天津医药

天津医药 ›› 2017, Vol. 45 ›› Issue (8): 860-864.doi: 10.11958/20170502

• 实验研究 • 上一篇    下一篇

肾康注射液对 CAPD 小鼠腹膜间皮细胞的 保护作用及机制研究

王荔 1， 夏天 1△， 张洪震 2， 李荣 1   

  1. 1 天津医科大学第二医院肾内科 （邮编 300211）， 2 检验科
  • 收稿日期:2017-04-24 修回日期:2017-06-25 出版日期:2017-08-15 发布日期:2017-08-15
  • 通讯作者: △通讯作者 E-mail：xiatian01@medmail.com.cn E-mail:19007522@qq.com
  • 作者简介:王荔 （1977）， 女， 博士， 主要从事肾脏病及腹膜透析研究
  • 基金资助:
    天津市卫计委中医中西医结合科研课题 （2015133）

Protective effect and its mechanism of Shenkang injection on peritoneal mesothelial cells in CAPD mice

WANG Li1， XIA Tian1△， ZHANG Hong-zhen2， LI Rong1   

  1. 1 Department of Nephrology, 2 Department of Clinical Laboratory, the Second Hospital of Tianjin Medical University, Tianjin 300211, China
  • Received:2017-04-24 Revised:2017-06-25 Published:2017-08-15 Online:2017-08-15
  • Contact: △Corresponding Author E-mail: xiatian01@medmail.com.cn E-mail:19007522@qq.com

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摘要： 目的 探讨肾康注射液对连续性不卧床腹膜透析 （CAPD） 小鼠腹膜间皮细胞 （PMCs） 的保护作用及可能机 制。方法 40 只 ICR 小鼠均分为空白对照（A）组， 阳性对照腹透（B）组（2.5%腹透液）， 低（C）、 中（D）、 高（E）剂量肾 康干预组（2.5%腹透液+5、 10 及 20 mL/kg 肾康注射液）， 观察至 4 周。采用生化法检测空腹血糖、 总胆固醇、 三酰甘 油、 C-反应蛋白 （CRP） 水平； 采用酶联免疫吸附试验检测血清及透出液中肿瘤坏死因子-α （TNF-α）、 转化生长因子- β1 （TGF-β1）、 血管内皮生长因子 （VEGF）、 结缔组织生长因子 （CTGF） 水平； HE 染色观察腹膜组织病理改变； 分别以 免疫组化染色和 Real-time PCR 检测腹膜组织中 TNF-α、 TGF-β1、 VEGF、 CTGF 的蛋白质表达和 mRNA 转录水平。 结果 各组小鼠体质量、 空腹血糖、 总胆固醇、 三酰甘油水平差异无统计学意义。与 B 组比较， C、 E 组差异均无统计 学意义， 但 D 组 CRP 水平显著减低， 无论是血清还是腹腔透出液中， C、 D 组 TNF-α、 TGF-β1、 VEGF 及 CTGF 表达水 平均降低， 而 E 组 TNF-α、 TGF-β1 升高 （P<0.05）， 但 E 组 VEGF 及 CTGF 差异均无统计学意义。与 E 组比较， 除 C 组腹腔液中 CTGF 差异无统计学意义外， 血清及腹腔透出液中 C、 D 组 TNF-α、 TGF-β1、 VEGF、 CTGF 均降低（P< 0.05）。B 组可见腹膜间皮细胞均有损害； C、 D、 E 组有不同程度改善。与 B 组比较， C、 D、 E 组 TNF-α、 TGF-β1、 VEGF、 CTGF 蛋白及 mRNA 水平呈逐渐降低趋势 （P＜0.05）。结论 一定浓度的肾康注射液可通过抑制 CAPD 小鼠 PMCs 的 TNF-α、 TGF-β1、 VEGF、 CTGF 的表达， 保护 PMCs， 从而控制腹膜纤维化的发生发展。

关键词: 腹膜透析, 持续不卧床, 肾功能衰竭, 腹膜后纤维化, 疾病模型, 动物, 腹膜间皮细胞, 肾康注射液

Abstract: Objective To study the protective effect of Shenkang injection on peritoneal mesothelial cells (PMCs) of continuous ambulatory peritoneal dialysis (CAPD) mice, and explore the possible mechanism. Methods Forty ICR mice were randomly divided into blank control (A) group, peritoneal dialysis (B) group, low dose of Shenkang (C) group (2.5% dialysate + 5 mL/kg Shenkang injection), medium (D) group (2.5% dialysate + 10 mL/kg Shenkang injection) and high (E) group (2.5% dialysate + 20 mL/kg Shenkang injection). Mice were observed for 4 weeks. Fasting blood glucose, total cholesterol, triglyceride and C- reactive protein (CRP) were detected by biochemical assay. Tumor necrosis factor- alpha (TNF- α), transforming growth factor- beta 1 (TGF- β1), vascular endothelial growth factor (VEGF) and connective tissue growth factor (CTGF) levels of serum and dialysate were detected by ELISA. Pathological changes of peritoneal tissue were observed by HE staining. Expression and mRNA transcription levels of these four cytokines in the peritoneal tissue were detected by immunohistochemical staining and real-time PCR respectively. Results There were no significant differences in body weight, fasting blood glucose, total cholesterol and triglyceride between 5 groups of mice (P>0.05). Compared with group B, there was no significant difference in CRP level between group C and group E, but which was significantly decreased in group D (P<0.05). The serum and dialysate levels of TNF-α, TGF-β1, VEGF and CTGF were decreased in group C and group D. The serum and dialysate levels of TNF-α and TGF-β1 were significantly increased in group E (P< 0.05), but there was no significant difference between VEGF and CTGF in group E. Compared with group E, except for CTGF in dialysate of group C, the serum and dialysate levels of TNF-α, TGF-β1, VEGF and CTGF were significant decreased in group C and group D (P<0.05). Damaged PMCs were found in group B, which were improved in various degrees in group C, group D and group E. Compared with group B, the protein expression and mRNA relative transcription levels of TNF- α, TGF-β1, VEGF and CTGF tended to decrease gradually in group C, group D and group E (P<0.05). Conclusion A certain concentration of Shenkang injection can protect PMCs by inhibiting the expression of TNF-α, TGF-β1, VEGF and CTGF in CAPD mice, so as to control the occurrence and development of peritoneal fibrosis.

Key words: peritoneal dialysis, continuous ambulatory, kidney failure, retroperitoneal fibrosis, disease models, animal, peritoneal mesothelial cells, Shenkang injection