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1.
Chinese Journal of Experimental Traditional Medical Formulae ; (24): 164-172, 2022.
Artículo en Chino | WPRIM | ID: wpr-940741

RESUMEN

ObjectiveTo predict the underlying mechanism of Bushen Huoxuetang in treating osteoporosis related to endocrine therapy in breast cancer by network pharmacology and to verify the results through in vitro cell model. MethodThe main effective components and targets of Bushen Huoxuetang were screened out through network pharmacology, and the targets of osteoporosis related to endocrine therapy in breast cancer were further obtained. The intersected targets were analyzed by Gene Ontology (GO) functional annotation and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment. Kaplan Meier plotter was used to analyze the survival of crucial targets. Finally, the inhibitory activity against cell proliferation was evaluated by in vitro methye thiazolye telrazlium(MTT) assay. The key targets and pathways were verified by Western blot, and the mRNA expression of the key targets was evaluated by real-time polymerase chain reaction(Real-time PCR). ResultA total of 716 active components and 249 key targets of Bushen Huoxuetang were obtained from network pharmacology. There were 135 common targets, among which protein kinase B(Akt)1 and hypoxia-inducible factor-1α (HIF-1α) were two key targets. Additionally, 531 biological processes, 62 cellular components, 162 molecular functions, and 145 signaling pathways including breast cancer and endocrine resistance were involved. The key targets were effectively enriched in phosphatidylinositol 3-kinases(PI3K)/Akt and HIF-1 signaling pathways. According to the MTT assay, the cell proliferation rate and cell motility of MCF-7 and T47D cells in the luminal A cell line were reduced by Bushen Huoxuetang treatment (22.5, 45, 90 g·L-1, and 45, 90, 180 g·L-1) for 48 h as compared with the blank group. As revealed by Western blot, MCF-7 cells were treated with Bushen Huoxuetang (0, 15, 60 g·L-1) for 48 h, and the relative expression of p-PI3K, PI3K, p-Akt, Akt, and HIF-1α was decreased in a dose-dependent manner as compared with the blank group (P<0.05, P<0.01). Real-time PCR was used to detect the mRNA expression of the key target HIF-1α. The results showed that the mRNA expression of HIF-1α in MCF-7 cells was decreased with the increase in the dose (P<0.01), and the change was in a concentration-dependent manner. ConclusionThe mechanism of Bushen Huoxuetang in the treatment of osteoporosis related to endocrine therapy in breast cancer may be related to the key targets including Akt1 and HIF-1α through the PI3K/Akt/HIF-1α signaling pathway.

2.
Chinese Journal of Experimental Traditional Medical Formulae ; (24): 220-228, 2019.
Artículo en Chino | WPRIM | ID: wpr-802444

RESUMEN

Diabetes mellitus (DM) is a metabolic disease characterized by hyperglycemia, with a high incidence and many complications. It has become an increasingly serious public health problem in the world, and has seriously affected the quality of life. Phosphatidylinositol-3-kinases (PI3K)/protein kinase B (Akt) signaling pathway is the main pathway of insulin signal transmission and the main signal channel for regulating blood glucose. The abnormal signal molecule of PI3K/Akt may cause abnormal signal transduction pathway, so as to impact the proliferation, apoptosis, metastasis and invasion of the corresponding tissues and organs, and lead to the occurrence of disease. Study of PI3K/Akt signal channel has a positive significance for investigating whether traditional Chinese medicine(TCM) has a definite and stable hypoglycemic effect. Currently, there are many TCM and Western medicines to treat diabetes, however, most drugs, especially Western medicines, have a relatively poor effect in controlling complications. To understand the progress of TCM in treatment of diabetes, in expectation of better studying the comprehensive therapeutic effect and mechanism of TCM on diabetes, and further developing the multi-target, multi-way and multi-channel advantages and features of TCM in the treatment of diabetes, this paper focuses on a systematic analysis on the progress of in vivo and in vitro studies on DM based on PI3K/Akt signaling channel in recent years, including the effect of the signaling channel on insulin secretion, the three main target organs of insulin (liver, skeletal muscle and fat), and its effect on the four main complications of diabetes (brain, kidney, heart, testis), and also provides certain ideas and guidance for the study of hypoglycemic mechanism of TCM monomer, TCM and compound medicine.

