AIM:This study was designed to evaluate the anti-cancer actions of tanshinone I and tanshinone IIA,and six derivatives of tanshinone IIA on normal and cancerous colon cells.Structure activity relationship(SAR) analysi...AIM:This study was designed to evaluate the anti-cancer actions of tanshinone I and tanshinone IIA,and six derivatives of tanshinone IIA on normal and cancerous colon cells.Structure activity relationship(SAR) analysis was conducted to delineate the significance of the structural modifications of tanshinones for improved anti-cancer action.METHOD:Tanshinone derivatives were designed and synthesized according to the literature.The cytotoxicity of different compounds on colon cancer cells was determined by the MTT assay.Apoptotic activity of the tanshinones was measured by flow cytometry(FCM).RESULTS:Tanshinone I and tanshinone IIA both exhibited significant cytotoxicity on colon cancer cells.They are more effective in p53+/+ colon cancer cell line.It was also noted that the anti-cancer activity of tanshinone I was more potent and selective.Two of the derivatives of tanshinone IIA(N1 and N2) also exhibited cytotoxicity on colon cancer cells.CONCLUSIONS:The anti-colon cancer activity of tanshinone I was more potent and selective than tanshinone IIA,and is p53 dependent.The derivatives obtained by structural modifications of tanshinone IIA exhibited lower cytotoxicity on both normal and colon cancer cells.From steric and electronic characteristics point of view,it was concluded that structural modifications of ring A and furan or dihydrofuran ring D on the basic structure of tanshinones influences the activity.An increase of the delocalization of the A and B rings could enhance the cytotoxicity of such compounds,while a non-planar and small sized D ring region would provide improved anti-cancer activity.展开更多
Total tanshinones are lipophilic active constituents extracted from Salvia miltiorrhiza Bge.Tanshinone ⅡA and cryptotanshinone are the major components in total tanshinones.However, the bioavailability of both compou...Total tanshinones are lipophilic active constituents extracted from Salvia miltiorrhiza Bge.Tanshinone ⅡA and cryptotanshinone are the major components in total tanshinones.However, the bioavailability of both compounds is low due to poor water solubility. To enhance the solubility and dissolution rate of tanshinone ⅡA, cryptotanshinone and total tanshinones,three common used hydrophilic carriers including PEG 6000, poloxamer 188 and PVP K30 were used to prepare the solid dispersions at different ratios, respectively. The solid dispersions were characterised by scanning electron microscopy(SEM), differential scanning calorimetry(DSC) and Fourier transform infrared spectroscopy(FTIR). The results of powder X-ray diffraction confirmed the microcrystal state of total tanshinones in solid dispersions and no chemical interaction between total tanshinones and carriers was observed in FTIR spectra. The solubility and dissolution rate of tanshinone ⅡA and cryptotanshinone were significantly increased in all solid dispersions. Regarding tanshinone ⅡA, the solubility and dissolution rate of in solid dispersions prepared with poloxamer 188 were significantly higher than that with PEG 6000 and PVP K30. The higher solubility and dissolution rate of cryptotanshinone were obtained in solid dispersion of PVP K30 than that of PEG 6000 solid dispersions but no significant difference from poloxamer 188 solid dispersions. The results indicate that the superior carrier for preparation of tanshinone ⅡA and total tanshinones solid dispersions is poloxamer 188, and that for cryptotanshinone is PVP K30.展开更多
The spectra of four tanshinones in the potentiostatic reduction process, including tanshinone I, dihydrotanshinone I, tanshinone IIA and cryptotanshinone, were investigated using spectroelectrochemical cell and UV spe...The spectra of four tanshinones in the potentiostatic reduction process, including tanshinone I, dihydrotanshinone I, tanshinone IIA and cryptotanshinone, were investigated using spectroelectrochemical cell and UV spectrophotometer. Their cyclic voltammograms （CVs） were recorded with a glassy carbon electrode （GCE）. The experiment results show that the antioxidant activity of these tanshinones, in the structure, where A, B and C rings connect through a single double bond, is weaker than that where A ring does not have double bond. Moreover, the increasing angle strain in the reduction process could enhance the antioxidant activity. In summary, the rank of antioxidant activities of these tanshinones, from weak to strong, is tanshinone I, dihydrotanshinone I, tanshinone IIA and cryptotanshinone.展开更多
