主 办:北 京 中 医 药 大 学
ISSN 1006-2157 CN 11-3574/R

北京中医药大学学报 ›› 2019, Vol. 42 ›› Issue (10): 841-846.doi: 10.3969/j.issn.1006-2157.2019.10.008

• 中药药理 • 上一篇    下一篇

肾疏宁对HBV诱导人肾小管上皮细胞转分化的影响*

杨熙凯1, 徐冰2, 王耀光1#   

  1. 1 天津中医药大学第一附属医院 天津 300381;
    2 天津中医药大学
  • 收稿日期:2019-05-06 出版日期:2019-10-30 发布日期:2019-11-25
  • 通讯作者: 王耀光,男,博士,教授,主任医师,博士生导师,研究方向:中西医结合治疗肾病,E-mail:wangyaoguang1012@126.com
  • 作者简介:杨熙凯,男,在读博士生
  • 基金资助:
    *国家自然科学基金面上项目(No.81573888)

Effect of Shen Shu Ning on HBV-induced transdifferentiation of renal tubular epithelial cells*

YangXikai1,XuBing2,WangYaoguang1#   

  1. 1 First Teaching Hospital of Tianjin University of Traditional Chinese Medicine, Tianjin 300381, China;
    2 Tianjin University of Traditional Chinese Medicine, Tianjin 301617, China
  • Received:2019-05-06 Online:2019-10-30 Published:2019-11-25
  • Contact: Prof. Wang Yaoguang, Ph.D., Chief Physician, Doctoral Supervisor. First Teaching Hospital of Tianjin University of Traditional Chinese Medicine, No. 88, Changling Road, Xiqing District, Tianjin 300381, China; E-mail: wangyaoguang1012@126.com
  • Supported by:
    National Natural Science Foundation of China (No. 81573888)

摘要: 目的 探讨肾疏宁干预乙型肝炎病毒(HBV)诱导人肾小管上皮细胞(HK-2)转分化,延缓肾间质纤维化作用的可能机制。方法 通过75只C57BL/6小鼠的中药肾疏宁灌胃,制备含药血清;使用Lipofectamine转染HBV至HK-2的方法造模;将细胞分为:空白血清对照组、空质粒组、HBV组、贝那普利组、肾疏宁低浓度组、肾疏宁中浓度组、肾疏宁高浓度组。运用酶联免疫吸附实验(ELISA)、实时荧光定量PCR法、蛋白质印迹(Western blot) 法检测HK-2细胞中HBsAg、HBeAg、TGF-β1、p-Smad2、p-Smad3、Smad7、Col-Ⅲ、ColⅣ、FN的表达。结果 ELISA 检测结果:HBV组与空质粒组比较,HBsAg、HBeAg、TGF-β1表达上调(P<0.01);与HBV组比较,贝那普利组、肾疏宁低浓度组、肾疏宁中浓度组、肾疏宁高浓度组TGF-β1蛋白表达下降(P<0.001);与贝那普利组比较,肾疏宁低浓度组TGF-β1的表达相对较高(P<0.05)。实时荧光定量PCR检测结果:与HBV组比较,贝那普利组、肾疏宁各干预组ColⅢ、Col-Ⅳ、FN mRNA表达明显下降(P<0.001)。Western blot检测结果:与HBV组比较,贝那普利组、肾疏宁各干预组p-Smad2、p-Smad3蛋白表达明显下降(P<0.001)。结论 肾疏宁可通过干预TGF-β1/Smad信号通路减轻HBV诱导的人肾小管上皮细胞转分化,并减轻细胞外基质的沉积,从而延缓纤维化的进程。

关键词: 乙型肝炎病毒, 肾疏宁, 肾小管上皮细胞, TGF-β1/Smad通路

Abstract: ObjectiveTo explore the possible mechanism of Shen Shu Ning (Kidney-tonifying and Liver-soothing Formula, SSN)in the intervention of HBV-induced HK-2 transdifferentiation and delay of renal interstitial fibrosis. Methods The drug-containing serum was prepared by intragastric administration of 75 C57BL/6 mice and models were made by using lipofectamineto transfect HBV to HK-2. The cells were divided into: blank serum control group, empty plasmid group, HBV group, benazepril group, SSN low concentration group, SSN middle concentration group, and SSN high concentration group. The expressions of HBsAg, HBeAg, TGF-β1, pSmad2, pSmad3, Smad7, Col-III, Col IV and FN in HK-2 cells were detected by enzyme-linked immunosorbent assay (ELISA), real-time fluorescent quantitative PCR and Western blot. Results ELISA detection indicated that expressions of HBsAg, HBeAg and TGF-β1 were significantly increased in HBV group compared with the empty plasmid group (P<0.01). Compared with the HBV group, the expression of TGF-β1 protein was significantly decreased in the benazepril group, SSN low concentration group,SSN middle concentration group, and SSN high concentration group(P<0.001).Compared with the benazepril group, the expression of TGF-β1 in the SSN low concentration group was relatively high (P< 0.05); qPCR detection showed that, compared with the HBV group, in the benapril group, and all SSN intervention groups the expressions of Col III, Col-IV and FN mRNA were significantly decreased (P<0.001). Western blot: Compared with the HBV group, the expressions of pSmad2 and pSmad3in the benazepril group and the SSN groups were significantly decreased (P<0.001). Conclusion SSN could possibly alleviate HBV-induced transdifferentiation of human renal tubular epithelial cells and reduce the deposition of extracellular matrix by interfering with TGF-β1/Smad signaling pathway, thereby delaying the progression of fibrosis.

Key words: HBV, Shen Shu Ning(Kidney-tonifying and Liver-soothing Formula), HK-2, TGF-β1/Smad pathway

中图分类号: 

  • R285.5