吡咯烷二硫代甲酸铵（PDTC）抑制尿毒症大鼠主动脉钙化的研究

发布时间：2018-05-11 22:02

本文选题：尿毒症 + 血管钙化　；参考：《天津医科大学》2014年硕士论文

【摘要】：目的：心血管疾病(cardiovascular disease, CVD)是终末期肾脏病(end-stage renal disease, ESRD)的主要死因,大约50%ESRD患者死于CVD,是普通人群的20-30倍。因此,有关ESRD心血管并发症的原因及防治措施一直是国内外学者探讨的的重点课题。血管钙化是ESRD患者发生CVD的独立危险因素。现有研究认为,影响CKD患者血管钙化的因素除了传统因素(如高血压、脂代谢异常、糖尿病,高龄、吸烟)外,还包括非传统因素,如钙磷代谢异常、氧化应激、炎症介质等。NF-κB是炎症的重要介质。已经证实,NF-κB参与了尿毒症血管钙化过程。本研究旨在探讨NF-κB抑制剂吡咯烷二硫代甲酸铵(PDTC)对尿毒症大鼠主动脉钙化的干预作用及相关机制。方法：16只雄性Sprague-Dawley大鼠,随机分为尿毒症模型组及PDTC干预组,均以0.75%腺嘌呤及高磷饮食喂养,制备尿毒症动脉钙化模型,干预组同时腹腔注射PDTC100mg/kg/d。另取8只匹配大鼠作为健康对照组。8周后处死大鼠,HE及von Kossa染色观察腹主动脉病理改变及钙化情况,免疫组织化学方法检测OPN、Cbfa-1在主动脉的定位表达,Western印迹检测主动脉NF-κB总p65与细胞核p-p65、骨桥蛋白(OPN)、核心结合因子al (Cbfa-1)蛋白表达。结果：造模4周及8周,尿毒症组、PDTC干预组大鼠血清Scr、血磷、钙磷乘积均显著高于对照组(均P0.01),但此两组间差异无统计学意义(P0.05)；给药8周,主动脉均出现明显钙化,以中膜钙化为主,但PDTC干预组程度较轻(P0.05)；免疫组化显示,大鼠主动脉OPN表达于中膜及外膜,Cbfa-1表达于中膜细胞核,二者在PDTC干预组表达均较尿毒症组下降(均P0.05)；Western印迹结果显示,PDTC干预组主动脉NF-κB总p65、核p-p65、OPN、Cbfa-1表达均较尿毒症组下降(均P0.01),且Cbfa-1表达与p65、核p-p65表达均呈正相关(r=0.707, P=0.000; r=0.507, P=0.000)。结论：PDTC可部分阻断NF-κB p65核转位所致尿毒症大鼠血管平滑肌细胞向成骨细胞转分化及主动脉钙化。
[Abstract]:Objective: cardiovascular disease (CVD) is the main cause of death of end-stage renal disease, ESRD) in end-stage renal disease, about 20-30 times of that in the general population. Therefore, the causes and preventive measures of cardiovascular complications in ESRD have been the focus of domestic and foreign scholars. Vascular calcification is an independent risk factor for CVD in patients with ESRD. Current studies suggest that the factors affecting vascular calcification in patients with CKD include non-traditional factors, such as abnormal calcium and phosphorus metabolism, oxidative stress, in addition to traditional factors (such as hypertension, abnormal lipid metabolism, diabetes, old age, smoking). NF- 魏 B is an important mediator of inflammation. NF- 魏 B has been confirmed to be involved in the vascular calcification process in uremia. The aim of this study was to investigate the effect of NF- 魏 B inhibitor, ammonium pyrrolidine dithiocarbamate, on aortic calcification in uremic rats. Methods Sixteen male Sprague-Dawley rats were randomly divided into two groups: uremic model group and PDTC intervention group. The rats were fed with 0.75% adenine and high phosphorus diet to establish the model of arterial calcification of uremia. The intervention group was injected with PDTC 100 mg / kg / d intraperitoneally at the same time. In addition, 8 matched rats were used as control group. After 8 weeks, the rats were killed to observe the pathological changes and calcification of abdominal aorta by HE and von Kossa staining. The expression of OPN- Cbfa-1 in aorta was detected by immunohistochemical method. Western blot was used to detect the expression of total p65 of NF- 魏 B and p-p65 in nucleus, OPN of osteopontin and Cbfa-1). Results: after 4 and 8 weeks of modeling, the serum Scrs, serum phosphorus, calcium and phosphorus products in the uremic group were significantly higher than those in the control group (P 0.01), but there was no significant difference between the two groups (P 0.05), and at 8 weeks of administration, calcification was observed in the aorta. Medial calcification was predominant, but the degree of PDTC intervention group was mild (P0.05A), and the expression of OPN in rat aorta was found in the medial membrane and adventitia Cbfa-1 in the nucleus of the mesangium, and the expression of Cbfa-1 in the rat aorta was observed by immunohistochemistry. The expression of NF- 魏 B in aorta of PDTC intervention group was lower than that of uremia group (P 0.05). The expression of Cbfa-1 in nucleus p-p65 was significantly lower than that in uremia group (P 0.01), and the expression of Cbfa-1 was positively correlated with p65. The expression of nuclear p-p65 was positively correlated with that of P0. 707, P0. 000, rn 0. 507, P0. 000. Conclusion the osteoblast transdifferentiation and aortic calcification of vascular smooth muscle cells induced by nuclear translocation of NF- 魏 B p65 in rats with uremia were partially blocked by 1: PDTC.
【学位授予单位】：天津医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R692.5

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