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 ›› 2018, Vol. 41 ›› Issue (1): 60-64.doi: 10.3969/j.issn.1006-2157.2018.01.010

• 科技之窗 • 上一篇    下一篇

六味地黄汤含药血清对HK-2细胞PHD2/HIF-1α信号途径的影响*

唐群1, 吴华2#, 刘春燕1, 罗琳1, 易纯1, 徐文峰3, 庄高建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 湖南中医药大学第一附属医院肾脏内科;
  • 收稿日期:2017-07-28 出版日期:2018-01-30 发布日期:2018-01-30
  • 通讯作者: 吴华,女,硕士,副主任法医师,研究方向:法医病理教学与科研,E-mail:aqiaowu@163.com
  • 作者简介:唐群,男,博士,副教授,硕士生导师
  • 基金资助:
    *国家自然科学基金项目(No. 81503442,No. 81603470),湖南省教育厅科研项目(No. 17B197,No. 16B196)

Influence of medicated serum of Liuwei Dihuang Tang on PHD2/HIF-1α signal pathway in HK-2 cells*

Tang Qun1, Wu Hua2#, Liu Chunyan1, Luo Lin1, Yi Chun1, Xu Wenfeng3, Zhuang Gaojian1   

  1. 1 Department of Pathology, Hunan University of Chinese Medicine, Hunan 410208, China;
    2 Furong Judicial Identification Center, Hunan Second People’s Hospital, Hunan 410007, China;
    3 Department of Nephrology, First Affiliated Hospital of Hunan University of Chinese Medicine, Hunan 410007, China;
  • Received:2017-07-28 Online:2018-01-30 Published:2018-01-30
  • Supported by:
    National Natural Science Foundation of China (No. 81503442、No. 81603470), Education Department of Hunan Province (No. 17B197, No. 16B196)

摘要: 目的 以缺氧诱导的HK-2细胞为研究对象,探讨脯氨酸羟化酶2/缺氧诱导因子1α(PHD2/HIF-1α)信号途径对肾间质纤维化的影响,以及六味地黄汤含药血清干预作用。方法 40只SPF级雄性SD大鼠分为空白组和给药组,每组20只,空白组以10 mL/(kg·d)生理盐水,给药组以67.5 g/(kg·d)六味地黄汤灌胃连续1周,腹主动脉取血,离心后分离血清。体外培养HK-2细胞,将实验分为空白对照组(10%胎牛血清)、正常血清组(10%正常大鼠血清)、氯化钴(CoCl2)处理组(150 μmol/LCoCl2+10%正常大鼠血清)和含药血清组(150 μmol/LCoCl2+10%六味地黄汤含药血清)。体外培养HK-2,分组处理24 h后,用Western Blot法和RT-PCR法分别检测六味地黄汤含药血清对缺氧诱导的HK-2细胞中PHD2、HIF-1α、结缔组织生长因子(CTGF)的蛋白水平和mRNA表达的影响。结果 与正常血清组比较,CoCl2处理组HK-2细胞中PHD2的蛋白水平和mRNA表达均降低(P<0.05),而HIF-1α、CTGF的蛋白水平和mRNA表达均升高(P<0.05);与CoCl2处理组比较,含药血清组HK-2细胞中PHD2的蛋白水平和mRNA表达均升高(P<0.05),而HIF-1α、CTGF的蛋白水平和mRNA表达均降低(P<0.05)。结论 PHD2/HIF-1α信号途径参与肾间质纤维化,六味地黄汤可能通过上调PHD2的表达,促进HIF-1α的降解,后者下调CTGF的表达,从而抑制肾间质纤维化。

关键词: HK-细胞, 肾间质纤维化, 脯氨酸羟化酶2, 缺氧诱导因子1α, 结缔组织生长因子, 六味地黄汤含药血清, 大鼠

Abstract: Objective To investigate the effect of PHD2/HIF-1 signaling pathway on the renal interstitial fibrosis, and the intervention effect of medicated serum of Liuwei Dihuang Tang on HK-2 cells induced by hypoxia. Methods A total of 40 SPF male SD rats were divided into blank group and medicated group (each n=20). The blank group was orally given normal saline 〔10 mL/(kg·d)〕 and medicated group was orally given Liuwei Dihuang Tang for 1 week continuously, and then blood samples were collected from rat abdominal aorta for separating serum after centrifugation. HK-2 cells were cultured in vitro. HK-2 cells were divided into blank control group (10% of fetal bovine serum), normal group (10% of normal serum), CoCl2 group (150 μmol/L of CoCl2+10% of normal serum) and medicated serum group (150 μmol/L of CoCl2+10% of medicated serum of Liuwei Dihuang Tang). HK-2 cells were cultivated in vitro and treated in all groups for 24 h, and then the influence of medicated serum of Liuwei Dihuang Tang on protein levels and mRNA expressions of PHD2, HIF-1α and connective tissue growth factor (CTGF) in HK-2 cells were detected, respectively, by using Western blotting method and reverse transcription polymerase chain reaction (RT-PCR). Results Compared with normal group, the protein level and mRNA expression of PHD2 in HK-2 cells all decreased (P<0.05), while protein levels of HIF-1α and CTGF and mRNA expressions of HIF-1α and CTGF increased (P<0.05) in CoCl2 group. Compared with CoCl2 group, the protein level and mRNA expression of PHD2 increased (P<0.05), and those of HIF-1α and CTGF decreased (P<0.05) in medicated serum group. Conclusion PHD2/HIF-1α signal pathway takes part in RIF, and Liuwei Dihuang Tang can inhibit RIF through up-regulating PHD2 expression, improving HIF-1α degradation and down-regulating CTGF expression.

Key words: HK-2 cells, renal interstitial fibrosis, proline hydroxylase 2, hypoxia-inducible factor 1α, connective tissue growth factor, medicated serum of Liuwei Dihuang Tang (Six-Ingredient Rehmannia Decoction) , rats

中图分类号: 

  • R285.5