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 ›› 2017, Vol. 40 ›› Issue (4): 290-296.doi: 10.3969/j.issn.1006-2157.2017.04.006

• 博士之光 • 上一篇    下一篇

六味地黄汤对慢性肾衰模型大鼠PHD2和HIF-1α表达的影响*

唐群1, 吴华2, 张熙1, 雷久士1, 蓝文蕖3, 刘春燕1#   

  1. 1 湖南中医药大学病理教研室 湖南 410208;
    2 湖南省第二人民医院芙蓉司法鉴定中心;
    3 湖南中医药大学诊断学教研室
  • 收稿日期:2016-10-17 出版日期:2017-04-30 发布日期:2017-04-30
  • 通讯作者: 刘春燕,女,博士,讲师,研究方向:慢性肾脏病的防治,E-mail:liuchunyan0221@126.com
  • 作者简介:唐群,男,博士,副教授
  • 基金资助:
    *国家自然科学基金青年项目(No.81503442、No.81603470),湖南省教育厅科研项目(No.11C0953、No.16B196)

Effects of Liuwei Dihua Tang on the expression of PHD2 and HIF-1α in rats with chronic renal failure*

TANG Qun1, WU Hua2, ZHANG Xi1, LEI Jiushi1, LAN Wenqu3, LIU Chunyan1#   

  1. 1 Department of Pathology, Hunan University of Chinese Medicine, Hunan 410208;
    2 Furong Forensic Center, Hunan Second People’s Hospital;
    3 Faculty of Diagnostics, Hunan University of Chinese Medicine
  • Received:2016-10-17 Online:2017-04-30 Published:2017-04-30

摘要: 目的 观察六味地黄汤对慢性肾衰模型大鼠肾组织中脯氨酸羟化酶2(PHD2)、缺氧诱导因子-1α(HIF-1α)表达的影响,探讨慢性肾衰的发病机制以及六味地黄汤防治慢性肾脏病的分子机制。方法 60只SD大鼠随机分为假手术组,模型组,六味地黄汤低剂量、中剂量和高剂量组。5/6肾切除法复制慢性肾衰模型。六味地黄汤低剂量、中剂量和高剂量组分别给予3.38 g/(kg·d)、6.75 g/(kg·d)、13.5 g/(kg·d)六味地黄汤灌胃。假手术组和模型组给予10 mL/(kg·d)生理盐水灌胃,每天1次,连续灌胃12周。造模12周后,检测各组大鼠24 h尿蛋白、血尿素氮和血肌酐,HE染色观察各组大鼠肾组织形态学变化,免疫组化法和RT-PCR法分别检测各组大鼠肾组织中PHD2、HIF-1α蛋白水平和基因表达。结果 模型组大鼠24 h尿蛋白、血尿素氮和血肌酐较假手术组明显升高(P<0.05),六味地黄汤低剂量、中剂量和高剂量组较模型组明显降低(P<0.05)。HE染色假手术组大鼠肾组织肾间质纤维化不明显,模型组肾间质纤维化明显,六味地黄汤低剂量、中剂量和高剂量组较模型组肾间质纤维化程度明显减轻。与假手术组比较,模型组大鼠肾组织PHD2蛋白水平和mRNA表达均明显降低(P<0.05),HIF-1α蛋白水平和mRNA表达均明显升高(P<0.05)。与模型组比较,六味地黄汤低剂量、中剂量和高剂量组大鼠肾组织中PHD2蛋白水平和mRNA表达均明显升高(P<0.05),HIF-1α蛋白水平和mRNA表达均明显降低(P<0.05),且PHD2与HIF-1α mRNA的表达呈负相关(r=-0.653,P=0.00)。结论 PHD2/HIF-1α信号途径参与肾间质纤维化,六味地黄汤可有效防治慢性肾脏病,其作用机制可能与上调PHD2、降低HIF-1α表达有关。

关键词: 六味地黄汤, 慢性肾脏病, 肾间质纤维化, 脯氨酸羟化酶2, 缺氧诱导因子-1α, 大鼠

Abstract: Objective To investigate the effects of Liuwei Dihuang Tang (LWDHT, Six-Ingredient Rehmannia Decoction) on the expression of proline Hydroxylase2 (PHD2) and hypoxia inducible factor alpha-1 (HIF-1α) in renal tissue of rats with chronic renal failure, and to investigate the pathogenesis and molecular mechanism of LWDHT in preventing and treating chronic kidney disease. Methods 60 SD rats were randomly divided into sham-operation group, model group, LWDHT low-dose, medium-dose and high-dose groups, and chronic renal failure model group (5/6 nephrectomy). The LWDHT low-, medium and high-dose groups were treated with 3.38 g/(kg·D), 6.75 g/(kg·D) and 13.5 g/(kg·D) Liuwei Dihua Decoction respectively. Sham-operation group and model group were given 10 mL/(kg·D) saline once per day for 12 weeks. At 12 weeks, 24 h urinary protein, blood urea nitrogen, and serum creatinine were measured and morphological changes of renal tissues in rats of all groups were observed with H&E staining method; protein level and gene expression of PHD2, HIF-1α in rat renal tissue were evaluated by using immunohistochemical method and RT-PCR. Results 24 h urine protein, blood urea nitrogen and serum creatinine of rats in the model group were significantly higher than those in the sham-operation group (P<0.05). These values in LWDHT low-, medium-, and high-dose groups were significantly lower than those of model group (P<0.05). HE staining results showed that renal interstitial fibrosis was not detected in sham-operation group but significant in model group; renal interstitial fibrosis was significantly reduced in the LWDHT low-, medium-, and high-dose groups. Compared with sham-operation group, levels of PHD2 protein and mRNA expression in renal tissue of model group were significantly lower (P<0.05); levels of HIF-1α protein and mRNA expression were markedly increased (P<0.05). Compared with the model group, levels of PHD2 protein and mRNA expression in rats’ kidney tissue in LWDHT low-, medium-, and high-dose groups were significantly increased (P<0.05), and expressions of HIF-1α protein and mRNA were significantly decreased (P<0.05). There was a negative correlation between the expression of PHD2 and HIF-1α mRNA (r=-0.653, P=0.00). Conclusion PHD2/HIF-1α signaling pathway seems to be involved in renal interstitial fibrosis, and Liuwei Dihuang Decoction could effectively prevent and act against chronic kidney disease. Its mechanism may be related to the up-regulation of PHD2, and down-regulation of HIF-1α.

Key words: chronic kidney disease, renal interstitial fibrosis, proline hydroxylase 2, hypoxia inducible factor 1 alpha, Liuwei Dihuang Decoction

中图分类号: 

  • R285.5