[Objectives] To study the anti-inflammatory activity and mechanism of tanshinone I,cryptotanshinone and 15,16-dihydrotanshinone I on HAECs induced by TNF-α. [Methods]Vitamin E was used as a positive control and TNF-...[Objectives] To study the anti-inflammatory activity and mechanism of tanshinone I,cryptotanshinone and 15,16-dihydrotanshinone I on HAECs induced by TNF-α. [Methods]Vitamin E was used as a positive control and TNF-α-induced human aortic endothelial cells( HAECs) were selected as the inflammation model cells. The m RNA levels of IL-6,ICAM-1,VCAM-1,and NF-κB were analyzed by quantitative RT-PCR. The protein expression of NF-κB,ICAM-1,VCAM-1 and phosphorylation of ERK1/2 were determined by Western blot. The adhesion of U937 to HAECs was assessed by BCECF/AM labeling assay. [Results] TNF-α-induced over-expression of IL-6,NF-κB,ICAM-1,and VCAM-1 in HAECs were down-regulated by tanshinone I( TAN),cryptotanshinone( CPT) and 15,16-dihydrotanshinone I( DHT) both in m RNA and protein levels,respectively. Meanwhile 15,16-dihydrotanshinone I and cryptotanshinone could inhibit phosphorylation of ERK1/2 and tanshinone I inhibited U937 adhesion to HAECs significantly. [Conclusions] Tanshinones could inhibit TNF-α induced inflammatory responses on HAECs and the mechanism might be related to inhibition of phosphorylation of ERK1/2 and blocking NF-κB signaling pathway.展开更多
11β-Hydroxysteroid dehydrogenase 1(11β-HSD1)represents a promising drug target for metabolic syndrome,includ-ing obesity and type 2 diabetes.Our initial screen of a collection of natural products from Danshen led to...11β-Hydroxysteroid dehydrogenase 1(11β-HSD1)represents a promising drug target for metabolic syndrome,includ-ing obesity and type 2 diabetes.Our initial screen of a collection of natural products from Danshen led to the identi-fication of tanshinones as the potent and selective 11β-HSD1 inhibitors.To improve the druggability and explore the structure-activity relationships(SARs),more than 40 derivatives have been designed and synthesized using tanshinone IIA and cryptotanshinone as the starting materials.More than 10 derivatives exhibited potent in vitro 11β-HSD1 inhibitory activity and good selectivity over 11β-HSD2 across human and mouse species.Based on the biological results,SARs were further discussed,which was also partially rationalized by a molecular docking model of 1 bound to the 11β-HSD1.Remarkably,compounds 1,17 and 30 significantly inhibited 11β-HSD1 in 3T3-L1 adipocyte and in livers of ob/ob mice,which merits further investigations as anti-diabetic agents.This study not only provides a series of novel selective 11β-HSD1 inhibitors with promising therapeutic potentials in metabolic syndromes,but also expands the boundaries of the chemical and biological spaces of tanshinones.展开更多
[ Objective ] This study was conducted to improve quality standard of Compound Danshen tablets. [ Method] Tanshinones in Compound Danshen tab- lets were determined by HPLC method. [ Result] A good linear relationship ...[ Objective ] This study was conducted to improve quality standard of Compound Danshen tablets. [ Method] Tanshinones in Compound Danshen tab- lets were determined by HPLC method. [ Result] A good linear relationship was found in the range of 0.10 -0.50 μg, and the average recovery rate was 100.59% （RSD = 1.38% ）. [ Conclusion] The method is simple, rapid and reproducible, and could be used as a method for quality control of Compound Danshen Tablets.展开更多
Objective： To extract three kinds of tanshinones from the root of Salvia miltiorrhiza Bunge by CO2-supercdtical fluid extraction technology with different entrainers at different flow rates, and to investigate the ef...Objective： To extract three kinds of tanshinones from the root of Salvia miltiorrhiza Bunge by CO2-supercdtical fluid extraction technology with different entrainers at different flow rates, and to investigate the effects of different entrainers. Methods： Three kinds of tanshinones were extracted at the optimal operation condition, and the massconcentrafion of three kinds of tanshinones in the extracts was determined by HPLC. Results： Among the three enu＇ainers, the extracting effects of ethanol is the best, for the stronger polarity, followed by ethanol and normal octane. Conclusion： To increase the extracting rate of three kinds of tanshinones by CO2- supewrifical fluid extraction technics, it is essential to use polar solvent as entrainer.展开更多
OBJECTIVE:To identify the active anti-tumor constituents in the extract from Danshen(Radix Salviae Miltiorrhizae) and investigate the mechanisms underlying the actions.METHODS:First,we introduced a two-step counter-cu...OBJECTIVE:To identify the active anti-tumor constituents in the extract from Danshen(Radix Salviae Miltiorrhizae) and investigate the mechanisms underlying the actions.METHODS:First,we introduced a two-step counter-current chromatography to extract the therapeutically active diterpenoid,tanshinone from Danshen(Radix Salviae Miltiorrhizae).The cholecystokinin(CCK-8) method was used to evaluate the inhibitory effect of diterpenoid tanshinone in liver cancer QGY-7703,lung cancer PC9,lung cancer A549,gastric cancer MKN-45,gastric cancer HGC-27,colon cancer HCT116,myeloma cell U266/RPMI8226,and human breast cancer MCF-7 in vitro.Fluorescence staining was used to observe the cytotoxicity