, Volume 68, Issue 4, pp 1337–1348 | Cite as

PPARγ ameliorated LPS induced inflammation of HEK cell line expressing both human Toll-like receptor 4 (TLR4) and MD2

  • Reyhaneh Darehgazani
  • Maryam Peymani
  • Motahare-Sadat Hashemi
  • Mir Davood Omrani
  • Abolfazl Movafagh
  • Kamran Ghaedi
  • Mohammad Hossein Nasr-Esfahani
Original Article


TLR4 is transmembrane pattern-recognition receptor that initiates signals in response to diverse pathogen-associated molecular patterns especially LPS. Recently, there have been an increasing number of studies about the role of TLRs in the pathogenesis of several disorders as well as the therapeutic potential of TLR intervention in such diseases. Peroxisome proliferator-activated receptor-gamma (PPARγ) is a ligand-activated transcription factor with numerous biological effects. PPARγ has been shown to exert a potential anti-inflammatory effect through suppression of TLR4-mediated inflammation. Therefore, PPARγ agonists may have a potential to combat inflammatory conditions in pathologic states. The current study aims to show the decrease of inflammation by overexpression of PPARγ in a cell reporter model. To reach this goal, recombinant pBudCE4.1 (+) containing encoding sequences of human TLR4 and MD2 was constructed and used to transfect HEK cells. Subsequently, inflammation was induced by LPS treatment as control group. In the treatment group, overexpression of PPARγ prior to inflammation was performed and the expression of inflammatory markers was assessed in this condition. The expression of inflammatory markers (TNFα and iNOS) was defined by quantitative real time PCR and the amount of phosphorylated NF-κB was measured by western blot. Data indicated expression of TNFα and iNOS increased in LPS induced inflammation of stably transformed HEK cells with MD2 and TLR4. In this cell reporter model overexpression of PPARγ dramatically prevented LPS-induced inflammation through the blocking of TLR4/NF-κB signaling. PPARγ was shown to negatively regulate TLR4 activity and therefore exerts its anti-inflammatory action against LPS induced inflammation.


HEK Inflammation MD2 PPARγ TLR4 



Bovine serum albumin


Coding sequence




Dulbecco’s modified Eagle’s medium


Elongation factor 1α-subunit


Fetal bovine serum


Glyceraldehyde 3-phosphate dehydrogenase


Human embryonic kidney






Leucine-rich repeat


Myeloid differentiation protein 2


3-(4,5-Dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium)/phenazine methosulfate


Oligodendrocyte precursor cell


Pathogen-associated molecular patterns


Phosphate buffered saline


Pattern-recognition receptor




Toll-like receptor


Conflict of interest

None of the authors has any conflicts of interest to disclose and all authors support submission to this journal.


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Copyright information

© Springer Science+Business Media Dordrecht 2015

Authors and Affiliations

  • Reyhaneh Darehgazani
    • 1
    • 2
  • Maryam Peymani
    • 2
    • 3
  • Motahare-Sadat Hashemi
    • 2
  • Mir Davood Omrani
    • 1
  • Abolfazl Movafagh
    • 1
  • Kamran Ghaedi
    • 2
    • 4
  • Mohammad Hossein Nasr-Esfahani
    • 2
  1. 1.Department of Medical Genetics, Faculty of MedicineShahid Beheshti University of Medical SciencesTehranIran
  2. 2.Department of Cellular Biotechnology, Cell Science Research CenterRoyan Institute for BiotechnologyIsfahanIran
  3. 3.Department of Biology, Faculty of Basic Sciences, Shahrekord BranchIslamic Azad UniversitySahrekordIran
  4. 4.Biology Department, School of SciencesUniversity of IsfahanIsfahanIran

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