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. 2019 Sep 23;55(10):625.
doi: 10.3390/medicina55100625.

Melatonin Action on the Activity of Phagocytes from the Colostrum of Obese Women

Tassiane C Morais  1   2 , Adenilda C Honorio-França  3 , Mahmi Fujimori  4 , Ocilma B de Quental  5   6 , Rafael S Pessoa  7 , Eduardo L França  8 , Luiz C de Abreu  9   10   11   12
Affiliations

Melatonin Action on the Activity of Phagocytes from the Colostrum of Obese Women

Tassiane C Morais et al. Medicina (Kaunas). .

Abstract

Background and objectives: Breastfeeding promotion is an important public health strategy for counter-balancing the negative effects of maternal overweight and obesity. Colostrum contains melatonin, which can attenuate the impacts of excessive maternal weight and boost the infant's immune system. Therefore, the objective of this study was to analyze the effects of melatonin on mononuclear (MN) phagocytes from the colostrum of women with pre-gestational obesity. Materials and Methods: Colostrum samples were collected postpartum from 100 women at a public hospital in São Paulo, Brazil. The donors were divided into two groups: the control group and the high body mass index (BMI) group. Melatonin levels in the colostrum were determined by an ELISA Kit, and the functional activity of MN cells was assessed using the phagocytosis assay by flow cytometry, and reactive oxygen species (ROS), intracellular calcium, and apoptosis were assessed by fluorimetry using a microplate reader.

Results: The colostrum of mothers with pre-gestational high BMI exhibited higher melatonin levels (p < 0.05) and lower phagocytosis (p < 0.05) and ROS release (p < 0.05). Superoxide release was similar between the normal and high BMI groups (p > 0.05). Intracellular calcium release and apoptosis were also higher in the high BMI group (p < 0.05). Melatonin levels likely increased the phagocytosis rate and reduced intracellular calcium release and the apoptosis index (p < 0.05).

Conclusions: The results suggest that melatonin is a possible mechanism for maternal-infant protection against obesity and restores the functional activity of colostrum phagocytes in obese mothers.

Keywords: body mass index; breastmilk; colostrum; melatonin; obesity; oxidative stress; phagocytes.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Melatonin (pg/mL) levels in the colostrum of mothers with normal or high BMI pregestational. * Assessed by ANOVA and Tukey’s test. * p < 0.05 Statistical difference between normal BMI and high BMI group. BMI, Body Mass Index; ANOVA, A D’Agostino normality test and variance analysis.
Figure 2
Figure 2
High BMI effects on colostrum mononuclear (MN) phagocytes. (a) Phagocytosis rate (%) in colostrum MN cells, from normal and obese women, incubated or not with melatonin. (b) Fluorescence microscopy image of pH Rodo® Green zymosan phagocytosis by MN cells, after 2 h of incubation (scale: 5 μm, 100 x—panel). The results are expressed as the mean ± standard deviation (SD). * statistical difference (p < 0.05) between control cells (199 medium) and cells incubated with zymosan (Zy) or zymosan and melatonin (Zy + MLT) within a same group. #Statistical difference among groups with the same treatment and sample (p < 0.05).
Figure 3
Figure 3
Reactive oxygen species release by colostrum phagocytes. (a) Fluorescence intensity of DHR123. (b) Superoxide release (nmol/mL). Colostrum phagocytes were incubated with medium 199 or Phosphate Buffer Solution (PBS), melatonin (100 ng/mL) and/or zymosan. DRH123 was analyzed using Fluoroskan Ascent FL® Microplate. The results are expressed as the mean ± standard deviation (SD). * Statistical difference (p < 0.05) between control cells (PBS or 199 medium) and cells incubated with zymosan (Zy) or zymosan and melatonin (Zy + MLT), within the same group; #Statistical difference among groups with the same treatment and sample (p < 0.05).
Figure 4
Figure 4
Intracellular calcium release (a) and apoptosis rates (b) by colostral phagocytes. The phagocytes were incubated with 199 medium, Zymosan, and melatonin (100 ng/mL). The intracellular calcium release was performed with Fluo-3AM (n = 10). The apoptosis assay was performed with Annexin V-FITC staining. The results are presented as the mean ± standard deviation (SD). * p < 0.05, * statistical difference (p < 0.05) between the control cells (199 medium) and cells incubated with zymosan (Zy) or zymosan and melatonin (Zy + MLT) within the same group. +Statistical difference (p < 0.05) between cells incubated with zymosan, treated or not with melatonin, within the same group. # The statistical difference among groups with the same treatment and the sample.
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