ABSTRACT
This study aimed to investigate the mechanism of Xihuang Pills in improving hyperplasia of mammary gland(HMG) in rats based on urine metabolomics using ultra-performance liquid chromatography-quadrupole-Orbitrap mass spectrometry(UPLC-Q-Orbitrap-MS). The HMG rat model was established by intramuscular injection of estradiol benzoate solution(0.5 mg·kg~(-1), 25 days) followed by progesterone injection(5 mg·kg~(-1), 5 days). UPLC-Q-Orbitrap-MS technology was used to establish the endogenous small-molecule metabolic profiles in urine samples of rats in the blank group, the HMG model group, and Xihuang Pills group. Multivariate statistical analysis was performed for pattern recognition, t test and variable importance in the projection(VIP) were used to screen potential biomarkers. The significantly changed differential metabolites were identified using the online database Human Metabolome Database(HMDB). Metabolic pathway enrichment analysis was conducted using the MetaboAnalyst 5.0 database. The results showed that 90 differential metabolites with significant changes(P<0.05) were identified between the blank group and the HMG model group using the HMDB. Among them, 48 metabolites significantly reverted(P<0.05) after administration of Xihuang Pills, which may be related to the regulatory effect of Xihuang Pills. Thirteen metabolic pathways significantly associated with HMG were identified when the differential metabolites were imported into the MetaboAnalyst 5.0 database, and Xihuang Pills could modulate seven of these pathways. These metabolic pathways mainly involved histidine metabolism, arginine and proline metabolism, β-alanine metabolism, glycine, serine and threonine metabolism, tryptophan metabolism, pyrimidine metabolism, and amino sugar and nucleotide sugar metabolism. This study utilized UPLC-Q-Orbitrap-MS and urine metabolomics technology to analyze the mechanism of Xihuang Pills in improving HMG, laying the foundation for further in-depth research.
Subject(s)
Humans , Rats , Animals , Chromatography, High Pressure Liquid/methods , Hyperplasia , Metabolomics/methods , Metabolome , Biomarkers/urineABSTRACT
Objective:The SD rat model of hyperplasia of mammary gland（HMG） and the ultrahigh-performance liquid chromatography and mass spectrometry (UHPLC-LTQ-Orbitrap MS) technology were used to explore the pharmacological material basis of Shuangjin Sanjie granules (SJSJG) for the treatment on HMG.Method:SD rat models of HMG were administered in groups, and the nipple height and the diameter were measured; the levels of estradiol (E2), progesterone (P) and prolactin (PRL) in serum were detected, pathological examination was conducted for the hyperplasia of breast tissue. Histochemical methods were used to detect the expressions of estrogen receptor α (ERα), androgen receptor (AR), progesterone receptor (PR), tumor necrosis factor-α (TNF-α) proteins. Finally, UHPLC-LTQ-Orbitrap MS technology was used to detect the main chemical constituents of SJSJG, and the pharmacodynamic substance basis was analyzed based on the pharmacological effect.Result:The results of animal experiments showed that compared with the normal group, nipple height and diameter of the model group increased remarkably (P<0.01), serum E2 significantly increased (P<0.01). Pathological examination showed abnormal hyperplasia of breast tissue， expressions of ERα, AR, PR and TNF-α increased， compared with the model group, the nipple height and diameter of the SJSJG group decreased remarkably (P<0.01), serum E2 was decreased significantly (P<0.01)， pathological examination showed weakened abnormal hyperplasia of breast tissue, ERα, AR, PR and TNF-α protein expressions were significant decreased (P<0.01). The results of basic material study showed that 85 chemical components were identified from SJSJG, including 16 alkaloids, 7 flavonoids, 15 terpenes, 9 phenolic acid compounds, 3 coumarin compounds, 10 esters and lactone compounds, 7 fatty acids compounds, 4 amino acids compounds, and 14 other types of ingredients， among them, alkaloids and terpenoids chemical drug substances were closely related.Conclusion:SJSJG can effectively improve the condition of breast hyperplasia, and its medicinal substance basis may include saikosaponin A, Saikosaponin D, verticinone, peimine.
