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A Novel Based-Network Strategy to Identify Phytochemicals from Radix Salviae Miltiorrhizae (Danshen) for Treating Alzheimer's Disease.

ABSTRACT: Alzheimer's disease (AD) is a common age-related neurodegenerative disease that strikes millions worldwide. Herein, we demonstrate a new approach based on network target to identify anti-AD compounds from Danshen. Network pharmacology and molecular docking were employed to establish the DS-AD network, which mainly involved apoptosis of neuron cells. Then network scoring was confirmed via Connectivity Map analysis. M308 (Danshenxinkun D) was an anti-AD candidate with a high score (p < 0.01). Furthermore, we conducted ex vivo experiments with H₂O₂-treated PC12 cells to verify the neuroprotective effect of Salvia miltiorrhiza-containing plasma (SMP), and UPLC-Q-TOF/MS and RT-qPCR were performed to demonstrate the anti-AD activity of M308 from SMP. Results revealed that SMP could enhance cell viability and level of acetylcholine. AO/EB staining and Mitochondrial membrane potential (MMP) analysis showed that SMP significantly suppressed apoptosis, which may be due to anti-oxidative stress activity. Moreover, the effects of M308 and SMP on expressions of PSEN1, DRD2, and APP mRNA were consistent, and M308 can significantly reverse the expression of PSEN1 and DRD2 mRNA in H₂O₂-treated PC12 cells. The strategy based on the network could be employed to identify anti-AD compounds from Chinese herbs. Notably, M308 stands out as a promising anti-AD candidate for development.

SUBMITTER: Li B

PROVIDER: S-EPMC9317794 | biostudies-literature |

REPOSITORIES: biostudies-literature

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A network pharmacology-based study on the anti-hepatoma effect of Radix Salviae Miltiorrhizae.

Project description:BackgroundRadix Salviae Miltiorrhizae (RSM), a well-known traditional Chinese medicine, has been shown to inhibit tumorigenesis in various human cancers. However, the anticancer effects of RSM on human hepatocellular carcinoma (HCC) and the underlying mechanisms of action remain to be fully elucidated.MethodsIn this study, we aimed to elucidate the underlying molecular mechanisms of RSM in the treatment of HCC using a network pharmacology approach. In vivo and in vitro experiments were also performed to validate the therapeutic effects of RSM on HCC.ResultsIn total, 62 active compounds from RSM and 72 HCC-related targets were identified through network pharmacological analysis. RSM was found to play a critical role in HCC via multiple targets and pathways, especially the EGFR and PI3K/AKT signaling pathways. In addition, RSM was found to suppress HCC cell proliferation, and impair cancer cell migration and invasion in vitro. Flow cytometry analysis revealed that RSM induced cell cycle G2/M arrest and apoptosis, and western blot analysis showed that RSM up-regulated the expression of BAX and down-regulated the expression of Bcl-2 in MHCC97-H and HepG2 cells. Furthermore, RSM administration down-regulated the expression of EGFR, PI3K, and p-AKT proteins, whereas the total AKT level was not altered. Finally, the results of our in vivo experiments confirmed the therapeutic effects of RSM on HCC in nude mice.ConclusionsWe provide an integrative network pharmacology approach, in combination with in vitro and in vivo experiments, to illustrate the underlying therapeutic mechanisms of RSM action on HCC.

| S-EPMC6685170 | biostudies-literature

Genotoxicity Evaluation of an Ethanol Extract Mixture of Astragali Radix and Salviae miltiorrhizae Radix.

Project description:Myelophil, a combination of Astragali Radix and Salviae Radix, is one of the most commonly used remedies for disorders of Qi and blood in traditional Chinese medicine. Based on the clinical applications of these plants, in particular to pregnant woman, this study aimed to evaluate the genotoxic potential of an ethanol extract mixture of the above two herbs, called Myelophil. Following the Organization for Economic Cooperation and Development (OECD) Guideline methods, a genotoxicity test was conducted using a bacterial reverse mutation test with Salmonella typhimurium (TA98, TA100, TA1535, and TA1537) and Escherichia coli (WP2?vrA), an in vitro mammalian chromosome aberration test using a Chinese hamster ovary cell line (CHO-K1), and an in vivo mammalian erythrocyte micronucleus test using ICR mouse bone marrow. In the Ames test, for both types of mutations (base substitution and frameshift) under conditions with/without an S9 mix up to 5,000??g/plate, Myelophil did not increase the number of revertant colonies of all S. typhimurium strains as well as E. coli strain. For both short (6?h) and long tests with/without S9 mix, the chromosome aberration test did not show any significant increase in the number of structural or numerical chromosome aberrations by Myelophil. In addition, no significant change in the number of micronucleated polychromatic erythrocytes or polychromatic erythrocytes was observed in the bone marrow of an ICR mouse administered Myelophil orally at 2,000?mg/kg/day for 2 days, respectively. These results are the first to provide experimental evidence that Myelophil, an ethanol extract mixture of Astragali Radix and Salviae Radix, has no risk of genotoxicity.

| S-EPMC6198562 | biostudies-literature

Effect of methanol extract of Salviae miltiorrhizae Radix in high-fat diet-induced hyperlipidemic mice.

