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. 2023 May 21;12(10):2073.
doi: 10.3390/foods12102073.

Dairy By-Products and Lactoferrin Exert Antioxidant and Antigenotoxic Activity on Intestinal and Hepatic Cells

Inés Abad  1   2 , Julien Vignard  3 , Catherine Bouchenot  3 , Dimitra Graikini  1   2 , Laura Grasa  2   4 , María Dolores Pérez  1   2 , Gladys Mirey  3 , Lourdes Sánchez  1   2
Affiliations

Dairy By-Products and Lactoferrin Exert Antioxidant and Antigenotoxic Activity on Intestinal and Hepatic Cells

Inés Abad et al. Foods. .

Abstract

The dairy industry generates a large volume of by-products containing bioactive compounds that may have added value. The aim of this study was to evaluate the antioxidant and antigenotoxic effects of milk-derived products, such as whey, buttermilk, and lactoferrin, in two human cell lines: Caco-2 as an intestinal barrier model and HepG2 as a hepatic cell line. First, the protective effect of dairy samples against the oxidative stress caused by menadione was analyzed. All these dairy fractions significantly reversed the oxidative stress, with the non-washed buttermilk fraction presenting the greatest antioxidant effect for Caco-2 cells and lactoferrin as the best antioxidant for HepG2 cells. At concentrations that did not impact cell viability, we found that the dairy sample with the highest antigenotoxic power against menadione, in both cell lines, was lactoferrin at the lowest concentration. Additionally, dairy by-products maintained their activity in a coculture of Caco-2 and HepG2, mimicking the intestinal-liver axis. This result suggests that the compounds responsible for the antioxidant activity could cross the Caco-2 barrier and reach HepG2 cells on the basal side, exerting their function on them. In conclusion, our results show that dairy by-products have antioxidant and antigenotoxic activities, which would allow revaluing their use in food specialties.

Keywords: Caco-2 cells; DNA damage; HepG2 cells; bioavailability; buttermilk; lactoferrin; oxidative stress; whey.

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Conflict of interest statement

There are no known conflicts of interest associated with any of the authors participating in this publication, and there has been no significant financial support for this work that could have influenced its outcome.

Figures

Figure 1
Figure 1
Viability of (A) Caco-2 and (B) HepG2 cells after 24 h of incubation with dairy fractions. NT: non-treated cells. Cali: positive control of cytotoxicity consisting of cells treated with 5 µM calicheamicin. LF: lactoferrin. WBM: washed buttermilk. NWBM: non-washed buttermilk. The viability is expressed in percentages with respect to NT. The values represent the mean ± standard deviation of two replicates in three independent experiments (n = 6). * p < 0.05, ** p< 0.01, *** p < 0.001, **** p < 0.0001, compared with NT.
Figure 2
Figure 2
Effect of dairy fractions on oxidative stress caused by menadione on (A) Caco-2 and (B) HepG2 cells. NT: non-treated cells. MEN: positive control of oxidative stress, cells treated with 50 µM menadione for 1 h. NAC + MEN: cells treated for 1 h with 1 mM NAC as a control of antioxidant effect before menadione treatment. LF: lactoferrin. WBM: washed buttermilk. NWBM: non-washed buttermilk. The results were normalized to 1 for the untreated condition. The values represent the intensity of 150–200 cells counted in each experiment, on three independent experiments. The bars represent the means of all counts. **** p < 0.0001, compared with MEN.
Figure 3
Figure 3
Effect of dairy fractions on genotoxicity caused by menadione on (A) Caco-2 and (B) HepG2 cells. NT: non-treated cells. MEN: positive control of genotoxicity, cells treated with 50 µM menadione for 1 h. NAC + MEN: cells treated with 1 mM NAC as a control of antigenotoxic effect before treatment with 50 µM menadione for 1 h. LF: lactoferrin. WBM: washed buttermilk. NWBM: non-washed buttermilk. The results were normalized to 1 for the untreated condition. The values represent the intensity of 150–200 cells counted in each experiment, on three independent experiments. The bars represent the means of all counts. **** p < 0.0001, compared with MEN.
Figure 4
Figure 4
Effect of dairy fractions on oxidative stress (A,B) and genotoxicity (C,D) caused by menadione on Caco-2 cells (A,C) and HepG2 cells (B,D) co-cultured in standard transwell inserts. NT: non-treated cells. MEN: positive control of oxidative stress and genotoxicity, cells treated with 50 µM menadione. NAC + MEN: cells treated with 1 mM NAC as a control of antioxidant and antigenotoxic effects before treatment with 50 µM menadione. LF: lactoferrin. WBM: washed buttermilk. NWBM: non-washed buttermilk. The results were normalized to 1 for the untreated condition. The values represent the intensity of 150–200 cells counted in each experiment, on three independent experiments. The bars represent the means of all counts. **** p < 0.0001, compared with MEN.
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References

    1. Svanborg S., Johansen A.-G., Abrahamsen R.K., Skeie S.B. The composition and functional properties of whey protein concentrates produced from buttermilk are comparable with those of whey protein concentrates produced from skimmed milk. J. Dairy Sci. 2015;98:5829–5840. doi: 10.3168/jds.2014-9039. - DOI - PubMed
    1. Lin K.-L., Wang P.-W., Chuang J.-H., Lin H.-Y., Chen S.-D., Chuang Y.-C., Huang S.-T., Tiao M.-M., Chen J.-B., Huang P.-H., et al. Resveratrol provides neuroprotective effects through modulation of mitochondrial dynamics and ERK1/2 regulated autophagy. Free. Radic. Res. 2018;52:1371–1386. doi: 10.1080/10715762.2018.1489128. - DOI - PubMed
    1. Cichosz G., Czeczot H., Bielecka M. The anticarcinogenic potential of milk fat. Ann. Agric. Environ. Med. 2020;27:512–518. doi: 10.26444/aaem/116095. - DOI - PubMed
    1. Domingos J.M., Puccio S., Martinez G.A., Amaral N., Reis M.A., Bandini S., Fava F., Bertin L. Cheese whey integrated valorisation: Production, concentration and exploitation of carboxylic acids for the production of polyhydroxyalkanoates by a fed-batch culture. Chem. Eng. J. 2018;336:47–53. doi: 10.1016/j.cej.2017.11.024. - DOI
    1. Eurostat Statistics Explained. 2021. Milk and Milk Product Statistics. [(accessed on 17 February 2023)]. Available online: https://ec.europa.eu/eurostat/statistics-explained/index.php?title=Milk_....

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