天津医药

天津医药 ›› 2021, Vol. 49 ›› Issue (7): 699-705.doi: 10.11958/20203042

• 实验研究 • 上一篇    下一篇

扶肾方对尿毒症腹膜透析大鼠腹膜超滤功能及VEGF-Notch信号通路的影响

杨波，王孟孟，孙林，钟柯，李洁，杨洪涛   

  1. 1天津中医药大学第一附属医院肾病科（邮编300381）；2安徽省阜阳市第四人民医院内分泌科
  • 收稿日期:2020-11-05 修回日期:2021-01-13 出版日期:2021-07-15 发布日期:2021-07-12
  • 作者简介:杨波（1982），男，博士，副主任医师，主要从事中西医结合防治慢性肾脏病及中医药提高肾透析患者生存质量研究。E-mail：yb8203@126.com
  • 基金资助:
    国家自然科学基金面上项目（81774065）；国家自然科学基金青年基金（81303092）

The effect of Fushen recipe on peritoneal ultrafiltration function and VEGF-Notch signal pathway in uremic peritoneal dialysis rats

YANG Bo, WANG Meng-meng, SUN Lin, ZHONG Ke, LI Jie, YANG Hong-tao   

  1. 1 Department of Nephrology, the First Teaching Hospital of Tianjin University of Traditional Chinese Medicine, Tianjin 300381, China; 2 Department of Endocrinology, Fuyang Fourth People's Hospital, Anhui Province
  • Received:2020-11-05 Revised:2021-01-13 Published:2021-07-15 Online:2021-07-12

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摘要： 目的　探讨扶肾方对尿毒症腹膜透析大鼠的腹膜超滤功能及腹膜组织Notch1、delta-样配体4（Dll4）、Notch胞内域（NICD）、Hes1、血管内皮生长因子（VEGF）及血管内皮生长因子受体-2（VEGFR-2）的影响。方法　采用随机数字表法将50只雄性SD大鼠分为正常组、模型组、扶肾方低剂量组（模型+扶肾方324 g/L灌胃）、扶肾方高剂量组（模型+扶肾方648 g/L灌胃）和塞来昔布组（模型+塞来昔布1.8 g/L灌胃），每组10只。规律腹腔注射腹膜透析液4周，行腹膜平衡实验计算超滤量后处死大鼠，取腹膜组织，HE染色观察腹膜形态学变化，采用实时荧光定量PCR（qPCR）和蛋白印迹法检测Notch1、Dll4、NICD、Hes1、VEGF、VEGFR-2的mRNA和蛋白表达，蛋白印迹法检测p-VEGFR-2的蛋白水平。结果　模型组、扶肾方低剂量组、扶肾方高剂量组及塞来昔布组大鼠腹膜超滤量明显低于正常组，扶肾方高剂量组与塞来昔布组超滤量高于模型组和扶肾方低剂量组，塞来昔布组超滤量低于扶肾方高剂量组（P＜0.05）。腹膜HE染色显示，扶肾方低剂量组与高剂量组大鼠腹膜新生血管较模型组明显减少。与正常组比较，模型组大鼠腹膜组织Notch1、Dll4、NICD、Hes1 mRNA及蛋白表达均下调，VEGFR-2、VEGF mRNA及VEGFR-2、p-VEGFR-2、VEGF蛋白表达上调（P＜0.05）。与模型组比较，扶肾方低剂量组NICD、Hes1 mRNA及Notch1、Dll4、NICD、Hes1蛋白表达上调，VEGFR-2、VEGF mRNA及VEGFR-2、p-VEGFR-2、VEGF蛋白表达下调（P＜0.05）；扶肾方高剂量组Notch1、Dll4、NICD、Hes1 mRNA及Dll4、Hes1蛋白表达上调，VEGFR-2、VEGF mRNA及VEGFR-2、p-VEGFR-2、VEGF蛋白表达下调（P＜0.05）；塞来昔布组Notch1 mRNA及Notch1、Dll4蛋白表达上调，VEGFR-2、VEGF mRNA及p-VEGFR-2、VEGFR-2、VEGF蛋白表达下调（P＜0.05）。结论　扶肾方可改善腹膜超滤功能，该作用可能与上调腹膜组织Notch1、Dll4、NICD、Hes1 mRNA及蛋白表达，下调VEGFR-2、VEGF mRNA及VEGFR-2、p-VEGFR-2、VEGF蛋白表达有关。