Project description:BACKGROUND:The dried root of Salvia miltiorrhiza, Salviae miltiorrhizae Radix (SR), is one of the most popular medicinal herbs in Asian countries such as China and Korea. In Asian traditional medicine, SR is considered to have a bitter flavor, be slightly cold in nature, and exert therapeutic actions in the heart and liver meridians. Thus, SR has been used to control symptoms related to cardiovascular diseases. Hyperlipidemia is recognized as the main cause of cerebrovascular and heart diseases; consequently, therapeutic strategies for hyperlipidemia have been widely studied. In this study, the effects and molecular targets of methanol extract of SR (SRme) in hyperlipidemic mice were investigated. METHODS:High-fat diet was fed to mice to induce hyperlipidemia, and measurement of blood cholesterol and triglycerides were conducted to evaluate the effect of SRme on hyperlipidemic mice, and gene expression in mice liver was analyzed to identify key molecules which could be potential targets for developing anti-hyperlipidemic herbal medicines. RESULTS:There was no significant effect on the body weight gain of hyperlipidemic mice, but the triglyceride content in blood was significantly reduced by the administration of SRme to hyperlipidemic mice. Proteins such as minichromosome maintenance (Mcm) family which play a key role in DNA replication were identified as molecular targets in the amelioration of hyperlipidemia. CONCLUSIONS:SRme ameliorated hyperlipidemia in high-fat diet fed mice by inhibiting increase of blood serum level of triglycerides. And several proteins such as Mcm proteins were deduced to be molecular targets in treating hyperlipidemia.

| S-EPMC5640945 | biostudies-literature

A Biomedical Investigation of the Hepatoprotective Effect of Radix salviae miltiorrhizae and Network Pharmacology-Based Prediction of the Active Compounds and Molecular Targets.

Project description:Radix salviae miltiorrhizae (Danshen in Chinese), a classic traditional Chinese medicine (TCM) herb, has been used for centuries to treat liver diseases. In this study, the preventive and curative potential of Danshen aqueous extract on acute/chronic alcoholic liver disease (ALD) and non-alcoholic fatty liver disease (NAFLD) was studied. The in vivo results indicated that Danshen could alleviate hepatic inflammation, fatty degeneration, and haptic fibrogenesis in ALD and NAFLD models. In the aspect of mechanism of action, the significant reduction in MDA levels in both ALD and NAFLD models implies the decreased levels of oxidative stress by Danshen. However, Danshen treatment could not activate the internal enzymatic antioxidant system in ALD and NAFLD models. To further explore the hepatoprotective mechanism of Danshen, an in silico-based network pharmacology approach was employed in the present study. The pharmacological network analysis result revealed that six potential active ingredients such as tanshinone iia, salvianolic acid b, and Danshensu may contribute to the hepatoprotective effects of Danshen on ALD and NAFLD. The action mechanism may relate with regulating the intracellular molecular targets such as PPAR?, CYP1A2, and MMP2 for regulation of lipid metabolism, antioxidant and anti-fibrogenesis by these potential active ingredients. Our studies suggest that the combination of network pharmacology strategy with in vivo experimental study may provide a forceful tool for exploring the mechanism of action of traditional Chinese medicine (TCM) herb and developing novel bioactive ingredients.

| S-EPMC5372635 | biostudies-literature

Deep Eutectic Solvent-Based Microwave-Assisted Method for Extraction of Hydrophilic and Hydrophobic Components from Radix Salviae miltiorrhizae.