ofditerpenoid tanshinone on PC9 cells.The Western blot was used to detect apoptosis-related protein poly ADP-ribose polymerase(PARP),cysteinyl aspartate specific proteinase3/9(caspase3/9),and cleaved-cysteinyl aspartate specific proteinase3/9(cleaved-caspase3/9).The endoplasmic reticulum stress-related activating transcription factor 4(ATF4),phosphorylated eukaryotic initiation factor 2α(p-e IF2α),and phosphorylated jun amino-terminal kinase(p-JNK),and caspase-12 were also analyzed using the Western blot.RESULTS:Diterpenoid tanshinone inhibited the nine human tumor cell lines,with an IC50 of4.37-29 μg/m L,with the PC9 and MCF-7 displaying the lowest values.Fluorescence staining showed a lethal effect of diterpenoid tanshinone on PC9 cells.The Western blot showed that the expression of caspase3/9 protein and ATF-4 protein decreased gradually.However,the PARP,cleaved-caspase 3/9and the expression of p-e IF2 α,P-JNK,and caspase-12 increased gradually,in a dose-dependent fashion.CONCLUSION:We successfully introduced a two-step counter-current chromatography method to extract diterpenoid tanshinone,and demonstrated its antitumor activity.Diterpenoid tanshinone can induce apoptosis in nine human cancer cell lines.展开更多
The effects of LaCl3 on the growth,photosynthetic gas-exchange characteristics,chlorophyll fluorescence,and the accumulation of tanshinones and salvianolic acids in Salvia miltiorrhiza seedlings were investigated. The...The effects of LaCl3 on the growth,photosynthetic gas-exchange characteristics,chlorophyll fluorescence,and the accumulation of tanshinones and salvianolic acids in Salvia miltiorrhiza seedlings were investigated. The results showed that the increase in photosynthesis induced by LaCl3 might be attributed to the enhanced stomatal conductance of the leaves and the increased level of the photochemical efficiency of PS II. The accumulation of tanshinone IIA and cryptotanshinone was markedly increased with the application of LaCl3 at 20 and 60 mg/L,while tanshinone I was only slightly increased. The content of salvianolic acid B was,however,decreased with the treatment of LaCl3 at 200 mg/L.展开更多
The reaction of cryptotanshinone and tanshinone IIA with several biogenic amine metabolites involved in the pathogenic pathways of HE were investigated and eight 1,2,3,4- tetrahydrophenanthrene derivatives, 2-6 and 8-...The reaction of cryptotanshinone and tanshinone IIA with several biogenic amine metabolites involved in the pathogenic pathways of HE were investigated and eight 1,2,3,4- tetrahydrophenanthrene derivatives, 2-6 and 8-10, were obtained. The probable mechanism on reaction was discussed.展开更多
Plants have mechanisms to transport secondary metabolites from where they are biosynthesized to the sites where they function,or to sites such as the vacuole for detoxification.However,current research has mainly focu...Plants have mechanisms to transport secondary metabolites from where they are biosynthesized to the sites where they function,or to sites such as the vacuole for detoxification.However,current research has mainly focused on metabolite biosynthesis and regulation,and little is known about their transport.Tanshinone,a class diterpenoid with medicinal properties,is biosynthesized in the periderm of Salvia miltiorrhiza roots.Here,we discovered that tanshinone can be transported out of peridermal cells and secreted into the soil environment and that the ABC transporter SmABCG1 is involved in the efflux of tanshinoneⅡA and tanshinoneⅠ.The SmABCG1 gene is adjacent to the diterpene biosynthesis gene cluster in the S.miltiorrhiza genome.The temporal–spatial expression pattern of SmABCG1 is consistent with tanshinone accumulation profiles.SmABCG1 is located on the plasma membrane and preferentially accumulates in the peridermal cells of S.miltiorrhiza roots.Heterologous expression in Xenopus laevis oocytes demonstrated that SmABCG1 can export tanshinoneⅡA and tanshinoneⅠ.CRISPR/Cas9-mediated mutagenesis of SmABCG1 in S.miltiorrhiza hairy roots resulted in a significant decrease in tanshinone contents in both hairy roots and the culture medium,whereas overexpression of this gene resulted in increased tanshinone contents.CYP76AH3 transcript levels increased in hairy roots overexpressing SmABCG1 and decreased in knockout lines,suggesting that SmABCG1 may affect the expression of CYP76AH3,indirectly regulating tanshinone biosynthesis.Finally,tanshinoneⅡA showed cytotoxicity to Arabidopsis roots.These findings offer new perspectives on plant diterpenoid transport and provide a new genetic tool for metabolic engineering and synthetic biology research.展开更多
