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. 2022 Jun 18:2022:5762847.
doi: 10.1155/2022/5762847. eCollection 2022.

Medicinal Mushroom Leucocalocybe mongolica Imai Extracts Improve Mammary Gland Differentiation in Lactating Rats via Regulating Protein Expression

Asmaa Hussein Zaki  1   2   3 , Bao Haiying  1   2 , Li Zhijun  1   2
Affiliations

Medicinal Mushroom Leucocalocybe mongolica Imai Extracts Improve Mammary Gland Differentiation in Lactating Rats via Regulating Protein Expression

Asmaa Hussein Zaki et al. Evid Based Complement Alternat Med. .

Abstract

Leucocalocybe mongolica is a known medicinal mushroom in China. It possesses many biological activities. This study investigated the effect of L. mongolica petroleum ether and water extracts (200, 500, and 1,000 mg/kg BW) on mammary gland differentiation during lactation. However, prolactin, growth hormone, progesterone, and estrogen levels were determined in serum by ELISA assay. Immunofluorescence, western blot, and real-time PCR were utilized to evaluate the expression levels of β-casein, α-Lactalbumin, prolactin receptor, progesterone receptor, and STAT-5a. The immunohistochemistry staining was used to detect the presence of steroid receptors. The results showed that petroleum ether and water extracts increased milk yield and milk content of calcium, total fat, total carbohydrate, and total protein. Prolactin and growth hormone levels were significantly upregulated in all treated groups compared with the control group. In contrast, progesterone and estrogen were downregulated. The high doses of petroleum ether and water extracts increased the expression levels of β-Cas, α-Lactalb, PRLR, PR, and STAT-5a. The observation of histological sections showed that the extracts induced higher mammary gland differentiation than the control group. This study is the first to use mushrooms as nutritional supplements to improve milk production and mammary gland differentiation during lactation.

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Conflict of interest statement

The authors declare that there are no conflicts of interest.

Figures

Figure 1
Figure 1
Effect of L. mongolica extracts on cell viability. MCF10 A cell lines were treated with L. mongolica petroleum ether extract (PE), water extract (WE), metoclopramide (MET), and CON control (50, 100, and 200 µg/mL) for 48 h. The cell viability was measured by CCK8. The values are represented as the mean ± SD standard deviation of three independent experiments. Compared with the CON group,  ∗P < 0.05 and  ∗ ∗P < 0.01. The normality and homoscedasticity of the data were verified using the Shapiro–Wilk and Levene's tests, respectively.
Figure 2
Figure 2
Effects of L. mongolica extracts on milk production for 18 days. The values are represented in the chart as mean ± SD standard deviation. Compared with the CON group,  ∗P < 0.05 and  ∗ ∗P < 0.01. The normality of the data within the groups was verified using the Shapiro–Wilk test, and the homoscedasticity was checked by Levene's tests.
Figure 3
Figure 3
Representative images of hematoxylin and eosin-stained sections of mammary alveolar tissue from the lactating with H&E (400×). The arrows refer to the active alveoli with lobules;  ∗to the inter lobules duct; f refers to internal lobular connective tissue; and d refers to the adipose tissue.
Figure 4
Figure 4
Representative immunofluorescence images of rat mammary alveoli and positive average absorbance indicating α-Lactalb expression. The alveolar cell's cytoplasms and apical lumens were positive for α-Lactalb (green), while their nuclei were negative (blue, DAPI+). The mean ± SD standard deviation is used to express the values. (Compared with the CON group,  ∗P < 0.05,  ∗ ∗P < 0.01, and  #P < 0.05 compared with the MET group.
Figure 5
Figure 5
The immunohistochemistry staining (200x) and percentages of alveolar cells positive for ER-α in the rat mammary tissue. The bar chart expresses the mean values ± SD.  ∗P < 0.05, compared with the CON group, and  #P < 0.05 compared with the MET group.
Figure 6
Figure 6
mRNA expression of β-Cas (a), α-Lactalb (b), PRLR (c), and STAT-5 (d) by real-time PCR. The bars represent the mean values ± SD. Compared with the CON group,  ∗P < 0.05,  ∗ ∗P < 0.01, and  #P < 0.05 compared with the MET group.
Figure 7
Figure 7
The expression levels of PR (a), PRLR (b), STAT-5a (c), α-Lactalb (d), and β-Cas (e) proteins in the mammary tissues and typical images of western blots. The data are expressed per µg of tissue, and the bars indicate the mean values ± SD. (Compared with the CON group,  ∗P < 0.05,  ∗ ∗P < 0.01, and  #P < 0.05 compared with the MET group.
Figure 8
Figure 8
The mechanism by which L. mongolica improves mammary gland proliferation and increases milk protein expression.
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