Project description:Deep eutectic solvents (DESs) have attracted significant attention as a promising green media. In this work, twenty-five kinds of benign choline chloride-based DESs with microwave-assisted methods were applied to quickly extract active components from Radix Salviae miltiorrhizae. The extraction factors, including temperature, time, power of microwave, and solid/liquid ratio, were investigated systematically by response surface methodology. The hydrophilic and hydrophobic ingredients were extracted simultaneously under the optimized conditions: 20 vol% of water in choline chloride/1,2-propanediol (1:1, molar ratio) as solvent, microwave power of 800 W, temperature at 70 °C, time at 11.11 min, and solid/liquid ratio of 0.007 g·mL-1. The extraction yield was comparable to, or even better than, conventional methods with organic solvents. The microstructure alteration of samples before and after extraction was also investigated. The method validation was tested as the linearity of analytes (r² > 0.9997 over two orders of magnitude), precision (intra-day relative standard deviation (RSD) < 2.49 and inter-day RSD < 2.96), and accuracy (recoveries ranging from 95.04% to 99.93%). The proposed DESs combined with the microwave-assisted method provided a prominent advantage for fast and efficient extraction of active components, and DESs could be extended as solvents to extract and analyze complex environmental and pharmaceutical samples.

| S-EPMC6274102 | biostudies-literature

Acute and repeated toxicological study of Myelophil, an ethanol extract of a mixture of Astragali Radix and Salviae Miltiorrhizae Radix, in beagle dogs.

Project description:BackgroundTo evaluate the pharmaceutical safety of Myelophil, an ethanol extract of a mixture of Astragali Radix and Salviae Miltiorrhizae Radix, using both acute and repeated toxicological studies.MethodsA total of 40 beagle dogs (20 each male and female) were fed doses up to 5,000 mg/kg for the acute study and up to 1,250 mg/kg for the 13-week repeated dose toxicological study. Adverse effects were examined intensively by comparing the differences between normal and drug-administered groups using clinical signs, autopsies, histopathological findings, hematology, urinalysis, and biochemical analysis.ResultsNo mortality or drug-related clinical signs were observed in the Myelophil-treated groups, except for vomiting due to an excessive dose (5,000 mg/kg). Likewise, in the repeated toxicity test, compound-colored stools in the Myelophil-treated groups and soft stools in all groups, including the control, were observed. No drug-related abnormalities were found in the histopathology, hematology, urinalysis, and biochemical analyses for any doses of Myelophil.ConclusionThese results support the safety of Myelophil with a no observed adverse effect level (NOAEL) of 1250 mg/kg in beagle dogs, which corresponds to a human equivalent dose (HED) of 694 g/kg.

| S-EPMC6615141 | biostudies-literature

Synergistic Effects of Danshen (Salvia Miltiorrhizae Radix et Rhizoma) and Sanqi (Notoginseng Radix et Rhizoma) Combination in Angiogenesis Behavior in EAhy 926 Cells.

Project description:Background: This study investigated the combination effects of the Danshen and Sanqi herb pair on angiogenesis in vitro. Methods: Nine combination ratios of Danshen-Sanqi extracts (DS-SQ) were screened for their angiogenic effects in the human vascular endothelial EAhy 926 cell line via cell proliferation, cell migration and tube formation activities against the damage to the cells exerted by DL-homocysteine (Hcy) and adenosine (Ado). The type of interaction (synergistic, antagonistic, additive) between Danshen and Sanqi was analyzed using combination index (CI) and isobologram models. The angiogenic activities of key bioactive compounds from Danshen and Sanqi were tested in the same models. Results: DS-SQ ratios of 2:8 and 3:7 (50-300 µg/mL) potentiated angiogenic synergistic effects (CI < 1) in all three assays. The observed wound healing effects of DS-SQ 2:8 was significantly attenuated by phosphatidylinositol-3 kinases (PI3K), mitogen-activated protein kinase (MEK) and extracellular signal-regulated kinases (ERK) inhibitors which inferred the potential mechanistic pathways. Out of all the tested compounds, Notoginsenoside R1 from Sanqi exhibited the most potent bioactivity in cell proliferation assay. Conclusions: This study provides scientific evidence to support the traditional use of the Danshen-Sanqi combination for vascular disease, in particular through their synergistic interactions on previously unexamined angiogenic pathways.

| S-EPMC5750609 | biostudies-literature

Network Pharmacology and Molecular Docking Analysis on Molecular Targets and Mechanisms of "Chuanxiong Rhizoma: Radix Salviae miltiorrhizae" Herb Couples in the Treatment of Preeclampsia.