Objective:Salvia miltiorrhiza is a valuable herbal medicine with tanshinone and phenolic acid as the main biological active ingredients.The biosynthetic regulation of these bioactive compounds is controlled by a set o...Objective:Salvia miltiorrhiza is a valuable herbal medicine with tanshinone and phenolic acid as the main biological active ingredients.The biosynthetic regulation of these bioactive compounds is controlled by a set of transcription factors(TFs).The basic helix-loop-helix(bHLH)transcription factor plays an important role in various physiological and biochemical processes in plants.However,research on bHLH TFs regulating phenolic acid or tanshinone biosynthesis in S.miltiorrhiza is limited.Methods:qRT-PCR was used for gene expression analysis.The subcellular localization of SmbHLH92 was detected by SmbHLHg2-GFP transient transformation into tobacco leaves,and its fluorescence was observed using a confocal laser scanning microscope.The transcriptional activity of SmbHLH92 was confirmed in the AH109 yeast strain.RNA interference hairy roots of SmbHLH92-RNAi transgenic lines were obtained through Agrobacterium-mediated genetic transformation.Ultra performance liquid chromatography(UPLC)was used to detect the changes of phenolic acids and tanshinones.Results:SmbHLH92 is a bHLH transcription factor that is highly expressed in the root and phloem of S.miltiorrhiza.The subcellular localization and transcriptional activity of SmbHLH92 indicated that SmbHLH92 was located in the nucleus and may be a transcription factor.RNA interference(RNAi)of SmbHLH92 in hairy roots of S.miltiorrhiza significantly increased the accumulation of phenolic acid and tanshinone.Quantitative RT-PCR(RT-qPCR)analysis showed the transcription level of genes encoding the key enzymes involved in the phenolic acid and tanshinone biosynthetic pathways was increased in the hairy roots of the SmbHLH92-RNAi transgenic line,comparing with the control line.Conclusion:These data indicate that SmbHLH92 is a negative regulator involved in the regulation of phenolic acid and tanshinone biosynthesis in S.miltiorrhiza.展开更多
To enhance the structural diversity of tanshinones and provide more derivatives for the biological studies, a one-pot combinatorial modification strategy was performed on total tanshinones. Six new quinoxlinetanshinon...To enhance the structural diversity of tanshinones and provide more derivatives for the biological studies, a one-pot combinatorial modification strategy was performed on total tanshinones. Six new quinoxlinetanshinones 1 --6 were subsequently isolated from the combinatorial modified semi-synthetic mixture. The structures were elu- cidated by spectroscopic analysis in combination with single-crystal X-ray diffraction. These quinoxlinetanshinones demonstrated strong DNA binding properties. Rapid synthesis of new quinoxlinetanshinones with significant bio- logical activity highlights the great potential of one-pot combinatorial modification for the diversification of natural products.展开更多
Objective:Tanshinone ⅡA,one of the most abundant liposoluble components isolated from the traditional Chinese medicine Salvia miltiorrhiza,exhibits significant biological activities in anti-inflammatory,antibacterial...Objective:Tanshinone ⅡA,one of the most abundant liposoluble components isolated from the traditional Chinese medicine Salvia miltiorrhiza,exhibits significant biological activities in anti-inflammatory,antibacterial,and antitumor eff ects.This study aims to systematically explore the mechanism of Tanshinone ⅡA through bioinformatics.Methods:We utilized the TCMSP database to retrieve the oral bioavailability(OB)and drug-likeness(DL)of Tanshinone ⅡA.The gene chip numbered GSE85871 was downloaded from the GEO database,and diff erential genes were analyzed using R language to identify potential targets of Tanshinone ⅡA.After obtaining these targets,GO analysis and KEGG pathway analysis were performed using the DAVID 6.8 database.Diseases related to Tanshinone ⅡA were explored through the CTD database.Finally,Cytoscape was employed to construct a visual network of multiple targets,pathways,and diseases associated with Tanshinone ⅡA.Results:Tanshinone ⅡA demonstrated good drug effi cacy with an OB value of 49.89%and a DL value of 0.4.A total of 132 potential targets were identifi ed,primarily exhibiting gene co-expression and physical interaction in the PPI network.These targets were enriched in biological processes and pathways such as ovarian steroidogenesis,cell cycle,and steroid hormone biosynthesis.Tanshinone ⅡA was found to be relevant in the treatment of diseases including breast tumors,hypertension,atherosclerosis,gliomas,vascular system injuries,left ventricular hypertrophy,leukemia,and hearing loss.Conclusion:Utilizing bioinformatics approaches,we systematically analyzed the possible molecular mechanisms of Tanshinone ⅡA,providing potential targets and insights into its pharmacological mechanisms and treatment strategies.展开更多
