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Human NPC1L1 Expression is Positively Regulated by PPARα

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ABSTRACT

Purpose

Niemann-Pick C1-like 1 (NPC1L1), a pharmacological target of ezetimibe, is responsible for cholesterol absorption in enterocytes and hepatocytes. In the present study, the involvement of peroxisome proliferator-activated receptor α (PPARα) and its cofactor, PPARγ coactivator 1α (PGC1α) in the transcriptional regulation of human NPC1L1 was analyzed.

Methods

Reporter gene assays and electrophoretic mobility shift assays (EMSAs) were performed with the 5′-flanking region of the human NPC1L1 gene and the effect of siPPARα was examined.

Results

PPARα-mediated transactivation was observed with human NPC1L1 promoter constructs. Detailed analyses using deletion- and mutated-promoter constructs revealed the presence of a functional PPARα-response element (PPRE) upstream of the human NPC1L1 gene (−846/−834), a direct binding of PPARα and RXRα to which was confirmed by EMSAs. Moreover, PPARα-specific knockdown resulted in a significant decrease in the endogenous expression of NPC1L1 mRNA and protein in human-derived HepG2 cells. Furthermore, cotransfection of PGC1α stimulated the SREBP2/HNF4α- and PPARα/RXRα-mediated activation of the human NPC1L1 promoter.

Conclusions

We found that PPARα positively regulates human NPC1L1 transcription via direct binding to a PPRE. Additionally, PGC1α stimulates the SREBP2/HNF4α- and PPARα/RXRα-mediated transactivation of human NPC1L1. These findings may provide new insights into the close relationship of glucose, fatty acids and cholesterol homeostasis.

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Abbreviations

BSA:

bovine serum albumin

DMEM:

Dulbecco’s Modified Eagle Medium

DR-1:

direct repeat-1

EMSA:

electrophoretic mobility shift assay

HNF4α:

hepatocyte nuclear factor 4α

I-FABP:

intestinal-fatty acid binding protein

NPC1L1:

Niemann-Pick C1-like 1

PGC1α:

PPARγ coactivator 1α

PPAR:

peroxisome proliferator-activated receptor

PPRE:

PPARα-response element

RXR:

retinoid X receptor

SDS:

sodium dodecyl sulfate

SREBP2:

sterol responsive element binding protein 2

TBS-T:

Tris-buffered saline containing 0.05% Tween 20

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ACKNOWLEDGMENTS

This work was supported by grants from the Ministry of Education, Culture, Sports, Science and Technology of Japan.

Author information

Author notes

  1. Tomohiro Terada

    Present address: Department of Hospital Pharmacy, Shiga University of Medical Science, Otsu, 520-2192, Japan

  2. Ken-ichi Inui

    Present address: Kyoto Pharmaceutical University, Yamashina-ku, Kyoto, 607-8414, Japan

Authors and Affiliations

  1. Department of Pharmacy, The University of Tokyo Hospital Faculty of Medicine, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo, 113-8655, Japan

    Yuki Iwayanagi, Tappei Takada, Fumiya Tomura, Yoshihide Yamanashi & Hiroshi Suzuki

  2. Department of Pharmacy, Kyoto University Hospital Faculty of Medicine, Kyoto University, Sakyo-ku, Kyoto, 606-8507, Japan

    Tomohiro Terada & Ken-ichi Inui

Authors
  1. Yuki Iwayanagi
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  2. Tappei Takada
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  3. Fumiya Tomura
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  4. Yoshihide Yamanashi
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Corresponding author

Correspondence to Tappei Takada.

Additional information

Yuki Iwayanagi and Tappei Takada contributed equally to this work.

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Iwayanagi, Y., Takada, T., Tomura, F. et al. Human NPC1L1 Expression is Positively Regulated by PPARα. Pharm Res 28, 405–412 (2011). https://doi.org/10.1007/s11095-010-0294-4

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  • DOI: https://doi.org/10.1007/s11095-010-0294-4

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