Intervention Effect and Mechanism of Compound Herba Gueldenstaedtiae in a Mouse Model of Breast Hyperplasia

Abstract

BACKGROUND: Breast hyperplasia is a common benign breast disease mainly caused by endocrine disorders, manifested as abnormal hyperplasia of breast tissue. In recent years, traditional Chinese medicine compounds and probiotics have shown good potential in regulating the endocrine system and improving the intestinal microecology, providing new ideas for the treatment of breast hyperplasia. OBJECTIVE: To explore the effects and mechanisms of traditional Chinese medicine compounds and fermented probiotic compounds on breast hyperplasia in mice, providing new theoretical and experimental bases for the clinical treatment and prevention of breast hyperplasia. METHODS: (1) Network pharmacology tools were used to predict the anti-breast-hyperplasia activity of Herba Gueldenstaedtiae (Euphorbia humifusa), as well as its potential targets and signaling pathways. The databases included: TCMSP, OMIM, GeneCards database, UniProt website, Venny2.1.0 website, Metascape, HERB website, and STRING database, all of which are open-access databases. Network pharmacology can predict and screen key information such as the targets corresponding to the active ingredients of traditional Chinese medicine, disease targets, and action pathways through network analysis and computer-system analysis. Therefore, it has been increasingly widely used in the research of traditional Chinese medicine. (2) A breast hyperplasia model was induced in mice by injecting estrogen and progesterone. Mice in the normal blank group were injected intraperitoneally with normal saline every day. Mice in the model group and drug-administration groups were injected intraperitoneally with estradiol benzoate injection at a concentration of 0.5 mg/kg every day for 25 days. From the 26th day, the injection of estradiol benzoate injection was stopped. Mice in the normal blank group were injected intramuscularly with normal saline every day, and mice in the model group and drug-administration groups were injected intramuscularly with progesterone injection at a concentration of 5 mg/kg for 5 days. After the model was established, each group was given drugs respectively. The normal blank group and the model group were gavaged with 0.2 mL/d of normal saline; the positive blank group (Xiaozheng Pill group) was gavaged with an aqueous solution of Xiaozheng Pill at 0.9 mg/g; the low-, medium- and high-dose groups of Compound Herba Gueldenstaedtiae were gavaged with an aqueous solution of the compound medicine at 0.75, 1.5, and 3.0 mg/(g·d) respectively; the low-, medium- and high-dose groups of traditional Chinese medicine-bacteria fermentation were gavaged with an aqueous solution of the compound medicine at 0.75, 1.5, and 3.0 mg/(g·d) respectively. The administration was continuous for 30 days. RESULTS AND CONCLUSION: (1) The results of network pharmacology research showed that the Compound Herba Gueldenstaedtiae (Euphorbia humifusa) contained 46 active ingredients, which were related to 1 213 potential targets. After comparison with 588 known breast-hyperplasia targets, it was speculated that 50 of these targets might be related to the direct effect of the compound on breast hyperplasia. (2) After drug intervention, there was no significant change in the high-dose group of Compound Herba Gueldenstaedtiae compared with the normal blank group. The liver indicators of the other intervention groups all significantly decreased (P < 0.05). (3) In terms of kidney and uterine indicators, the medium-dose group of Compound Herba Gueldenstaedtiae decreased significantly compared with the normal blank group (P < 0.05). In terms of the uterine index, the model group increased significantly compared with the normal blank group (P < 0.01). (4) After 1-month drug treatment, the number of lobules and acini in the breast tissue of the Xiaozheng Pill group, the low, medium, and high-dose group of Compound Herba Gueldenstaedtiae, the low, medium, and high-dose groups of traditional Chinese medicine-bacteria fermentation decreased, and the duct openings narrowed. With the increase of drug dose, diffuse hyperplasia of breast tissue was significantly improved. (5) The ELISA results showed that compared with the model group, the estrogen level was lower in the medium-dose group of traditional Chinese medicine-bacteria fermentation after the intervention (P < 0.05). In addition, the follicle-stimulating hormone level in the low-dose group of Compound Herba Gueldenstaedtiae was lower than that of the model group (P < 0.05). (6) The intervention in the mouse model led to changes in the abundance of short chain fatty acids and intestinal flora in all groups. To conclude, the Compound Herba Gueldenstaedtiae and its probiotic fermentation products significantly improved mammary gland hyperplasia in mice by regulating hormone levels, improving the structure of the gut microbiota, and increasing the content of short-chain fatty acids, providing new ideas and potential sources of drugs for the treatment of breast hyperplasia.

1. Introduction

Breast hyperplasia is mainly a benign disorder of breast structure caused by endocrine disorders, which is closely related to the changes in the levels of hormones such as estrogen and progesterone[1-3]. Notably, breast hyperplasia is recognized as an important risk factor for breast cancer. Therefore, early intervention for breast hyperplasia is crucial to prevent its progression to breast cancer, which may otherwise affect the physical and mental health of patients. This disease is particularly common among women aged 35–49 years[4].

Breast hyperplasia is not only caused by endocrine disorders. Research shows that when the gut microbiota is severely imbalanced, it can also affect the body’s endocrine system, especially the breast tissue of women[5,6]. Moreover, an increasing number of studies indicate that gut microbiota dysbiosis may also affect the body’s endocr...