Objective:Salvia miltiorrhiza is widely used in traditional Chinese medicine for treating cardiovascular and cerebrovascular diseases,with tanshinones being its major active components.This study aims to systematicall...Objective:Salvia miltiorrhiza is widely used in traditional Chinese medicine for treating cardiovascular and cerebrovascular diseases,with tanshinones being its major active components.This study aims to systematically elucidate the core transcriptional circuitry controlling tanshinone production,thereby establishing a mechanistic framework to optimize phytochemical yield and advance sustainable cultivation strategies for this pharmaceutically vital species.Methods:Transcriptome profiling revealed that the transcription factor SmWRKY69 is specifically expressed in the root periderm of S.miltiorrhiza.DNA affinity purification sequencing(DAPseq)was used to identify its potential target genes,and cis-element analysis predicted W-box motifs in the promoters of SmCPS1 and SmKSL1.Yeast one-hybrid(Y1H)assays were employed to validate its regulatory interactions with candidate gene promoters.Results:SmWRKY69 was found to directly bind to the promoters of SmCPS1 and SmKSL1,key genes in the tanshinone biosynthetic pathway,through W-box elements,indicating its role as a transcriptional regulator.Conclusion:SmWRKY69 regulates tanshinone biosynthesis by directly targeting SmCPS1 and SmKSL1,providing a valuable genetic target for metabolic engineering to enhance the therapeutic quality of S.miltiorrhiza.展开更多
A method of gradient-elution HPLC with UV detection was developed for theanalysis of nine major constituents of Salvia species, a commonly used TCM herb, namely danshensu,protocatechuic acid, protocatechualdehyde, sal...A method of gradient-elution HPLC with UV detection was developed for theanalysis of nine major constituents of Salvia species, a commonly used TCM herb, namely danshensu,protocatechuic acid, protocatechualdehyde, salviano-lic acid B, methyltanshinone, dihydrotanshinone,cryptotanshinone, tanshinoneⅠand tanshinoneⅡ_A. In the present study, a Shimadzu CLC-ODS column(150 mm x 6 mm, 5 μm) was utilized and 0.5% formic acid (A) and acetonitrile (B) were used forgradient elution at a total flow rate of 0.8 mL· min^(-1). All calibration curves showed goodlinear regression ( r > 0.999) within test ranges. Extraction was conducted by refluxing methanol(10 mL) with dried herb (0.5 g) for 1.0 h.The assay was simple, convenient and reproducible. Theproposed method was successfully applied to the determination of nine major constituents in thirteenSalvia. species and the results showed that the contents of Salvia components vary in differentspecies and origin. Tanshinone was hardly detected in S. yunnanensis and S. prionitis, thereforethey are not suitable for clinical use as Danshen.展开更多
[Objective]The aim was to extract tanshinone from Salvia miltiorrhiza Bge. dregs and to determine tanshinone components. [Method]Organic solvent method was adopted to extract tanshinone from S. miltiorrhiza dregs and ...[Objective]The aim was to extract tanshinone from Salvia miltiorrhiza Bge. dregs and to determine tanshinone components. [Method]Organic solvent method was adopted to extract tanshinone from S. miltiorrhiza dregs and TLC was used to determine the optimum extraction solvent. The components of tanshinone were measured with HPLC. [Result]Ether was the best solvent to extract tanshinone from S. miltiorrhiza dregs. After water immersion,dry dregs of S. miltiorrhiza and Panax notoginseng were extracted with ethanol to obtain fat-soluble extracts. Then with ether as the solvent for Soxhlet extraction,the yield of crude tanshinone was 2.17%. The HPLC detection showed that the contents of tanshinone Ⅱ A,methylene tanshinquinone,cryptotanshinone,tanshinone Ⅰ were 3.62%,1.02%,2.56%,2.75% respectively. [Conclusion]The components of tanshinone in S. miltiorrhiza dregs were basically the same as tanshinone in medicine S. miltiorrhiza. S. miltiorrhiza dregs could be used as a kind of tanshinone resource,which has the value of development and utilization.展开更多
Multidrug resistance(MDR)develops during chemotherapy in nearly all colorectal cancerpatients.It is envisaged that reversal of MDR plays a pivotal role in the success of chemotherapy.This study investigated thepotenti...Multidrug resistance(MDR)develops during chemotherapy in nearly all colorectal cancerpatients.It is envisaged that reversal of MDR plays a pivotal role in the success of chemotherapy.This study investigated thepotential pharmacological action in reversing MDR in colon cancer cells by the two most potent tanshinones,namely cryptotanshinone and dihydrotanshinone.They targeted two common MDR mechanisms,including overexpression of P-glycoprotein(P-gp)and suppression of apoptosis.Using a bi-directional transport assay,the two tanshinones decreased P-gp-mediated digoxin effluxin Caco-2 cells.They also potentiated the cytotoxicities of doxorubicin and irinotecan in P-gp overexpressing SW620Ad300 cells via increased intracellular accumulation of both anti-cancer drugs,as a result of down-regulation of P-gp mRNA and protein levels as well as inhibition of P-gp ATPase activity.In addition,the level of apoptosis was also found to be relatively suppressed in SW620Ad300 cells as compared with the parental SW620 cells.Interestingly,although cryptotanshinone and dihydrotanshinone induced less apoptosis in SW620Ad300 cells as compared