Effect and mechanism of Xihuang Pills on rats with precancerous lesions of breast / 中国中药杂志

The purpose of this study was to explore the effect and mechanism of Xihuang Pills on rats with precancerous lesions of the breast. Of 48 healthy female rats, 8 were randomly selected as blank group, and the other 40 were treated with 7,12-dimethylbenzanthracene(DMBA) combined with estrogen and progestin to establish a model of precancerous lesions of the breast. The successfully modeled rats were randomly divided into a model group, a tamoxifen group(1.8 mg·kg~(-1)·d~(-1)), a Xihuang Pills low-dose group(0.3 g·kg~(-1)·d~(-1)), a medium-dose group(0.6 g·kg~(-1)·d~(-1)) and a high-dose group(1.2 g·kg~(-1)·d~(-1)). After 30 days of admi-nistration, the histopathological changes of viscera and breast were observed by haematoxylin and eosin(HE) staining, and the visceral index was calculated. Enzyme linked immunosorbent assay(ELISA) was used to detect the contents of estradiol(E_2) and progesterone(P) in serum. The protein expressions of vascular endothelial growth factor(VEGF) and fibroblast growth factor 2(FGF2) were detected by immunohistochemistry. The protein expressions of VEGF, vascular endothelial growth factor receptor 2(VEGFR2), phosphorylated-vascular endothelial growth factor receptor 2(p-VEGFR2), B-cell lymphoma-2(Bcl-2), and Bcl-2 associated X protein(Bax) were detected by Western blot and the mRNA expressions of VEGF, FGF2, CXC-chemokine receptor 4(CXCR4), cysteine aspartic acid-specific protease(caspase-3), and stromal cell-derived factor 1(SDF-1) were detected by real-time polymerase chain reaction(RT-PCR). HE staining revealed that the model group had some liver and kidney damages and severe hyperplastic mammary tissue, while the Xihuang Pills high-dose group had mild hyperplasia. Compared with the model group, the Xihuang Pills groups had lo-wer ovarian coefficient(P<0.05 or P<0.01) and Xihuang Pills high-dose group had lower uterine coefficient(P<0.01). ELISA results showed that compared with the model group, expressions of E_2 and P in Xihuang Pills high-dose group were significantly decreased(P<0.05 or P<0.01). Immunohistochemistry, Western blot and RT-PCR indicated that compared with the conditions in the model group, the protein and mRNA expressions of VEGF and FGF2 in the Xihuang Pills groups were down-regulated(P<0.05 or P<0.01), and the protein expression of Bcl-2 was lowered(P<0.01); there was a decrease in the protein expressions of VEGFR2 and p-VEGFR2(P<0.01), a down-regulation in the mRNA expressions of CXCR4 and SDF-1(P<0.01), while an increase in the mRNA expression of caspase-3(P<0.01) in both Xihuang Pills medium-dose and high-dose groups; the protein expression of Bax in Xihuang Pills high-dose group was increased(P<0.01). The above results indicated that Xihuang Pills can effectively intervene in precance-rous lesions of the breast, and the mechanism may be related to the regulation of E_2 and P secretion as well as the inhibition of angiogenesis and chemokine receptor expression, thus controlling the occurrence of precancerous lesions of the breast in rats.

Mechanism of anti-hyperplasia of mammary glands of Xihuang Pills blood-entering component based on UPLC-Q-TOF-MS and network pharmacology / 中国中药杂志

In this study, based on network pharmacology and molecular docking method, the mechanism of anti-hyperplasia of mammary glands of Xihuang Pills blood-entering components was explored, and the efficacy and key targets of Xihuang Pills blood-entering components were experimentally verified by MCF-10A proliferation model of human mammary epithelial cells. In order to clarify the material basis and mechanism of Xihuang Pills in realizing anti-hyperplasia of mammary glands, the blood-entering components of Xihuang Pills were qualitatively analyzed by UPLC-Q-TOF-MS, and 22 blood-entering components were identified. By taking the blood-entering components as the research object, the network pharmacology prediction and molecular docking verification were carried out, and finally, three key targets were screened out, namely JAK1, SRC, and CDK1. In vitro experiments show that Xihuang Pills can inhibit the proliferation of MCF-10A cells, promote the apoptosis of MCF-10A cells, and reduce the expression of JAK1, SRC, and CDK1 targets in cells. To sum up, Xihuang Pills can promote the apoptosis of mammary epithelial cells by regulating the expression of JAK1, SRC, and CDK1 and then play an anti-hyperplasia role, which provides an experimental basis for clarifying the material basis of Xihuang Pills for anti-hyperplasia effect.

Urine metabolomics study of intervention of Xihuang Pills in rats with hyperplasia of mammary gland / 中国中药杂志

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.

