TUNICAMYCIN INDUCED SPHINGOLIPID AND POLYUNSATURATED FATTY ACID CHANGES IN HUMAN KIDNEY EPITHELIAL CELLS

Aydın Aslan M.

TURKISH JOURNAL OF BIOCHEMISTRY-TURK BIYOKIMYA DERGISI, cilt.46, ss.100-104, 2021 (SCI-Expanded)

  • Yayın Türü: Makale / Özet
  • Cilt numarası: 46
  • Basım Tarihi: 2021
  • Dergi Adı: TURKISH JOURNAL OF BIOCHEMISTRY-TURK BIYOKIMYA DERGISI
  • Derginin Tarandığı İndeksler: Science Citation Index Expanded (SCI-EXPANDED), Scopus, Academic Search Premier, EMBASE, Food Science & Technology Abstracts, TR DİZİN (ULAKBİM)
  • Sayfa Sayıları: ss.100-104
  • Akdeniz Üniversitesi Adresli: Evet

Özet

BACKGROUND AND AIM: This study aimed to determine tunicamycin

(TM)-related sphingolipid and polyunsaturated fatty acid (PUFA) changes

in human kidney cells (HEK-293) and their effects on inflammatory and

apoptotic pathways. Tunicamycin was employed to induce endoplasmic

reticulum (ER) stress in HEK-293 cells and an ER stress inhibitor,

tauroursodeoxycholic acid (TUDCA), was given to minimize cytotoxicity.

METHODS: Cell viability was determined by MTT assay. Sphingomyelin (SM),

ceramide (CER) and PUFA levels were measured by liquid chromatography-tandem

mass spectrometry (LC-MS/MS). Glucose-regulated protein 78-kd (GRP78),

cleaved caspase-3 and cyclooxygenase-1 (COX-1) protein levels were assessed

by immunofluorescence. Cytosolic phospholipase A2 (cPLA2), total COX and

prostaglandin E2 (PGE2) were measured to evaluate changes in enzyme activity.

RESULTS: Decreased cell viability was observed in TM treated cells.

Administration of TUDCA following TM treatment significantly increased cell

viability compared to TM treatment alone. Tunicamycin-induced ER stress was

confirmed by significantly increased protein levels of GRP78. A significant

increase was observed in C18-C24 CERs and caspase-3 activity, while a

significant decrease occurred in sphingosine-1-phosphate (S1P) and cPLA2

activity in cells treated with μg/ml versus controls. The decrease in cPLA2

activity was accompanied by significantly increased PUFA levels in TM treated

cells. Tauroursodeoxycholic acid treatment in conjunction with TM significantly

decreased ER stress, C18-C24 CERs, caspase 3 activity and increased S1P levels.

CONCLUSIONS: Results show the build-up of long chain CERs and PUFAs

in HEK-293 cells undergoing ER stress alongside increased apoptotic activity.

Tauroursodeoxycholic acid administration, along with TM treatment alleviated

the build-up of CERs and TM-induced apoptotic activity in kidney epithelial cells.

Keywords: Ceramide, Kidney, PUFA, Sphingolipid, Tunicamycin