ABSTRACT
Ischemic preconditioning (IPC) is a potential intervention known to protect the heart against ischemia/reperfusion injury, but its role in the no-reflow phenomenon that follows reperfusion is unclear. Dihydrotanshinone I (DT) is a natural compound and this study illustrates its role in cardiac ischemic injury from the aspect of IPC. Pretreatment with DT induced modest ROS production and protected cardiomyocytes against oxygen and glucose deprivation (OGD), but the protection was prevented by a ROS scavenger. In addition, DT administration protected the heart against isoprenaline challenge. Mechanistically, PKM2 reacted to transient ROS via oxidization at Cys423/Cys424, leading to glutathionylation and nuclear translocation in dimer form. In the nucleus, PKM2 served as a co-factor to promote HIF-1α-dependent gene induction, contributing to adaptive responses. In mice subjected to permanent coronary ligation, cardiac-specific knockdown of Pkm2 blocked DT-mediated preconditioning protection, which was rescued by overexpression of wild-type Pkm2, rather than Cys423/424-mutated Pkm2. In conclusion, PKM2 is sensitive to oxidation, and subsequent glutathionylation promotes its nuclear translocation. Although IPC has been viewed as a protective means against reperfusion injury, our study reveals its potential role in protection of the heart from no-reflow ischemia.
ABSTRACT
Objective To evaluate therapeutic effects of dihydrotanshinone Ⅰ on hepatic fibrosis based on liver metabolomics method. Methods 28 rats were randomly divided into four groups including control group, hepatic fibrosis model group and dihydrotanshinone Ⅰ low dose group and dihydrotanshinone Ⅰ high dose group. The dihydrotanshinone Ⅰ treated groups received dihydrotanshinone Ⅰ for 28 days. The rat liver samples were collected and analyzed by liquid chromatography-mass spectrometer (LC-MS). The OPLS-DA pattern recognition analysis of metabolomics differences among the groups and therapeutic effects of dihydrotanshinone Ⅰ on hepatic fibrosis were evaluated. Results 38 metabolites were identified through liver metabolomics analysis. The possible mechanism of hepatic fibrosis was mainly involved glutathione metabolism, melatonin metabolism, amino acid metabolism, lipid metabolism and TCA cycle. The hepatic fibrosis induced by TAA was reversed by dihydrotanshinone Ⅰ. Conclusion Dihydrotanshinone Ⅰ provided satisfactory therapeutical effects on hepatic fibrosis through partially regulating the perturbed glutathione metabolism, melatonin metabolism, amino acid metabolism, lipid metabolism, TCA cycle.
ABSTRACT
@#Disrupted redox status primarily contributes to myocardial ischemia/reperfusion injury (MIRI). NRF2, the endogenous antioxidant regulator, might provide therapeutic benefits. Dihydrotanshinone-I (DT) is an active component in Salvia miltiorrhiza with NRF2 induction potency. This study seeks to validate functional links between NRF2 and cardioprotection of DT and to investigate the molecular mechanism particularly emphasizing on NRF2 cytoplasmic/nuclear translocation. DT potently induced NRF2 nuclear accumulation, ameliorating post-reperfusion injuries via redox alterations. Abrogated cardioprotection in NRF2-deficient mice and cardiomyocytes strongly supports NRF2-dependent cardioprotection of DT. Mechanistically, DT phosphorylated NRF2 at Ser40, rendering its nuclear-import by dissociating from KEAP1 and inhibiting degradation. Importantly, we identified PKC-δ-(Thr505) phosphorylation as primary upstream event triggering NRF2-(Ser40) phosphorylation. Knockdown of PKC-δ dramatically retained NRF2 in cytoplasm, convincing its pivotal role in mediating NRF2 nuclear-import. NRF2 activity was further enhanced by activated PKB/GSK-3β signaling via nuclear-export signal blockage independent of PKC-δ activation. By demonstrating independent modulation of PKC-δ and PKB/GSK-3β/Fyn signaling, we highlight the ability of DT to exploit both nuclear import and export regulation of NRF2 in treating reperfusion injury harboring redox homeostasis alterations. Coactivation of PKC and PKB phenocopied cardioprotection of DT in vitro and in vivo, further supporting the potential applicability of this rationale. Graphical abstract
