PhD Scientific Days 2023

Translational Medicine - Posters O

The Effect of Macrophages on Renal Tubular Epithelial Cells in Vitro

Előadó neve

Haghighi, Samaneh, PhD

Neptun code

KV7P20

Előadó munkahelye

Institute of Translational Medicine

Előadó telefonszáma

+36704207849

Előadó e-mail címe

haghighi_samane@yahoo.com

Az előadás címe

The Effect of Macrophages on Renal Tubular Epithelial Cells in Vitro

Szerző(k) neve és munkahelye

1 Samaneh Haghighi, Institute of Translational Medicine, Budapest
2 Anna Manzéger, Institute of Translational Medicine, Budapest
3 Gábor Kökény, Institute of Translational Medicine/International Nephrology Research and Training Center Semmelweis University, Budapest

Bemutatás módja

Poszter

Szekció

Translational Medicine - Posters O

Language of the presentation

English

Preferred session

Theoretical and Translational Medicine

Összefoglaló szövege

Introduction: Chronic kidney disease (CKD) has become an important health problem worldwide. Kidney fibrosis is the final common pathway of CKD, characterized by excessive deposition of extracellular matrix components that impair renal function, and eventually cause end-stage renal disease. Renal proximal tubular epithelial cells are very susceptible to injury. Studies have also shown the involvement of macrophages in renal fibrosis, yet the complex mechanism is poorly understood.
Aims: Thus, we aimed to investigate fibrosis-related genes and transcription factors influenced by direct activation of proximal tubular cells in vitro as compared to indirect activation by conditioned media of TGF-β or interferon-γ treated macrophages.
Methods: Human proximal tubule cell line (HK2) was cultured in DMEM supplemented with 5% FBS. Mouse macrophages (RAW 264.7. cells) were cultured in DMEM with 10% FBS. HK2 and RAW 264.7. cells were plated on 6-well plates (105 cells/well). Both cell types were treated for 24h with 10 ng/ml TGFβ and 1 U/ml IFNγ (HK2-TGF, HK2-IFN and RAW-TGF, RAW-IFN, respectively). Controls received 10 uL PBS (HK2-CTL and RAW-CTL). In order to study the effect of activated macrophages on epithelial cells, the medium of a second HK2 cell plate was replaced with conditioned medium of treated macrophages (HK2-CM-TGF and HK2-CM-IFN, respectively) and incubated for 24h (n=3/group). After the incubation, total RNA was isolated and gene expressions were assessed by qPCR. Statistical analysis was performed in SPSS 10 using Kruskal-Wallis test.
Results: TGFβ treatment reduced macrophage TGFB1 expression by 0.5-fold as compared to controls (RAW-TGF vs RAW-CTL). In contrast, the conditioned medium of TGFβ-treated macrophages induced TGFB1 expression in HK2 cells (1.6-fold). TGFβ treated HK2 cells overexpressed several pro-fibrotic factors: ACTA2 (1.7-fold), TGFβ (1.6-fold), EGR2 (3-fold) and the inflammatory CCL2 (1.7-fold) (HK2-TGF vs HK2-CTL). IFNγ induced pro-inflammatory effects on macrophages inducing STAT3 and C3 expression by 2.1-fold and 3.1-fold, respectively (RAW-IFN vs RAW-CTL). HK2 cells responded to IFNγ stimulation only with increased CCL2 expression (2.5-fold, HK2-IFN vs HK2-CTL).
Conclusion: Apart from the direct stimulatory effects of TGF-β and interferon-γ on HK2 cells, induced macrophages could also exert indirect, paracrine effects on HK2 cells via secreted substances. The exact molecular interplay between these cell types and its potential role in renal fibrosis remains to be elucidated.

University and Doctoral School

Semmelweis University, Doctoral School of Theoretical and Translational Medicine

Supervisor

Gábor Kökény

Publication of my abstract

I do not give consent to the publication of my abstract on the website of the congress.

Kind

Szabad

Status

elfogadva

Accepted presentation method

poszter

Előadás fájl jóváhagyás

nem rendelkezett róla

Előadó

7371

Start

11:24

End

11:29

Authors (legacy)

1 Samaneh Haghighi, Institute of Translational Medicine, Budapest
2 Anna Manzéger, Institute of Translational Medicine, Budapest
3 Gábor Kökény, Institute of Translational Medicine/International Nephrology Research and Training Center Semmelweis University, Budapest