中国组织工程研究

中国组织工程研究 ›› 2010, Vol. 14 ›› Issue (24): 4533-4537.doi: 10.3969/j.issn.1673-8225.2010.24.038

• 组织构建临床实践 clinical practice in tissue construction • 上一篇    下一篇

维生素D受体多态性与肠道CYP3A4表达：维生素D受体基因突变在人群中分布有差异吗？

李宝群1，郝志敏2，万丽娟1   

  1. 1 承德医学院药理教研室，河北省承德市  067000；  2承德医学院附属医院心内科，河北省承德市 067000
  • 出版日期:2010-06-11 发布日期:2010-06-11
  • 作者简介:李宝群★，女，1976年生，河北省隆化县人，满族，2007年中南大学湘雅医学院毕业，硕士，讲师，主要从事遗传药理学研究。 libao_76@163.com

Polymorphism of vitamin D receptor and expression of CYP3A4 gene in intestinal tract: Does the distribution of vitamin D receptor mutation have difference in population?

Li Bao-qun1, Hao Zhi-min2, Wan Li-juan1   

  1. 1 Department of Pharmacology, Chengde Medical University, Chengde  067000, Hebei Province, China; 2 Department of Cardiology, Affiliated Hospital of Chengde Medical University, Chengde  067000, Hebei Province, China
  • Online:2010-06-11 Published:2010-06-11
  • About author:Li Bao-qun★, Master, Lecturer, Department of Pharmacology, Chengde Medical University, Chengde 067000, Hebei Province, China libao_76@163.com
  • Supported by:

    the Subject of Education Department of Hebei Province, No. Z2009406*

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摘要：

背景：研究表明，维生素D受体可以诱导肠道中CYP3A4的转录表达，维生素D受体基因的突变将涉及到它自身所翻译的蛋白改变，而其相应的下游靶基因的转录表达或功能亦可能会发生改变。
目的：确定HT-29肠细胞中维生素D受体FokI突变在对其下游靶基因CYP3A4转录的影响。
方法：取肝胆管结石患者术中切除的肝脏组织，构建重组真核表达载体pcDNA3.1(-)B-myc/his h维生素D受体(野生型和Fok1突变型)，利用瞬时转染技术和双荧光素酶报告基因分析系统在体外HT-29细胞中检测转染不同载体后的细胞在给予不同浓度(1，10，100 nmol/L)的药物1，25(OH)2 VD3(维生素D受体的天然配体)处理后，CYP3A4的转录表达情况。
结果与结论：将野生型和突变型维生素D受体质粒共转染于 HT-29细胞中，在1，25(OH)2 VD3孵育24 h后，3个浓度代表CYP3A4mRNA转录水平的荧光素酶活性数值分别与溶媒对照组和空载体对照组相比差异具有显著性意义(P < 0.05)，且转染Fok1突变型的细胞荧光素酶活性数值稍大于转染野生型的细胞荧光素酶活性数值，但两者间差异无显著性意义(P > 0.05)。维生素D受体Fok Ⅰ突变体对其下游靶基因的转录表达与维生素D受体野生型没有差异。提示在肠细胞中，维生素D通过维生素D受体诱导CYP3A4转录表达，但是维生素D受体FokI突变体对CYP3A4的转录与野生型相比差异无显著性意义。

关键词: 维生素D受体, CYP3A4, 转录调控, 荧光素酶报告基因, 肠道

Abstract:

BACKGROUND: Studies have demonstrated that vitamin D receptor (VDR) induces CYP3A4 gene transcription and expression in tract, and the abnormity of VDR will change the quantity and quality of its protein and cause alteration of target genes transcription and expression or function in the downstream.
OBJECTIVE: To identify the effects of VDR Fok Ⅰ mutation on the transcription of CYP3A4 in intestinal tract HT-29 cells.
METHODS: Hepatic tissues were obtained from hepatolithus patients and constructed pcDNA3.1(-)B-myc/his h VDR eukaryotic expression vectors (wild type and Fok Ⅰ mutant). The HT-29 cells were transfected by using cell transfection technique and dual-luciferase report gene analytical system and treated by different concentrations of 1,25(OH)2VD3 (1, 10, and 100 nmol/L). The transcription of CYP3A4 in cultured HT-29 cells was observed.
RESULTS AND CONCLUSION: In transiently transfected HT-29 cells, the CYP3A4 luciferase activity of three concentrations (1, 10, 100 nmol/L) was increased respectively after 1,25(OH)2VD3 was added to cells for 24 hours, and there were significant differences between 1,25(OH)2VD3 group and the vehicle control group (P < 0.05), the luciferase activity from mutant VDR constructs was a little greater than wild VDR constructs, but there was no significant difference between the two VDR forms (P > 0.05). VDR Fok Ⅰ mutant could not significantly change the regulational capacity of wild type VDR on CYP3A4. The results revealed that vitamin D induces CYP3A4 expression via VDR. However, compared with wild type, the VDR Fok I mutant has no obvious difference on CYP3A4 expression.

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