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Two major bovine milk whey proteins induce distinct responses in IEC-6 intestinal cells

Published online by Cambridge University Press:  23 September 2024

Mizuho Inagaki ,
Midori Nohara ,
Chihiro Kobayashi ,
Mayuko Fukuoka ,
Xijier ,
Keisuke Matsumura ,
Kiyomi Ohashi ,
Haru Enomoto ,
Tomio Yabe  and
Yoshihiro Kanamaru
Mizuho Inagaki*
Affiliation:
Department of Applied Life Science, Faculty of Applied Biological Sciences, Gifu University, Gifu, Japan
Midori Nohara
Affiliation:
Department of Applied Life Science, Faculty of Applied Biological Sciences, Gifu University, Gifu, Japan
Chihiro Kobayashi
Affiliation:
Department of Applied Life Science, Faculty of Applied Biological Sciences, Gifu University, Gifu, Japan
Mayuko Fukuoka
Affiliation:
Department of Applied Life Science, Faculty of Applied Biological Sciences, Gifu University, Gifu, Japan
Xijier
Affiliation:
Department of Applied Life Science, Faculty of Applied Biological Sciences, Gifu University, Gifu, Japan
Keisuke Matsumura
Affiliation:
Department of Applied Life Science, Faculty of Applied Biological Sciences, Gifu University, Gifu, Japan
Kiyomi Ohashi
Affiliation:
Department of Applied Life Science, Faculty of Applied Biological Sciences, Gifu University, Gifu, Japan
Haru Enomoto
Affiliation:
Department of Applied Life Science, Faculty of Applied Biological Sciences, Gifu University, Gifu, Japan
Tomio Yabe
Affiliation:
Department of Applied Life Science, Faculty of Applied Biological Sciences, Gifu University, Gifu, Japan Institute for Glyco-core Research (iGCORE), Gifu University, Gifu, 501-1193, Japan Preemptive Food Research Center (PFRC), Gifu University Institute for Advanced Study, Gifu, 501-1193, Japan
Yoshihiro Kanamaru
Affiliation:
Department of Applied Life Science, Faculty of Applied Biological Sciences, Gifu University, Gifu, Japan
*
Corresponding author: Mizuho Inagaki; Email: [email protected]
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Abstract

α-Lactalbumin (α-LA) and β-lactoglobulin (β-LG) are major whey proteins in bovine milk. We studied the effects of these molecules on the intestinal cell response by comparing the native form with the denatured form containing oligomers obtained by treatment with 2,2,2-trifluoroethanol (TFE). We previously reported that proteins in native and TFE-treated forms exhibited cell growth stimulation and cytotoxicity, respectively, in undifferentiated rat crypt IEC-6 and human colon Caco-2 cells. However, neither whey protein showed cytotoxicity even in the TFE-treated form in differentiated Caco-2 cells. Only undifferentiated immature intestinal cells can distinguish between these native and denatured proteins. Moreover, α-LA and β-LG exhibited different oligomer formation characteristics during the TFE treatment. In the present study, we compared the effects of native and TFE-treated whey proteins on IEC-6 cells in more detail. The native forms of both whey proteins exhibited cell proliferative effects in a concentration-dependent manner. For the TFE-treated forms, α-LA showed rapid and potent cytotoxicity, whereas β-LG altered cell responses depending on its concentration and exposure time; lower concentration/shorter exposure and higher concentration/longer exposure induced cell growth stimulation and cytotoxicity, respectively. Pre-treatment of the cell membrane with cholesterol suppressed the effects on the cell response only in TFE-treated β-LG (TFE-β-LG). In a preliminary examination using inhibitors of signal transduction, TFE-treated α-LA acted on the intrinsic apoptosis pathway via Bcl-2-associated X and p53, whereas the action of TFE-LG did not require this pathway. Tyrosine phosphorylation is necessary for the cell proliferation effect of both native whey proteins; however, native α-LA, but not native β-LG, also required activation of the pathway with selective epidermal growth factor receptor tyrosine kinase and Janus kinase 2/3. In summary, the two major bovine milk whey proteins induced similar yet discrete responses in undifferentiated intestinal cells. Even when oligomers are formed, β-LG may be much less hazardous to immature intestinal cells than α-LA.

Keywords

α-Lactalbuminβ-lactoglobulinapoptosiscell proliferationmisfolding
Type
Research Article
Information
Journal of Dairy Research , Volume 91 , Issue 2 , May 2024 , pp. 239 - 245
Copyright
Copyright © The Author(s), 2024. Published by Cambridge University Press on behalf of Hannah Dairy Research Foundation

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