳 Cao Sujuan;Liu Shuzheng;Yuan Buqi;Yuan Peile;Zhao Songhe;Zhang Yunfang(Department of Thyroid Surgery,Huadu District People’s Hospital,Southern Medical University,Guangzhou 510800,China;不详)

机构地区：[1]南方医科大学附属花都医院广州市花都区人民医院甲状腺外科,广州510800 [2]南方医科大学附属花都医院广州市花都区人民医院肾病科,广州510800

出　　处：《新医学》2022年第12期908-913,共6页Journal of New Medicine

基　　金：广州市科学技术局基础与应用基础研究项目(202002030285)。

摘　　要：目的探讨转录因子D位点结合蛋白(DBP)基因修饰对慢性尿毒症大鼠血管钙化的影响。方法从基因表达综合数据库(GEO)下载表达谱数据GSE146638,共纳入10只大鼠样本的mRNA表达谱,采用DESeq2差异表达分析数据库中正常对照组和慢性尿毒症组大鼠主动脉血管平滑肌的差异表达基因,记录逆转录-实时荧光定量PCR(RT-qPCR)检测的上调基因糖转运蛋白基因SLC22A2、转录激活因子3(ATF3)、DBP、鞘磷脂磷酸二酯酶3(SMPD3)的表达水平。另选择24只雄性SD大鼠分为4组,空白组不做任何处理;模型组切除2/3左肾和右肾诱导慢性尿毒症模型,存活大鼠喂饲高磷饲料8周诱导血管钙化;慢病毒对照组(shNC组)在建立慢性尿毒症大鼠血管钙化模型后注射对照慢病毒治疗4周;小干扰DBP(siDBP)慢病毒实验组(shDBP组)在建立慢性尿毒症大鼠血管钙化模型后注射siDBP慢病毒治疗4周。干预后测量4组大鼠的体质量,检测4组大鼠的血清肌酐、血钙、血磷和尿液尿素氮水平,并用胸主动脉组织HE染色和免疫组织化学染色评估模型的损伤程度;应用RT-qPCR和蛋白免疫印迹法检测大鼠主动脉中Runt相关转录因子2(RUNX2)以及DBP的相对表达量。结果与空白组比较,模型组、shNC组和shDBP组大鼠的体质量较低,血清肌酐水平较高(P均<0.05);模型组大鼠胸主动脉血管壁变厚、RUNX2和增殖细胞核抗原(PCNA)表达上调;与模型组比较,shDBP组大鼠血清肌酐水平下降、胸主动脉RUNX2 mRNA和蛋白表达均降低(P均<0.05),胸主动脉PCNA表达减弱,胸主动脉壁增厚程度减轻。结论下调主动脉血管中的RUNX2 mRNA和蛋白表达抑制DBP表达,可能减轻慢性尿毒症大鼠血管钙化程度和尿毒症相关症状。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 SLC22 A2,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 lenti virus 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

关 键 词：慢性尿毒症 血管钙化 D位点结合蛋白 Runt相关转录因子2 

分 类 号：R692.5[医药卫生—泌尿科学]