ABSTRACT
Objective: To evaluate the quality of Zidan Huoxue Tablets (ZHT) based on HPLC fingerprint and multi-component content determination combined with chemical pattern recognition method. Methods: ACE Neptune-C18 column (250 mm × 4.6 mm, 5 μm) was used with acetonitrile-0.1% phosphoric acid aqueous solution as mobile phase at a flow rate of 1.0 mL/min. The detection wavelength was 275 nm and the column temperature was 25 ℃. A total of 18 batches of ZHT was analyzed, and the quality of ZHT was evaluated by the similarity evaluation system of traditional Chinese medicine chromatographic fingerprints combined with cluster analysis and principal component analysis. Results: The fingerprint of ZHT was established. Twenty-six common peaks were identified and nine of them were identified, including 2-sodium danshensu, 8-rosmarinic acid, 9-lithospermic acid, 10-salvianolic acid B, 12-salvianolic acid A, 21-dihydrotanshinone I, 22-cryptotanshinone, 23-tanshinone I, and 25-tanshinone IIA. The similarity of fingerprints of 18 batches of ZHT was between 0.93 and 1.00. The results of cluster analysis and principal component analysis were basically consistent with similarity results. After validating the multiple component quantitative analysis condition through methodology, the average recoveries were between 98.27% and 103.42%, and the RSD were in the range of 0.86%-2.53%. The content of sodium danshensu, rosmarinic acid, lithospermic acid, salvianolic acid B, dihydrotanshinone I, cryptotanshinone, tanshinone I, tanshinone IIA in 18 batches of ZHT were in the range of 0.149%-0.218%, 0.179%-0.225%, 0.222%-0.286%, 3.570%-6.399%, 0.048%-0.136%, 0.122%-0.309%, 0.061%-0.215%, 0.093%-0.413%, respectively. Conclusion: There is a certain quality difference between different batches of ZHT. Through the combination of fingerprinting, cluster analysis and principal component analysis, the quality of ZHT can be comprehensively evaluated. The establishment of this method can provide reference for the quality control and evaluation of ZHT.
ABSTRACT
Objective: To comprehensively compare and evaluate the composition of Salvia miltiorrhiza phloem and xylem samples based on fingerprint analysis and quantitative analysis of multi-components. Methods: In the present study, an accurate and reliable fingerprint approach was developed using high performance liquid chromatography for chemical comparison ofphloem and xylem samples of S. miltiorrhiza. Furthermore, eight bioactive compounds including four salvianolic acids and four tanshinones in S. miltiorrhiza phloem and xylem samples were simultaneously quantified. Moreover, chemometrics methods were performed to compare and discriminate the phloem and xylem based on the quantitative data. Results: The specific fingerprints of phloem and xylem of S. miltiorrhiza were obtained, and a total of 10 common peaks were marked. The quantitative and chemometrics analysis results indicated the content of chemical components in phloem and xylem samples of S. miltiorrhiza were notably different. Obviously, the content of tanshinones were notably different between phloem and xylem samples. The content of tanshinones were significant higher in phloem compared with xylem in S. miltiorrhiza. Conclusion: The fingerprint analysis and quantitative analysis of multi-components could be a well-acceptable strategy for chemical comparison of S. miltiorrhiza phloem and xylem.