Effects of Xihuang Pills on proliferation and apoptosis of prostate cancer LNCaP cells based on AR/m TOR signaling pathway / 中国中药杂志

Based on the androgen receptor(AR)/mammalian target of rapamycin(mTOR)signaling pathway, the effects of Xihuang Pills-medicated serum on the proliferation and apoptosis of prostate cancer LNCaP cells were investigated. The drug-containing serum of SD rats was prepared by intragastric administration of Xihuang Pills suspension. The effects of low-, medium-, and high-dose Xihuang Pills-containing serum on the in vitro proliferation of LNCaP cells were detected by cell counting kit-8(CCK-8). Flow cytometry was used to detect the apoptosis level of LNCaP cells after intervention with different concentrations of Xihuang Pills. Protein expression of cleaved cysteinyl aspartate-specific proteinase caspase-3(cleaved caspase-3), B-cell lymphoma-2(Bcl-2), and AR as well as the phosphorylation level of mTOR protein were detected by Western blot. The results showed that compared with the blank serum, the drug-medicated serum could blunt the activity of LNCaP cells. Low-, medium-, and high-dose Xihuang Pills-containing serum could significantly increase the cell apoptosis rate, increase the expression of cleaved caspase-3 protein, decrease the expression of Bcl-2 protein, reduce the expression of AR protein, and down-regulate the level of phosphorylated mTOR(p-mTOR). To study the effect of Xihuang Pills on the growth of LNCaP cells in vivo, different doses of Xihuang Pills were used to intervene in the subcutaneous graft model in nude mice inoculated with LNCaP cells. The expression levels of AR, mTOR, p-mTOR, Bcl-2, and cleaved caspase-3 were detected by Western blot. The results showed that the volumes of subcutaneous graft tumor in the low-dose, medium-dose, and high-dose Xihuang Pills groups significantly decreased compared with that in the model group. The weight of subcutaneous transplanted tumor in each group with drug intervention was significantly lower than that in the model group. Compared with the model group, the low-dose, medium-dose, and high-dose Xihuang Pills groups showed increased cleaved caspase-3 protein expression, decreased Bcl-2 and AR protein expression, and reduced p-mTOR protein expression. Further experiments showed that AR agonist R1881 could block the anti-proliferation and pro-apoptotic effects of Xihuang Pills. The mechanism of Xihuang Pills against prostate cancer is related to the inhibition of the AR/mTOR signaling pathway, inhibition of LNCaP cell proliferation, and induction of apoptosis in cancer cells.

Application of quantitative analysis of multi -components by single marker combined with fingerprint and chemical pattern recognition analysis in the quality evaluation of Xihuang pills / 中国药房

OBJECTIVE To evaluate the qualit y of Xihuang pills ，and to screen the differential markers affecting its quality . METHODS Using muskone as internal reference ，the content of α-pinene and other 4 components were determined by quantitative analysis of multi -components by single marker （QAMS），and compared with the results of external standard method . The fingerprints of 13 batches of Xihuang pills were established by gas chromatography （GC）method. Cluster analysis （CA）and orthogonal partial least squares discriminant analysis （OPLS-DA） were performed by SPSS 25.0 software and SIMCA 14.1 software. The variable importance projection （VIP）value greater than 1 was used as the standard to screen differential markers affecting the quality of the samples . RESULTS The contents of α-pinene，octyl acetate and β-elemene measured by QAMS were 0- 0.628 4，0.378 0-2.679 4 and 0.320 9-0.815 4 mg/g，respectively. The contents of α-pinene，octyl acetate ，β-elemene and musk ketone measured by external standard method were 0.001 5-0.627 1，0.378 0-2.594 7，0.329 2-0.837 0 and 0.385 7-0.806 0 mg/g， respectively. The relative error of the content determination results of the two methods was less than 4%. There were 26 common peaks in 13 batches of Xihuang pills ，and 3 common peaks ，such as octyl acetate ，β-elemene and musk ketone ，were identified ； their similarities were 0.912-0.946. 13 batches of samples could be divided into two categories （S1-S2，S6-S10，S13 were clustered into one category and S 3-S5，S11-S12 were clustered into one category ）. VIP values of peak 7，11，10，17 and 16 were all greater than 1. CONCLUSIONS The content of 4 components such as α-pinene in Xihuang pills combined with GC fingerprint and chemical pattern recognition analysis can be used to evaluate the quality of Xihuang pills . The components corresponding to 5 common peaks such as peak 7 may be differential markers affecting the quality of the samples .

Effect of Xihuang pills on apoptosis and expression of ER and PR in rat mammary epithelial cells / 中国药理学通报

Aim To investigate the effect of Xihuang pills on estradiol (E2) and progesterone (P)-induced apoptosis of rat mammary epithelial cells and the expression of estrogen and progesterone receptors, and to explore its anti-mammary hyperplasia mechanism. Methods Rat mammary epithelial cells were treated with different concentrations of Xihuangpills containing serumafter E2, P induced proliferation. The cell proliferation was detected by CCK-8 method (24, 48, 72 h). The apoptosis was detected by flow cytometry after AnnexinV-FITC/PI double staining. The average optical density of Bax and Bcl-2 was detected by immunohistochemistry; Estradiol receptor alpha (ER-α), estradiol receptor beta (ER-β), progesterone receptor (PR) protein expression were detected by Western blot. Results The serum containing Xihuang pillssignificantly inhibitedthe proliferation of rat mammary epithelial cells after E2and P treatment (P < 0. 05), and induced apoptosis, affecting the expression of apoptosis-related proteins Bcl-2 and Bax (P < 0. 05), and effectively inhibiting the expression of estrogen receptors ERα, ERβ. Conclusions Xihuang pills can induce the apoptosis of mammary epithelial cells by regulating related apoptotic proteins, and regulate the secretion of estrogen and progesterone in breast tissues, affecting the proliferation and rejuvenation of breast, which is of great significance for the treatment of breast hyperplasia.

