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Asian-Australas J Anim Sci > Accepted Articles
DOI: https://doi.org/10.5713/ajas.20.0349    [Accepted] Published online August 24, 2020.
Protective Effects of 5-Aminolevulinic Acid on Heat Stress in Bovine Mammary Epithelial Cells
Md Aminul Islam1  , Yoko Noguchi2  , Shin Taniguchi2  , Shinichi Yonekura1,2,* 
1Graduate School of Medicine, Science and Technology, 3 Department of Biomolecular Innovation, Institute for Biomedical Sciences, Interdisciplinary Cluster for Cutting Edge Research, Shinshu University, Kamiina, Nagano 399-4598, Japan
2Neopharma Japan Co., Ltd. Tokyo 102-0071, Japan
Correspondence:  Shinichi Yonekura, Tel: +81-265771443, Fax: +81-265771443, Email: yonekura@shinshu-u.ac.jp
Received: 18 May 2020   • Revised: 27 June 2020   • Accepted: 27 July 2020
Abstract
Objective
Cells have increased susceptibility to activation of apoptosis when suffering heat stress (HS). An effective supplementation strategy to mimic heat-induced apoptosis of bovine mammary epithelial cells (MECs) is necessary to maintain optimal milk production. This study aimed to investigate possible protective effects of the anti-apoptotic activity of 5-aminolevulinic acid (5-ALA) against HS-induced damage of bovine MECs.
Methods
Bovine MECs were pretreated with or without 5-ALA at concentrations of 10, 100, and 500 μM for 24 h followed by HS (42.5°C for 24 h and 48 h). Cell viability was measured with MTT assays. Real-time qPCR and Western blotting were used to explore the regulation of genes associated with apoptosis, oxidative stress, and endoplasmic reticulum (ER) stress genes.
Results
We found that 5-ALA induces cytoprotection via inhibition of apoptosis markers after HS-induced damage. Pretreatment of bovine MECs with 5-ALA resulted in dramatic upregulation of mRNA for nuclear factor erythroid-derived 2-like factor 2 (NRF2), heme oxygenase-1 (HO-1), and NAD(P)H quinone oxidoreductase 1 (NQO1), all of which are antioxidant stress genes. Moreover, 5-ALA pretreatment significantly suppressed HS-induced ER stress-associated markers, glucose-regulated protein 78 (GRP78), and C/EBP homologous protein (CHOP) expression levels.
Conclusion
5-ALA has the ability to ameliorate the ER stress in heat stressed bovine mammary epithelial cell via enhancing the expression of antioxidant gene.
Keywords: Heat Stress; 5-Aminolevulinic Acid; Bovine Mammary Epithelial Cells; ER Stress; Oxidative Stress


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