ABSTRACT
BACKGROUND: 15,16-dihydrotanshinone I (DHTS) is a natural abietane diterpenoid that is mainly found in the roots of Salvia miltiorrhiza Bunge (Labiatae). DHTS exhibits a potential anti-proliferative effect in various human cancer cells. However, the mechanisms of action of DHTS as an anti-cancer agent have not been fully elucidated. Therefore, the present study investigated the anti-cancer effect of DHTS in terms of cell cycle regulation and the regulation of the AMP-activated protein kinase (AMPK)/Akt/mTOR signaling pathway in SK-HEP-1 human hepatocellular carcinoma cells. METHODS: The anti-proliferative effects of DHTS were evaluated by the sulforhodamine B assay in SK-HEP-1 cells. Cell cycle distribution was analyzed by flow cytometry. The elucidation of mechanisms of action such as the AMPK/AKT/mTOR and mitogen-activated protein kinase (MAPK) pathway was assessed by Western blot analysis. RESULTS: DHTS showed a significant anti-proliferative activity against SK-HEP-1 cells. DHTS induced cell cycle arrest in the G0/G1 phase, which was mediated by downregulation of cyclin D1, cyclin A, cyclin E, CDK4, CDK2, c-Myc and p-Rb expression and with increased expression of the CDK inhibitor p21. DHTS also activated the AMPK signaling. In addition, DHTS downregulated the Akt/mTOR and MAPK signaling pathways. CONCLUSIONS: Our results suggest that the anti-proliferative activity of DHTS might be associated with the induction of G0/G1 phase cell cycle arrest and regulation of AMPK/Akt/mTOR and MAPK signaling pathways in SK-HEP-1 cells.
Subject(s)
Humans , AMP-Activated Protein Kinases , Blotting, Western , Carcinoma, Hepatocellular , Cell Cycle Checkpoints , Cell Cycle , Cyclin A , Cyclin D1 , Cyclin E , Cyclins , Down-Regulation , Flow Cytometry , Protein Kinases , Salvia miltiorrhizaABSTRACT
Objective: To establish an HPLC method for fingerprint analysis of Guanxin Shutong Capsule (GSC) for its quality control. Methods: The chromatographic behaviors were obtained by an Agilent TC-C18 column (250 mm × 4.6 mm, 5 μm) with gradient elution using 0.05% phosphoric acid and acetonitrile as the mobile phase. The column temperature was set at 30 ℃ and the flow rate was 1.0 mL/min. The detection wavelength was set at 280 nm. The common mode of HPLC fingerprint for 10 batches of GSC was established with Similarity Evaluation System for Chromatographic Fingerprint of Traditional Chinese Medicine (2009 edition) and the common peaks were identified by reference compounds. Results: Fingerprints of 10 batches of GSC were established and the similarities to the common mode were above 0.95. Totally 45 common peaks were found, and 34 belonged to herbs. Five mutual peaks were from Choerospondias axillaris, 23 mutual peaks were from Salvia miltiorrhiza, and seven mutual peaks were from Syzigium aromaticum. Based on the retention time of reference substances, 13 constituents including gallic acid (peak 1), danshensu (peak 3), protocatechuic acid (peak 5), procatechuic aldehyde (peak 6), ellagic acid (peak 10), rosmarinic acid (peak 16), salvianolic acid A (peak 19), salvianolic acid B (peak 23), eugenol (peak 24), dihydrotanshinone I (peak 30), cryptotanshinone (peak 34), tanshinone I (peak 35), and tanshinone IIA (peak 39) were identified. Conclusion: This method is specific and reliable, which will be conducive to the quality control of GSC.
ABSTRACT
Objective: To establish a quantitative analysis method of multi-components by single marker (QAMS) for the simultaneous determination of four tanshinones [tanshinone IIA (TIIA), dihydrotanshinone I (DTI), cryptotanshinone (CT), and tanshinone I (TI)], and to explore its accuracy and feasibility on quality evaluation of traditional chinese medicine preparations about three kinds of related preparations of Salvia miltiorrhiza. Methods: An HPLC method was used to calculate relative correction factors (fi/s) between other three components with TIIA. The component positioning methods were compared including retention time difference value, relative retention value, and linear regression methods. Meanwhile, the contents of tanshinones in different preparations of S. miltiorrhiza were determinated by the external standard method (ESM), including Danshen Tablet, Compound Danshen Tablet, and Guanxin Danshen Capsule. Measured content value was compared with calculated content value to evaluate the accuracy of QAMS method. Results: Within a certain range, the fi/s of DTI, TI, and CT to TIIA were 0.734, 0.916, and 0.774, respectively, with the good repeatability in different experimental conditions. The fi/s between other three components with TIIA was accurate and reliable, and there was no significant difference between QAMS and ESM method. Besides, the relative retention value method could be used to accurately locate the measuring component. The relative retention values of DTI, TI, and CT to TIIA were 0.314, 0.518, and 0.561, respectively. The contents of three kinds of S. miltiorrhiza preparations showed there were some differences in the intrinsic quality of the products from different factories, even from different batches of the same manufacturer. Conclusion: The QAMS method could be used for quality control about related preparations of S. miltiorrhiza and provide references for quality evaluation of other preparations. At the same time, the above three kinds of traditional Chinese medicine preparations should be further strengthened the control and supervision.