Clinical observation of the effects of Xihuang pills combined with sodium cantharidinate and vitamin B6 injection in the treatment of tumor-induced fever / 国际中医中药杂志

Objective To study the effects of Xihuang pills combined with sodium cantharidinate and vitamin B6 injection in the treatment of tumor-induced fever. Methods A total of 100 patients with tumor-induced fever in our hospital from March 2015 to January 2017 were enrolled in this study. The subjects were divided into the control group (n=50) and the treatment group (n=50) randomly. The control group were treated with aspirin-D L-Lysine injection, and the treatment group were treated with Xihuang pills combined with sodium cantharidinate and vitamin B6 injection. The two groups were treated for 2 periods. The clinical effects of the two groups after treatment were compared. The life quality of the two groups after treatment were compared. The serum TNF-α and IL-1β of the two groups before and after treatment were compared. The adverse reaction rates of the two groups during treatment were compared. Results The total efficacy rate of defervescence of the treatment group was 88.0％(44/50) significantly higher than 56.0％(28/50) of the control group (χ2=12.698, P=0.000). After treatment, the daily life score (1.10 ± 0.18 vs. 2.47 ± 0.21, t=35.025), social communication score (1.21 ± 0.13 vs. 2.53 ± 0.25, t=33.124), mental state score (1.08 ± 0.15 vs. 2.75 ± 0.21,t=45.758), appetite score (1.13 ± 0.16 vs. 2.56 ± 0.19, t=40.708), sleep score (1.22 ± 0.17 vs. 2.71 ± 0.20, t=40.139) of the treatment group were significantly lower than those of the control group (P<0.05). After treatment, the serum TNF-α (1.98 ± 0.07 μg/L vs. 2.86 ± 0.13 μg/L, t=42.144), IL-1β (9.20 ± 1.89 μg/L vs. 13.51 ± 2.36 μg/L, t=10.080) of the treatment group were significantly lower than those of the control group (P<0.05). There was no significantly differences of the adverse reaction rates of the two groups during treatment (χ2=0.211, P=0.646). Conclusions The Xihuang pills combined with sodium cantharidinate and vitamin B6 injection for patients with tumor-induced fever has a good efficacy and low incidence of adverse reactions, can improve the quality of life and reduce the serum levels of TNF-α and IL-1β.

Study on fingerprint analysis of Xihuang Pills by HPLC-ELSD / 中草药

Objective: To establish the quality evaluation of fingerprints of Xihuang Pills by HPLC-ELSD. Methods: The chromatography conditions were defined as waters C18 column (250 mm × 4.6 mm, 5 μm); Mobile phase was methanol-0.5% acetic acid, gradient elution; temperature of column was set at 30℃. The ELSD conditions were as follows: the temperature of drift tube was 45℃, the gas speed was 1.5 L/min. Ten batches of Xihuang Pills samples were analyzed for similarity analysis (SA), hierarchical clustering analysis (HCA) and principle component analysis (PCA). Results: The chromatographic fingerprint was completed with 25 recognizable peaks, and the samples with great differences and the compounds with greater impact on the quality were obtained through HCA and PCA. Conclusion: HPLC fingerprint combining with pattern recognition could reflect the intrinsic quality to provide a scientific basis for the quality control of Xihuang Pills.

On treatment of angina pectoris in coronary heart disease with Xihuang Pills / 中华中医药杂志

0.05).Conclusion:Treatment of angina pectoris with syndrome of accumulation phlegm-heat in the interior with cariac blood stasis by Xihuang Pills was safe and effective.

Integrated Xihuang Pills and chemotherapy in treating 30 patients with breast cancer / 中华中医药杂志

Objective:To observe the clinical efficacy of Xihuang Pills combining with chemotherapy in treating breast cancer and the effects on CD+4,CD+8.Methods:Selecting 60 patients with breast cancer,the control group of 30 was treated with chemotherapy,the treated group of 30 was treated with Xihuang Pills combining with chemotherapy for two chemotherapy cycles,before and after treatment the CD+4,CD+8 levels were detected.Results:The traditional Chinese medicine syndrome and immune function in treated group was superior to those in control group,the difference between those two groups was statistically significant(P

Clinical study of Xihuang Pills in treating cyclomastopathy / 中华中医药杂志

Objective: To explore the clinical efficacy of Xihuang Pills in treating cyclomastopathy. Methods: Based on inclusion criteria and exclusion criteria, 56 patients with cyclomastopathy were selected, 30 ones in treatment group and take the Xihuang Pills, the other 26 ones were in control group and took Rupixiao Tablets for 3 courses of treatment. Results: The totaleffective rate was 83.33% in the treatment group and that in the control group was 69.22%. The difference between the two groups was statistically significant (P