ABSTRACT
Objective:To determine the inhibitory effect of Rupixiao ( RPX) granule on mammary gland hyperplasia ( HMG) in rabbits and explore the possible mechanism to provide reference for clinical medication. Methods:Rabbit model of mammary hyperpla-sia was established by estradiol benzoate and progesterone. Xiaoyao pills and tamoxifen were used as the positive control, and the RPX granule group was respectively at low, medium and high dose (0. 525,1. 05,2. 1 g·kg-1). Each group was with intragastric adminis-tration for 30 days. The levels of E2 , PROG, FSH, LH and PRL in serum and the expression of VEGF in the homogenate tissue were determined by an Elisa method. The breast tissue of rabbits in each group was withdrawn to observe the structure changes after Hema-toxylin-eosin staining ( HE) . Results:Ovarian preservation-benzoic acid, estradiol combined with progesterone could be used to estab-lish mammary gland hyperplasia in rabbits. RPX granule could significantly decrease the serum levels of E2 and PRL (P<0. 01)and increase that of PROG (P<0. 01 or P<0. 001). The results of pathological section showed that the breast structure in RPX granule high dose group could be restored to the level of the blank control group, which showed its therapeutic effect on HMG was better than that of tamoxifen and Xiaoyao pills. The expression of VEGF in each treatment group significantly decreased compared with that in the model group by the Elisa reagent (P<0. 001). Conclusion:RPX granule has good efficacy in HMG rabbits. The inhibitory effects of RPX granule on HMG in rabbits maybe related to the inhibition of VEGF in rabbit breast tissues, and the inhibition of RPX granule was similar to that of tamoxifen and Xiaoyao pills.
ABSTRACT
Objective To observe the effect of Rule Granules on the breast tissue pathology,ovarian hormone and receptor expression of mammary glands hyperplasia rats; To explore its mechanism of action. Methods The model of mammary gland hyperplasia was established by intramuscular injection of estradiol benzoate for 20 days and intramuscular injection of progesterone for 5 d. 66 female SD rats were randomly divided into 6 groups, including blank group, model group, acupuncture group, TCM high-, medium- and low-dose groups, with 11 rats in each group. Acupuncture group chose group A (Wuyi, Hegu, on both sides, Tanzhong) and group B (Tianzong, Ganshu, Zusanli, on both sides), and the two groups were used alternately, one group per day. TCM groups received 1.1 g/mL, 0.55 g/mL,and 0.11 g/mL dose of Rule Granules for gavage,once a day.Abdominal aorta blood was collected after continuous treatment of 30 d. The contents of serum E2and P were detected through ELISA method, at the same time, the second left side of the breast was removed. The changes of the breast tissue was observed under light microscope, and the ER and PR positive expression was observed by SABC immunohistochemical method. Results Compared with the blank group, hyperplasia of mammary gland in the model group rats was more obvious, and serum E2, ER and PR positive expression of integral increased significantly (P<0.01), but the content of P decreased significantly (P<0.01); compared with the model group, hyperplasia of mammary gland was improved in all treatment groups, and serum E2, ER and PR positive expression of integral decreased significantly (P<0.05); the content of P in acupuncture group increased significantly (P<0.01), and the acupunture group and TCM low-dose group were the most obvious. Conclusion Rule Granules can effectively improve hyperplasia of mammary gland organization structure in rats caused by exogenous estrogen and progesterone, with TCM low-dose group and acupuncture group having the best efficacy. Its mechanism is related to reducing the contents of serum E2and ER, and PR positive expression in breast tissue, and increasing the content of serum P.
ABSTRACT
Objective To observe the effect of Rule Granules on the breast tissue pathology,ovarian hormone and receptor expression of mammary glands hyperplasia rats; To explore its mechanism of action. Methods The model of mammary gland hyperplasia was established by intramuscular injection of estradiol benzoate for 20 days and intramuscular injection of progesterone for 5 d. 66 female SD rats were randomly divided into 6 groups, including blank group, model group, acupuncture group, TCM high-, medium- and low-dose groups, with 11 rats in each group. Acupuncture group chose group A (Wuyi, Hegu, on both sides, Tanzhong) and group B (Tianzong, Ganshu, Zusanli, on both sides), and the two groups were used alternately, one group per day. TCM groups received 1.1 g/mL, 0.55 g/mL,and 0.11 g/mL dose of Rule Granules for gavage,once a day.Abdominal aorta blood was collected after continuous treatment of 30 d. The contents of serum E2and P were detected through ELISA method, at the same time, the second left side of the breast was removed. The changes of the breast tissue was observed under light microscope, and the ER and PR positive expression was observed by SABC immunohistochemical method. Results Compared with the blank group, hyperplasia of mammary gland in the model group rats was more obvious, and serum E2, ER and PR positive expression of integral increased significantly (P<0.01), but the content of P decreased significantly (P<0.01); compared with the model group, hyperplasia of mammary gland was improved in all treatment groups, and serum E2, ER and PR positive expression of integral decreased significantly (P<0.05); the content of P in acupuncture group increased significantly (P<0.01), and the acupunture group and TCM low-dose group were the most obvious. Conclusion Rule Granules can effectively improve hyperplasia of mammary gland organization structure in rats caused by exogenous estrogen and progesterone, with TCM low-dose group and acupuncture group having the best efficacy. Its mechanism is related to reducing the contents of serum E2and ER, and PR positive expression in breast tissue, and increasing the content of serum P.