to their parental cells,they produced more autophagic cell death in these MDR cells.In this regard,the drug resistant SW620Ad300 cells were more prone to cell death in response to the anti-cancer action of thetwo tanshinones.Furthermore,the cytotoxic action of the two tanshinones was shown to be p53-independent,further demonstrated theirunique anti-cancer activities in overcoming drug resistance due to the reduction of p53 expression together with a decrease of apoptosis in colon cancer cells.Taken together,the current findings indicate a great potential for cryptotanshinone and dihydrotanshinone against MDR colon cancer cells,in spite of P-gp overexpression and suppression of apoptosis.They are promising candidates to be developed as therapeutic agents and/or as an adjuvant therapy for colorectal cancer,especially for patients with MDR cancer types.展开更多
基金supported by the National Major Scientific and Technological Special Project for"Significant New Drugs Development"(No.2011ZX09401-028)
文摘AIM:This study was designed to evaluate the anti-cancer actions of tanshinone I and tanshinone IIA,and six derivatives of tanshinone IIA on normal and cancerous colon cells.Structure activity relationship(SAR) analysis was conducted to delineate the significance of the structural modifications of tanshinones for improved anti-cancer action.METHOD:Tanshinone derivatives were designed and synthesized according to the literature.The cytotoxicity of different compounds on colon cancer cells was determined by the MTT assay.Apoptotic activity of the tanshinones was measured by flow cytometry(FCM).RESULTS:Tanshinone I and tanshinone IIA both exhibited significant cytotoxicity on colon cancer cells.They are more effective in p53+/+ colon cancer cell line.It was also noted that the anti-cancer activity of tanshinone I was more potent and selective.Two of the derivatives of tanshinone IIA(N1 and N2) also exhibited cytotoxicity on colon cancer cells.CONCLUSIONS:The anti-colon cancer activity of tanshinone I was more potent and selective than tanshinone IIA,and is p53 dependent.The derivatives obtained by structural modifications of tanshinone IIA exhibited lower cytotoxicity on both normal and colon cancer cells.From steric and electronic characteristics point of view,it was concluded that structural modifications of ring A and furan or dihydrofuran ring D on the basic structure of tanshinones influences the activity.An increase of the delocalization of the A and B rings could enhance the cytotoxicity of such compounds,while a non-planar and small sized D ring region would provide improved anti-cancer activity.
文摘Total tanshinones are lipophilic active constituents extracted from Salvia miltiorrhiza Bge.Tanshinone ⅡA and cryptotanshinone are the major components in total tanshinones.However, the bioavailability of both compounds is low due to poor water solubility. To enhance the solubility and dissolution rate of tanshinone ⅡA, cryptotanshinone and total tanshinones,three common used hydrophilic carriers including PEG 6000, poloxamer 188 and PVP K30 were used to prepare the solid dispersions at different ratios, respectively. The solid dispersions were characterised by scanning electron microscopy(SEM), differential scanning calorimetry(DSC) and Fourier transform infrared spectroscopy(FTIR). The results of powder X-ray diffraction confirmed the microcrystal state of total tanshinones in solid dispersions and no chemical interaction between total tanshinones and carriers was observed in FTIR spectra. The solubility and dissolution rate of tanshinone ⅡA and cryptotanshinone were significantly increased in all solid dispersions. Regarding tanshinone ⅡA, the solubility and dissolution rate of in solid dispersions prepared with poloxamer 188 were significantly higher than that with PEG 6000 and PVP K30. The higher solubility and dissolution rate of cryptotanshinone were obtained in solid dispersion of PVP K30 than that of PEG 6000 solid dispersions but no significant difference from poloxamer 188 solid dispersions. The results indicate that the superior carrier for preparation of tanshinone ⅡA and total tanshinones solid dispersions is poloxamer 188, and that for cryptotanshinone is PVP K30.
文摘The spectra of four tanshinones in the potentiostatic reduction process, including tanshinone I, dihydrotanshinone I, tanshinone IIA and cryptotanshinone, were investigated using spectroelectrochemical cell and UV spectrophotometer. Their cyclic voltammograms （CVs） were recorded with a glassy carbon electrode （GCE）. The experiment results show that the antioxidant activity of these tanshinones, in the structure, where A, B and C rings connect through a single double bond, is weaker than that where A ring does not have double bond. Moreover, the increasing angle strain in the reduction process could enhance the antioxidant activity. In summary, the rank of antioxidant activities of these tanshinones, from weak to strong, is tanshinone I, dihydrotanshinone I, tanshinone IIA and cryptotanshinone.