3.
Chinese Journal of Experimental Traditional Medical Formulae ; (24): 67-72, 2019.
Artículo en Chino | WPRIM | ID: wpr-802200

RESUMEN

Objective:To study the protective effect of tetramethylpyrazine (TMP) on PC12 cells induced by tert-butyl hydroperoxide (t-BHP) and the regulatory mechanism on signaling pathway of phosphatidylinositol-3-kinases (PI3K)/kinase B (Akt)/mammalian target of rapamycin(mTOR). Method:PC12 cells cultured in vitro were treated with t-BHP (200 μmol·L-1) for 6 h to establish a model of oxidative damage in PC12 cells. The experiment was divided into blank group, model group (200 μmol·L-1t-BHP), TMP group. PC12 cells were pretreated with TMP(25, 50, 100 μmol·L-1) for 12 h, and then treated with t-BHP for 6 h. The cell viability was detected by cell counting kit-8(CCK-8) method, and lactate dehydrogenase (LDH) leakage, malondialdehyde (MDA) content, superoxide dismutase (SOD) activity, reactive oxygen species (ROS) and glutathione peroxidase (GSH-Px) activity were detected by enzyme-linked immunosorbent assay (ELISA). Apoptosis was observed by annexin V-FITC/PI double staining. B-cell lymphoma-2 (Bcl-2), Bcl-2-associated X protein (Bax), total protein kinase B (Akt), and phosphorylated protein kinase B (p-Akt), mTOR and p-mTOR expressions were detected by Western blot. Result:The cell viability of PC12 cells treated with 200 μmol·L-1 t-BHP decreased to about 50%after 6 h. This condition was suitable for the establishment of oxidative damage model. Compared with the model group, TMP (25, 50, 100 μmol·L-1) pretreatment for 12 h significantly increased the survival rate of PC12 cells (PPPPPPP-1) pretreatment group increased significantly (PConclusion:Ligustrazine protects PC12 cell injury induced by t-BHP by activating PI3K/Akt/mTOR signaling pathway.

4.
Chinese Journal of Experimental Traditional Medical Formulae ; (24): 25-35, 2019.
Artículo en Chino | WPRIM | ID: wpr-801861

RESUMEN

Objective:Exploring the material basis and mechanism of Wuzi Yanzongwan in the treatment of male infertility based on network pharmacology. Method:Traditional Chinese medicine systems pharmacology database and analysis platform(TCMSP) was used to screen the active ingredients and targets in Wuzi Yanzongwan. GeneCards, OMIM and PharmGkb databases were used to screen the targets of male infertility. R language software was used to screen common targets of drugs and diseases, Pharmaceutical active ingredients-disease target interaction network was constructed by using Cytoscape software. The common target protein interaction network (PPI) was constructed by STRING platform, the gene ontology(GO) analysis of common target was analyzed by ClueGo plug-in, and Kyoto encyclopedia of genes and genomes(KEGG) pathway was enriched by R language software. Result:A total of 72 active ingredients were obtained from Wuzi Yanzongwan, and 35 possible targets for the treatment of male infertility were obtained. These targets are mainly involved in biological processes such as oxidation and antioxidant activity, and are mainly concentrated in phosphatidylinositol 3-kinase/protein kinase B(PI3K/Akt) and hypoxia inducible factor-1(HIF-1) signaling pathways. Conclusion:The network pharmacology confirmed the multi-component, multi-target and multi-pathway action characteristics of Wuzi Yanzongwan, predicted the possible mechanism of Wuzi Yanzongwan in the treatment of male infertility, and provided theoretical basis for further study of its active ingredients and mechanism.

5.
Chinese Journal of Experimental Traditional Medical Formulae ; (24): 101-106, 2019.
Artículo en Chino | WPRIM | ID: wpr-801701

RESUMEN

Objective: To research the effect of Wendantang on phosphatidylinositol-3-kinases(PI3K)/protein kinase B(Akt)/glycogen synthase kinase 3β(GSK3β) signaling pathway of hippocampus in rats with schizophrenia. Method: Sixty SD rats were randomly divided six groups:normal group (A), model group (B), Clozapine group (C), high-dosage group Wendantang (D), medium-dosage Wendantang group (E) and low-dosage Wendantang group (F), with 10 rats in each group. The rats of normal group and model group were given normal saline. The rats of Clozapine group were given 20 mg·kg-1 Clozapine. The rats of high-dosage, medium-dosage, low-dosage Wendantang groups were respectively given 40,20,10 g·kg-1 Wendantang, once a day, for 21 days.Two hours later after the last administration with Wendantang or saline,except for group A, groups B,C,D,E,F were intraperitoneally given MK-801 0.6 mg·kg-1 to establish the rat schizophrenia model. After modeling, the changes in stereotyped behavior of each group were observed and recorded. After three days,the rats were put to death,and the hippocampus tissue were tested. According to Sams Dodd and Hoffman's standard, the stereotyped behavior of rats was scored. The protein expressions of PI3K, Akt and GSK3β of hippocampus were determined by Western blot. The mRNA expressions of PI3K, Akt and GSK3β of hippocampus were tested by Real-time PCR. Result: Compared with A, the stereotyped behavior score of the B was significantly increased (Pβ protein and mRNA of hippocampus were significantly decreased (PPβ protein and PI3K, Akt and GSK3β mRNA of hippocampus (PPConclusion: Wendantang could regulate the PI3K/Akt/GSK3 signaling pathway by improving the stereotyped behavior and increasing the expressions of PI3K, Akt, GSK3β of hippocampus in rats with schizophrenia, so as to achieve the purpose of treating schizophrenia.

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