关键词: 腹膜透析, 尿毒症, 超滤, 血管内皮生长因子类, 受体, Notch, 扶肾方, VEGF-Notch信号通路

Abstract: Objective　To explore the effects of Fushen recipe on the peritoneal ultrafiltration function and Notch1, delta- like ligand 4 (Dll4), Notch intracellular domain (NICD), hairy and enhancer of split homolog-1 (Hes1), vascular endothelial growth factor (VEGF) and vascular endothelial growth factor receptor-2 (VEGFR-2) in peritoneal tissues of uremic peritoneal dialysis rats. Methods　Fifty male SD rats were randomly divided into 5 groups: normal group, model group, Fushen recipe low dose group (model + Fushen recipe 324 g/L), Fushen recipe high dose group (model + Fushen recipe 648 g/L) and Celecoxib group (model + Celecoxib 1.8 g/L), with 10 rats in each group. The peritoneal dialysis solution was injected intraperitoneally regularly for 4 weeks. The rats were killed after the calculation of ultrafiltration volume by peritoneal balance test. The peritoneal tissues were taken for the observation of morphological changes of peritoneum by HE staining. The mRNA and protein expressions of Notch1, Dll4, NICD, Hes1, VEGF and VEGFR-2 were detected by real-time fluorescence quantitative PCR (qPCR) and Western blot assay, and the protein level of p-VEGFR-2 was detected by Western blot assay. Results　The peritoneal ultrafiltration volume was significantly lower in model group, Fushen recipe low-dose group, Fushen recipe high-dose group and Celecoxib group than that in normal group, while the peritoneal ultrafiltration volume was significantly higher in Fushen recipe high-dose group and Celecoxib group than that of model group and Fushen recipe low-dose group. The peritoneal ultrafiltration volume was significantly lower in Celecoxib group than that of Fushen recipe high-dose group (P＜0.05). The peritoneal HE staining showed that peritoneal neovascularization was significantly reduced in Fushen recipe low-dose and high-dose groups compared with that in model group. Compared with normal group, the expressions of Notch1, NICD, Dll4, Hes1 mRNA and protein were down-regulated in peritoneal tissue of model group, while the expressions of VEGFR-2, VEGF mRNA and VEGFR-2, p-VEGFR-2 and VEGF protein were significantly up-regulated (P＜0.05). Compared with the model group, the expressions of NCID, Hes1 mRNA and Notch1, NICD, Dll4 and Hes1 protein were up-regulated, while the expressions of VEGFR-2, VEGF mRNA and VEGFR-2, p-VEGFR-2 and VEGF protein were down-regulated in the Fushen recipe low-dose group (P＜0.05). The expressions of Notch1, NICD, Dll4, Hes1 mRNA and Dll4, Hes1 protein were up-regulated, while the expressions of VEGFR-2, VEGF mRNA and VEGFR-2, p-VEGFR-2, VEGF protein were significantly down-regulated (P＜0.05). In Celecoxib group, Notch1 mRNA and Notch1, Dll4 protein were up-regulated, while VEGFR-2 and VEGF mRNA and p-VEGFR2, VEGFR-2 and VEGF protein were significantly down-regulated (P＜0.05). Conclusion　Fushen recipe can improve peritoneal ultrafiltration function, which may be related to the up-regulation of Notch1, NICD, Dll4, Hes1 mRNA and protein expression in peritoneal tissue, and the down-regulation of VEGFR-2, VEGF mRNA and VEGFR-2, p-VEGFR-2 and VEGF protein expression.

Key words: peritoneal dialysis, uremia, ultrafiltration, vascular endothelial growth factors, receptors, Notch, Fushen recipe, VEGF-Notch signaling pathway