基金Supported by the National Natural Science Foundation of China(81274031)
文摘[Objectives] To study the anti-inflammatory activity and mechanism of tanshinone I,cryptotanshinone and 15,16-dihydrotanshinone I on HAECs induced by TNF-α. [Methods]Vitamin E was used as a positive control and TNF-α-induced human aortic endothelial cells( HAECs) were selected as the inflammation model cells. The m RNA levels of IL-6,ICAM-1,VCAM-1,and NF-κB were analyzed by quantitative RT-PCR. The protein expression of NF-κB,ICAM-1,VCAM-1 and phosphorylation of ERK1/2 were determined by Western blot. The adhesion of U937 to HAECs was assessed by BCECF/AM labeling assay. [Results] TNF-α-induced over-expression of IL-6,NF-κB,ICAM-1,and VCAM-1 in HAECs were down-regulated by tanshinone I( TAN),cryptotanshinone( CPT) and 15,16-dihydrotanshinone I( DHT) both in m RNA and protein levels,respectively. Meanwhile 15,16-dihydrotanshinone I and cryptotanshinone could inhibit phosphorylation of ERK1/2 and tanshinone I inhibited U937 adhesion to HAECs significantly. [Conclusions] Tanshinones could inhibit TNF-α induced inflammatory responses on HAECs and the mechanism might be related to inhibition of phosphorylation of ERK1/2 and blocking NF-κB signaling pathway.
基金the National Natural Science Foundation of China (No.U1502223)Hunan Provincial Key Research and Development Project (Grant No.2021WK2005 to X.Deng)+1 种基金Natural Science Foundation of Hunan Province (Grant No.2021JJ30894 to X.Deng)the open fund of State Key Laboratory of Phytochemistry and Plant Resource in West China (Grant No.P2020-KF03).
文摘11β-Hydroxysteroid dehydrogenase 1(11β-HSD1)represents a promising drug target for metabolic syndrome,includ-ing obesity and type 2 diabetes.Our initial screen of a collection of natural products from Danshen led to the identi-fication of tanshinones as the potent and selective 11β-HSD1 inhibitors.To improve the druggability and explore the structure-activity relationships(SARs),more than 40 derivatives have been designed and synthesized using tanshinone IIA and cryptotanshinone as the starting materials.More than 10 derivatives exhibited potent in vitro 11β-HSD1 inhibitory activity and good selectivity over 11β-HSD2 across human and mouse species.Based on the biological results,SARs were further discussed,which was also partially rationalized by a molecular docking model of 1 bound to the 11β-HSD1.Remarkably,compounds 1,17 and 30 significantly inhibited 11β-HSD1 in 3T3-L1 adipocyte and in livers of ob/ob mice,which merits further investigations as anti-diabetic agents.This study not only provides a series of novel selective 11β-HSD1 inhibitors with promising therapeutic potentials in metabolic syndromes,but also expands the boundaries of the chemical and biological spaces of tanshinones.
文摘[ Objective ] This study was conducted to improve quality standard of Compound Danshen tablets. [ Method] Tanshinones in Compound Danshen tab- lets were determined by HPLC method. [ Result] A good linear relationship was found in the range of 0.10 -0.50 μg, and the average recovery rate was 100.59% （RSD = 1.38% ）. [ Conclusion] The method is simple, rapid and reproducible, and could be used as a method for quality control of Compound Danshen Tablets.
文摘Objective： To extract three kinds of tanshinones from the root of Salvia miltiorrhiza Bunge by CO2-supercdtical fluid extraction technology with different entrainers at different flow rates, and to investigate the effects of different entrainers. Methods： Three kinds of tanshinones were extracted at the optimal operation condition, and the massconcentrafion of three kinds of tanshinones in the extracts was determined by HPLC. Results： Among the three enu＇ainers, the extracting effects of ethanol is the best, for the stronger polarity, followed by ethanol and normal octane. Conclusion： To increase the extracting rate of three kinds of tanshinones by CO2- supewrifical fluid extraction technics, it is essential to use polar solvent as entrainer.