ABSTRACT
OBJECTIVE: To predict the potential for drug-drug interactions of Salvia miltiorrhiza Bge. preparations with other-drugs, the effects of 11 active components of Salvia miltiorrhiza Bge. on the catalytic activities of 5 drug-metabolizing enzymes were assessed. METHODS: Human liver microsome incubation experiment was carried out to assay these components on the catalytic activities of these enzymes. Phenacetin, tolbutamide, 5-mephenytoin, dextromethorphane and midazolam were used as the substrates of (CYP1A2, CYP2C9, CYP2C19, CYP2D6 and CYP3A4, respectively. After the incubation, the metabolite of each substrate was quantified by LC-MS/MS method. The inhibitions of five main drug metabolism enzymes were calculated by comparing the formation velocities of metabolites with or without Salvia miltiorrhiza Bge. component. RESULTS: Some components of Salvia miltiorrhiza Bge. showed obvious inhibiton on the catalytic activities of CYPs, especially dihydrotanshinone I showed strong inhibiton on CYP3A4 with IC50 value of 0.367 μmol·L-1. CONCLUSION: Salvia miltiorrhiza Bge. preparation with high content of dihydrotanshinone I may cause drug-drug interactions. To predict the interaction precisely, the content of dihydrotanshinone I as well as its in vivo process need a further study.
ABSTRACT
Tanshinones are a large class of hydrophobic natural products isolated from traditional Chinese herb Salvia miltiorrhiza Bunge and related plants. Tanshinones include tanshinone ⅡA, cryptotanshinone, tanshinone I, dihydrotanshinone, and so on. Both the tanshinones and the salvianolic acids have been identi-fied as the characteristic and main active ingredients of Salvia miltiorrhiza Bunge with cardiovascular protective activities. Ac-cumulated data in recent years have revealed that tanshinones possess remarkable anti-cancer activities both in vivo and in vitro. In this review, we summarize the latest progress of the an-ti-cancer effect and the underlying mechanisms of tanshinones, particularly with emphasis on tanshinone ⅡA, which might pro-vide reference for the further research and development of these compounds.
ABSTRACT
Objective: To establish a UPLC-MS/MS method for determining the plasma concentration of dihydrotanshinone I, tanshinone I, cryptotanshinone, and tanshinone IIA in rats, and to study the pharmacokinetics of Salvia miltiorrhiza diterpene quinones composition (SMDQC) and its solid dispersion micro-pellets (SDMP). Methods: Sprague-Dawley rats were ig administered with SMDQC and its SDMP, respectively. Then the blood samples were obtained at different time points. Electrospray ionization (ESI) source was applied and operated in the positive ion mode. The plasma concentration of dihydrotanshinone I, tanshinone I, cryptotanshinone, and tanshinone IIA was then detected by UPLC-MS/MS, and the pharmcoknetic parameters were calculated by DAS 2.1.1 program. Results: The RSDs of intra- and inter-day precisions of all analytes were less than 14.6%, and the average recoveries of the four active constituents were more than 74.49%. The pharmacokinetic results showed that after ig administration of SDMP, Cmax and AUC0-∞ of the four active constituents increased significantly compared with those of SMDQC. Conclusion: The method has the high sensitivity and selectivity, and proves to be suitable for the pharmacokinetic study of SMDQC and its SDMP. The results show that the SDMP could enhance the solubility of SMDQC to improve its absorption. The relative bioavailability of the four representative constituents is 138%-204% of the crude drug.