Combination of zinc and selenium alleviates ochratoxin A-induced fibrosis via blocking ROS-dependent autophagy in HK-2 cells

Guannan Le; Lulu Yang; Heng Du; Lili Hou; Lei Ge; Ardache Sylia; Azhar Muhmood; Xinxiang Chen; Bo Han; Kehe Huang

doi:10.1016/j.jtemb.2021.126881

Combination of zinc and selenium alleviates ochratoxin A-induced fibrosis via blocking ROS-dependent autophagy in HK-2 cells

J Trace Elem Med Biol. 2022 Jan:69:126881. doi: 10.1016/j.jtemb.2021.126881. Epub 2021 Oct 27.

Authors

Guannan Le¹, Lulu Yang², Heng Du³, Lili Hou³, Lei Ge³, Ardache Sylia³, Azhar Muhmood³, Xinxiang Chen³, Bo Han⁴, Kehe Huang⁵

Affiliations

¹ College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, 210095, Jiangsu Province, China. Electronic address: 2018107088@njau.edu.cn.
² College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi, China.
³ College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, 210095, Jiangsu Province, China.
⁴ Department of Clinical Veterinary Medicine, College of Veterinary Medicine, China Agricultural University, Beijing, 100193, China.
⁵ College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, 210095, Jiangsu Province, China. Electronic address: khhuang@njau.edu.cn.

PMID: 34751137
DOI: 10.1016/j.jtemb.2021.126881

Abstract

Background: Ochratoxin A (OTA) is a mycotoxin produced by Aspergillus and Penicillium. The key target organ of OTA toxicity is the kidney, which has a significant impact on human health. Recently, nutrition regulation is suggested to be an effective protection against mycotoxins contamination. The current study investigated the combined protective effects of zinc and selenomethionine (SeMet) (a major component of organic selenium) on OTA-induced renal fibrosis and their potential mechanisms in human renal proximal tubule epithelial cells (HK-2 cells).

Methods: Cytotoxicity of different concentrations of OTA, zinc and SeMet on HK-2 cells was detected by cell viability, lactate dehydrogenase (LDH) and apoptotic nuclei assays. The expression of fibrosis biomarkers was detected by Real-Time PCR, western blotting and indirect immunofluorescence assays. The production of reactive oxygen species (ROS) was detected by ROS assay kit. The protein expression of autophagy biomarkers was detected by western blotting assay.

Results: Cytotoxicity was induced by OTA treatment in a dose-dependent manner, and it was attenuated by zinc or SeMet application in HK-2 cells. Zinc or SeMet application also down-regulated the expression of fibrosis biomarkers, and the combination of them displayed better effects. In addition, OTA increased intracellular ROS level and activated autophagy in a dose-dependent manner, and it was reversed by zinc and SeMet combined application. With the treatment of hydrogen peroxide (H₂O₂) or rapamycin (the specific activator of autophagy), the combined protective effects of zinc and SeMet were abolished.

Conclusions: Zinc and SeMet application alleviated OTA-induced cytotoxicity and fibrosis in HK-2 cells. Combination of them was more effective than its individual application. The present study manifest novel insight about the alleviation of OTA-induced nephrotoxicity by nutrition regulation, and had a guiding effect on the clinical supplementation of nutritional elements.

Keywords: Autophagy; Ochratoxin A; ROS; Renal fibrosis; Selenomethionine; Zinc.

MeSH terms

Antioxidants
Autophagy
Fibrosis
Humans
Hydrogen Peroxide
Ochratoxins*
Reactive Oxygen Species
Selenium* / pharmacology
Selenomethionine / pharmacology
Zinc* / pharmacology

Substances

Antioxidants
Ochratoxins
Reactive Oxygen Species
ochratoxin A
Selenomethionine
Hydrogen Peroxide
Selenium
Zinc