首都医科大学学报 ›› 2024, Vol. 45 ›› Issue (5): 823-830.doi: 10.3969/j.issn.1006-7795.2024.05.012

• 慢性肾脏病及其并发症的发病机制及诊治 • 上一篇    下一篇

反复静脉钙刺激对慢性肾衰竭大鼠血管钙化的影响

关毅鸣,刁宗礼,黄红东,刘文虎*   

  1. 首都医科大学附属北京友谊医院肾内科, 北京 100050
  • 收稿日期:2024-08-05 出版日期:2024-10-21 发布日期:2024-10-18
  • 通讯作者: 刘文虎 E-mail:wenhuliu@mail.ccmu.edu.cn
  • 基金资助:
    国家自然科学基金项目(81570660),  北京友谊医院种子计划项目(YYZZ202114)。

Impact of repeated intravenous calcium administration on vascular calcification in chronic kidney disease rats

Guan Yiming, Diao Zongli, Huang Hongdong, Liu Wenhu*   

  1. Department of Nephrology, Beijing Friendship Hospital, Capital Medical University, Beijing 100050, China
  • Received:2024-08-05 Online:2024-10-21 Published:2024-10-18
  • Supported by:
    This study was supported by National Natural Science Foundation of China (81570660),   Beijing Friendship Hospital Seed Program  (YYZZ202114). 

摘要: 目的  探讨反复静脉钙刺激对腺嘌呤饮食诱导的慢性肾脏病(chronic kidney disease, CKD)大鼠主动脉钙化的影响。方法  选择SD大鼠32只,采用数字表法随机分为对照组和模型组,基线时检测血清肌酐、尿素氮、钙、磷水平,分别给予正常饮食或0.75%腺嘌呤饮食喂养,6周后全部更换为普通饲料。对照组和模型组大鼠再分别随机分为两组,共四组:对照+氯化钙组(Cont+CaCl2)、对照+0.9%(质量分数)氯化钠注射液(以下简称生理盐水)组(Cont+NaCl)、模型+氯化钙组(CKD+CaCl2)、模型+生理盐水组(CKD+NaCl),每组8只。分别采用隔日尾静脉注射氯化钙溶液(100 mg/kg)或等量生理盐水共8周,随后将大鼠处死,取血清检测肌酐、尿素氮、钙、磷水平;留取主动脉测定血管钙含量并进行Von Kossa染色观察有无钙化发生,采用免疫组织化学染色法观察成骨转分化指标成骨细胞特异性和转录因子(runt-related transcription factor 2, Runx2)的分布和表达情况,采用Western blotting法检测成骨转分化指标骨形态蛋白2(bone morphogenetic protein 2, BMP-2)的蛋白表达。结果  基线时各组大鼠体质量及生化指标差异无统计学意义(P>0.05);第14周结束时与对照组相比,模型组大鼠血清肌酐、尿素氮水平升高,血钙降低、血磷升高,差异有统计学意义(P<0.05);与CKD+NaCl组比较,CKD+CaCl2组血清钙、磷水平及主动脉钙含量差异无统计学意义(P>0.05),两组大鼠主动脉Von Kossa染色均为阴性;与对照组相比,模型组主动脉Runx2和BMP-2表达增加;与CKD+NaCl组比较,CKD+CaCl2组主动脉Runx2和BMP-2表达差异无统计学意义。结论  在本实验条件下,反复静脉钙刺激对腺嘌呤饮食诱导的CKD大鼠无诱发主动脉钙化作用,对主动脉成骨转分化情况无明显影响。

关键词: 慢性肾脏病, 主动脉血管钙化, 成骨转分化, 钙磷代谢紊乱

Abstract: Objective  To investigate the  impact of repeated intravenous calcium stimulation on aortic calcification in adenine diet-induced chronic kidney disease (CKD) rats.  Methods  Thirty-two Sprague-Dawley (SD) rats were randomly divided into control and model groups. Baseline serum creatinine, blood urea nitrogen, calcium, and phosphorus levels were measured. The rats were fed either a normal diet or a 0.75% adenine diet for 6 weeks, followed by a standard diet for all. Both control and model groups were further divided into two subgroups, resulting in four groups: control + calcium chloride (Cont+CaCl2), control + saline (Cont+NaCl), CKD + calcium chloride (CKD+CaCl2), and CKD + saline (CKD+NaCl), with 8 rats in each group. Tail vein injections of calcium chloride solution (100 mg/kg) or an equal volume of saline were administered every other day for 8 weeks. After the intervention, the rats were sacrificed to measure serum creatinine, blood urea nitrogen, calcium, and phosphorus levels. Aortic calcium content was assessed, and Von Kossa staining was performed to detect calcification. Immunohistochemical staining was employed to observe the distribution and expression of osteogenic differentiation markers, including the transcription factor runt-related transcription factor 2 (Runx2). Western blotting analysis was used to detect the protein expression of bone morphogenetic protein 2 (BMP-2).   Results  At baseline, there were no statistically significant differences in body weight or biochemical indicators among the groups (P>0.05). At the end of the 14th week, compared to the control group, the model group showed significantly higher serum creatinine and blood urea nitrogen levels, lower serum calcium levels, and higher serum phosphorus levels (P<0.05). There were no statistically significant differences in serum calcium, phosphorus levels, or aortic calcium content between the CKD+CaCl2 and CKD+NaCl groups (P>0.05). Von Kossa staining revealed no calcification in the aortas of either group. Compared to the control group, the model group showed increased expression of Runx2 and BMP-2 in the aorta. There were no significant differences in the expression of Runx2 and BMP-2 between the CKD+CaCl2 and CKD+NaCl groups.  Conclusions  Under the conditions of this study, repeated intravenous calcium stimulation did not induce aortic calcification in adenine diet-induced CKD rats and had no significant impact on aortic osteogenic differentiation.

Key words: chronic kidney disease, aortic vessel calcification, osteogenic differentiation, calcium-phosphorus metabolism disorder

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