Effect and mechanism of Juluo decoction combined with Ruhe powder in treatment of rat model of hyperplasia of mammary glands

Objective To investigate the effect and mechanism of Juluo Decoction combined with Ruhe Powder in the treatment of rat model of hyperplasia of mammary glands. Methods A total of 40 specific pathogen-free female Sprague-Dawley rats, aged 7-8 weeks, were randomly divided into control group (group A), hyperplasia of mammary glands group (group B), Rupixiao treatment group (group C), and Juluo Decoction+Ruhe Powder treatment group (group D), with 10 rats in each group. The rats in groups B, C, and D were given intramuscular injection of estradiol benzoate and progesterone to establish a model of hyperplasia of mammary glands, and those in group A were given an equal volume of saline. After successful modeling, the rats in group D were given Juluo Decoction by gavage, with the second and third pairs of mammary glands wrapped with elastic bands containing Ruhe Powder extract for 30 days; the rats in group C were given Rupixiao solution by gavage, while those in groups A and B were given an equal volume of normal saline by gavage, and for the rats in groups A, B, and C, the second and third pairs of mammary glands were wrapped with elastic bands for 30 days. After the end of drug treatment, nipple height and breast diameter were measured for the second pair of mammary glands in each group; ELISA was used to measure the serum levels of estradiol (E2), progesterone, follicle-stimulating hormone (FSH), and luteinizing hormone (LH); HE staining and Masson staining were used to observe the pathological changes of mammary gland tissue in each group; Western blotting was used to measure the relative expression levels of proteins associated with the phosphatidylinositol 3-kinase (PI3K)/serine-threonine kinase (AKT)/B-cell lymphoma-2 (Bcl-2) signaling pathway and the extracellular signal-regulated kinase (ERK) signaling pathway in mammary gland tissue; HE staining was used to observe the histomorphology of the liver and the kidney. Results Compared with group B, groups C and D had significant reductions in nipple height and breast diameter, and group D had a significantly lower nipple height than group C (F=104.10,172.70,t=2.54-12.24,P<0.05). ELISA showed that compared with group B, groups C and D had significant reductions in the serum levels of E2 and FSH and significant increases in the serum levels of progesterone and LH, and group D had significantly higher serum levels of progesterone and LH than group C (F=16.46-149.10,t=4.53-15.25,P<0.05). HE staining and Masson staining showed that compared with group B, groups C and D had significant reductions in the glands in mammary gland tissue, significant alleviation of duct dilation and duct epithelial cell proliferation, and significant reductions in duct secretions and collagen deposition, and compared with group C, group D had significantly greater improvements in these pathological features. Western blotting showed that compared with group B, groups C and D had significant reductions in the ratios of p-AKT/AKT, Bcl-2/Bax, and p-ERK/ERK and the relative expression level of ERα in mammary gland tissue, and compared with group C, group D had a significantly lower relative expression level of ERα in mammary gland tissue (F=21.37-205.50,t=4.94-30.51,P<0.05). HE staining showed that there were no obvious abnormalities in the histomorphology of the liver and the kidney. Conclusion Juluo Decoction combined with Ruhe Powder can alleviate breast enlargement, sex hormone disorder, and the pathological changes of breast tissue in a rat model of hyperplasia of mammary glands, possibly by inhibiting the activation of the PI3K/AKT/Bcl-2 and ERK signaling pathways.