Project description:ObjectiveThe aim of the study is to explore the molecular mechanism of activating blood circulation and dispersing stasis herbs in the treatment of pre-eclampsia with Chuanxiong Rhizoma-Radix Salvia miltiorrhiza.MethodsThe chemical composition and targets of Chuanxiong Rhizoma-Radix Salvia miltiorrhiza were retrieved from the TCMSP database, and a PPI network was constructed for common genes. Subsequently, a graph of the "active component-target-action pathway" was plotted by Cytoscape 3.7.2 and a KEGG pathway enrichment was performed using the R language cluster profiler package. Molecular docking was conducted between the top five PPI targets of Chuanxiong Rhizoma-Radix Salvia miltiorrhiza.ResultsAccording to network pharmacology, there were 32 target genes, 60 active components, and 59 pathways in Chuanxiong Rhizoma-Radix Salvia miltiorrhiza, and its most evident effects were exerted on G-protein-coupled amine receptors and the neuroactive ligand-receptor interaction signaling pathway. Molecular docking indicated that the target protein had a good binding ability with the drugs.ConclusionChuanxiong Rhizoma-Radix Salvia miltiorrhiza have therapeutic effects in pre-eclampsia, as confirmed by the results of molecular biology analysis. Thus, the Chuanxiong Rhizoma-Radix Salvia miltiorrhiza regimen provides a basis for the treatment of pre-eclampsia using traditional Chinese medicine.

| S-EPMC9473876 | biostudies-literature

Synergistic study of a Danshen (Salvia Miltiorrhizae Radix et Rhizoma) and Sanqi (Notoginseng Radix et Rhizoma) combination on cell survival in EA.hy926 cells.

Project description:BACKGROUND:This study investigated the protective effects of the Danshen (DS) and Sanqi (SQ) herb pair on cell survival in the human cardiovascular endothelial (EA.hy926) cell line exposed to injury. METHODS:Nine combination ratios of Danshen-Sanqi extracts (DS-SQ) were screened for their protective effects in the EA.hy926 cell line against two different cellular impairments induced by DL-homocysteine (Hcy) - adenosine (Ado) - tumour necrosis factors (TNF) and oxidative stress (H2O2), respectively. The type of interaction (synergistic, antagonistic, additive) between DS and SQ was analysed using a combination index (CI) model. The effects of key bioactive compounds from DS and SQ were tested using the same models. The compound from each herb that demonstrated the most potent activity in cell viability was combined to evaluate their synergistic/antagonistic interaction using CI. RESULTS:DS-SQ ratios of 6:4 (50-300 μg/mL) produced synergistic effects (CI < 1) in restoring cell viability, reducing lactate dehydrogenase (LDH) leakage and caspase-3 expressions against Hcy-Ado-TNF. Additionally, DS-SQ 6:4 (50-150 μg/mL) was found to synergistically protect endothelial cells from impaired cellular injury induced by oxidative damage (H2O2) by restoring reduced cell viability and inhibiting excessive expression of reactive oxygen species (ROS). In particular, the combination of salvianolic acid A (SA) and ginsenoside Rb1 (Rb1) at 4:6 (1-150 μM) showed synergistic effects in preventing cytotoxic effects caused by Hcy-Ado-TNF (CI < 1). This simplified combination also demonstrated synergistic effects on H2O2-induced oxidative damage on EA.hy926 cells. CONCLUSIONS:This study provides scientific evidence to support the traditional use of the DS-SQ combination on protecting endothelial cells through their synergistic interactions.

| S-EPMC6385400 | biostudies-other

Rapid Determination of 30 Polyphenols in Tongmai Formula, a Combination of Puerariae Lobatae Radix, Salviae Miltiorrhizae Radix et Rhizoma, and Chuanxiong Rhizoma, via Liquid Chromatography-Tandem Mass Spectrometry.

Project description:Tongmai formula (TMF) is a herbal preparation composed of three traditional Chinese medicinal materials: Puerariae Lobatae Radix (Gegen), Salviae Miltiorrhizae Radix et Rhizoma (Danshen) and Chuanxiong Rhizoma (Chuanxiong). It has been used to treat cardiovascular diseases for decades. To develop a reliable and convenient analytical method for a comprehensive determination of polyphenols in TMF and the ascertainment of their chemical correlations with its herbal components, a method combining high-performance liquid chromatography with electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS) was developed and validated for rapid determination of 30 polyphenols in TMF and its three herbal components. The chromatographic separation was carried out on a Chromolith Fastgradient RP-18 endcapped 50-2 column using an optimized gradient elution. Statistical analysis of obtained data demonstrated that the method had a desirable linearity, precision, and accuracy, as well as excellent sensitivity. The obtained results indicated that, among the 30 polyphenols in TMF, 22 originated from Gegen, 6 originated from Danshen, and 2 originated from Chuanxiong. The major polyphenols in TMF have been identified as puerarin, mirificin, salvianolic acid B, salvianic acid A, 3'-hydroxypuerarin, 3'-methoxypuerarin, and salvianolic acid A, with a combined contribution of 19.2% of the preparation. The development and validation of this method will greatly facilitate future pharmacological studies of TMF and its herbal components, as well as polyphenols in cardiovascular therapies.

| S-EPMC6154678 | biostudies-literature

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