ABSTRACT
Objective:To study the action mechanism of Rukang oral liquid in rats with hyperplasia of mammary glands. Meth-ods:The rats were randomly divided into six groups:the normal group, the model group, tamoxifen group, low, medium and high dose of Rukang oral liquid groups. The model of hyperplasia of mammary glands was induced by estradiol benzoate injection and progester-one injection. After the administration, ELISA was used to detect the serum concentrations of ER, PR, P53 and Oct4, Western blot was used to detect the expression of ER, PR, P53 and Oct4 in breast tissue, a fluorescence quantitative PCR method was used to de-tect the expression of GnRH, GnRHR, and the expression of telomerase in breast tissue was detected by in situ hybridization. Results:Rukang oral liquid could reduce the expression of ER and PR in serum and breast tissue(P<0. 01), reduce the expression of P53, te-lomerase and Oct4(P<0. 01), and decrease the expression of GnRH in hypothalamus and pituitary, which could reverse the hyperpla-sia of mammary glands and played the role effectively(P<0. 01). Conclusion: Rukang oral Liquid can reduce the expression of ER and PR in serum and breast tissue, and decrease the expression of GnRH in hypothalamus and pituitary to play the role in the treatment of hyperplasia of mammary glands. Rukang oral Liquid may prevent breast hyperplasia from developing into breast cancer through re-ducing the expression of three related markers P53, telomerase and Oct4.
ABSTRACT
OBJECTIVE:To explore the effect of Shugan jianpi formula on the proliferation and apoptosis of the hyperplastic tissues of mammary gland of model rats.METHODS:The rats were injected with benzestrofol and progesterone to establish mammoplasia model,meanwhile which were treated intragastrically with Shugan jianpi formula for 30 days.The degree of mammoplasia,as well as the PCNA(proliferating cell nuclear antigen),the apoptotic related regulating factors such as Bcl-2 and Bax and the apoptotic index(AI)of the mammary gland were detected.RESULTS:Shugan jianpi formula inhibited the mammoplasia,down-regulated the expression levels of PCNA and Bcl-2,up-regulated the expression level of Bax and AI,inhibited the proliferation of the mammary tissues and promoted apoptosis.CONCLUSION:Shugan jianpi formula blocks or reverses the hyperplasia of mammary gland by regulating the cell proliferation/apoptosis of the hyperplastic tissues of mammary gland.
ABSTRACT
Objective To investigate the prevention and treatment mechanism of liver-smoothing and spleen-srengthening formula on hyperplasia of mammary gland,we study the effects of it on morphology,histology,serum sex hormone and its receptor of rats with hyperplasia of mammary gland.Methods Sixty female Wistar rats(SPF)were randomized into 5 groups:control group,disease model group,the tamoxifen group,small dose of medicine group and large-dose drug group.Estradiol Benzoate and Progesterone were used to establish animal model of breast hyperplasia.Rats in different groups were respectively maintained with Tamoxifen,small and large dose of liver-smoothing and spleen-srengthening formula.Then changes of morphology,histology,serum sex hormone and its receptor were obtained.Results Liver-smoothing and spleen-srengthening formula could reduce the number of breast lobular and the expansion degree of adenovirus catheter.It also narrowed the acinar cavity.In serum of rats with breast hyperplasia,Estradiol(E2)and PRL were declined ;Progesterone(P)and Testosterone(T)were increased;FSH and LH were partly increased.Positive expression degree of ER and PR were inhibited obviously.Conclusion Liver-smoothing and spleen-srengthening formula can reduce not only the content of estrogen receptors(ER)and progesterone receptors(PR)but also their sensitivity to estrogen of rats with breast hyperplasia by regulating the serum sex hormone.Our studies suggest that it can obviously inhibit hyperplasia of mammary gland.