基金Supported by Key Projects of Zhejiang Province Science and Technology(Arsenic Trioxide Injection Associate with Tanshinone Treat the Hepatocarcinoma of Qi-Stagnancy and Blood Stasis,No.2012C13017-1)the Specialized Research Foundation for the Doctoral Program of Higher Education of China(the Mechanism of Jak-STAT3 Signaling Transduction in Anti-Hepatocarcinoma Associated with Arsenic Trioxide and Cryptotanshinone,No.20123322110001)
文摘OBJECTIVE:To identify the active anti-tumor constituents in the extract from Danshen(Radix Salviae Miltiorrhizae) and investigate the mechanisms underlying the actions.METHODS:First,we introduced a two-step counter-current chromatography to extract the therapeutically active diterpenoid,tanshinone from Danshen(Radix Salviae Miltiorrhizae).The cholecystokinin(CCK-8) method was used to evaluate the inhibitory effect of diterpenoid tanshinone in liver cancer QGY-7703,lung cancer PC9,lung cancer A549,gastric cancer MKN-45,gastric cancer HGC-27,colon cancer HCT116,myeloma cell U266/RPMI8226,and human breast cancer MCF-7 in vitro.Fluorescence staining was used to observe the cytotoxicity ofditerpenoid tanshinone on PC9 cells.The Western blot was used to detect apoptosis-related protein poly ADP-ribose polymerase(PARP),cysteinyl aspartate specific proteinase3/9(caspase3/9),and cleaved-cysteinyl aspartate specific proteinase3/9(cleaved-caspase3/9).The endoplasmic reticulum stress-related activating transcription factor 4(ATF4),phosphorylated eukaryotic initiation factor 2α(p-e IF2α),and phosphorylated jun amino-terminal kinase(p-JNK),and caspase-12 were also analyzed using the Western blot.RESULTS:Diterpenoid tanshinone inhibited the nine human tumor cell lines,with an IC50 of4.37-29 μg/m L,with the PC9 and MCF-7 displaying the lowest values.Fluorescence staining showed a lethal effect of diterpenoid tanshinone on PC9 cells.The Western blot showed that the expression of caspase3/9 protein and ATF-4 protein decreased gradually.However,the PARP,cleaved-caspase 3/9and the expression of p-e IF2 α,P-JNK,and caspase-12 increased gradually,in a dose-dependent fashion.CONCLUSION:We successfully introduced a two-step counter-current chromatography method to extract diterpenoid tanshinone,and demonstrated its antitumor activity.Diterpenoid tanshinone can induce apoptosis in nine human cancer cell lines.
基金Project supported by the National Natural Science Foundation of China (81072989)the Important National Science & Technology Specific Pro-jects (2009ZX09502-026, 2009ZX09301-005, 2009ZX09308-002)the Independent Research Projects of China Academy of Chinese Medi-cal Sciences (ZZ20090302)
文摘The effects of LaCl3 on the growth,photosynthetic gas-exchange characteristics,chlorophyll fluorescence,and the accumulation of tanshinones and salvianolic acids in Salvia miltiorrhiza seedlings were investigated. The results showed that the increase in photosynthesis induced by LaCl3 might be attributed to the enhanced stomatal conductance of the leaves and the increased level of the photochemical efficiency of PS II. The accumulation of tanshinone IIA and cryptotanshinone was markedly increased with the application of LaCl3 at 20 and 60 mg/L,while tanshinone I was only slightly increased. The content of salvianolic acid B was,however,decreased with the treatment of LaCl3 at 200 mg/L.
基金The program is sponsored by the Guangzhou City Science Foundation(2000-Z-021-01)Guangdong Provincial Science Foundation(2KM04103S).
文摘The reaction of cryptotanshinone and tanshinone IIA with several biogenic amine metabolites involved in the pathogenic pathways of HE were investigated and eight 1,2,3,4- tetrahydrophenanthrene derivatives, 2-6 and 8-10, were obtained. The probable mechanism on reaction was discussed.
基金financially supported by the National Key Research and Development Program of China(2022YFC3501700)the National Natural Science Foundation of China(82304662,82374159,32070327,32170402)+3 种基金the National Key Research and Development Program of China(2023YFC3504800)Chenguang project of Shanghai(23CGA52)Science and Technology Development Program of Shanghai University of Traditional Chinese Medicine(23KFL051)Shanghai Municipal Science and Technology Commission(23XD1423500)。
文摘Plants have mechanisms to transport secondary metabolites from where they are biosynthesized to the sites where they function,or to sites such as the vacuole for detoxification.However,current research has mainly focused on metabolite biosynthesis and regulation,and little is known about their transport.Tanshinone,a class diterpenoid with medicinal properties,is biosynthesized in the periderm of Salvia miltiorrhiza roots.Here,we discovered that tanshinone can be transported out of peridermal cells and secreted into the soil environment and that the ABC transporter SmABCG1 is involved in the efflux of tanshinoneⅡA and tanshinoneⅠ.The SmABCG1 gene is adjacent to the diterpene biosynthesis gene cluster in the S.miltiorrhiza genome.The temporal–spatial expression pattern of SmABCG1 is consistent with tanshinone accumulation profiles.SmABCG1 is located on the plasma membrane and preferentially accumulates in the peridermal cells of S.miltiorrhiza roots.Heterologous expression in Xenopus laevis oocytes demonstrated that SmABCG1 can export tanshinoneⅡA and tanshinoneⅠ.CRISPR/Cas9-mediated mutagenesis of SmABCG1 in S.miltiorrhiza hairy roots resulted in a significant decrease in tanshinone contents in both hairy roots and the culture medium,whereas overexpression of this gene resulted in increased tanshinone contents.CYP76AH3 transcript levels increased in hairy roots overexpressing SmABCG1 and decreased in knockout lines,suggesting that SmABCG1 may affect the expression of CYP76AH3,indirectly regulating tanshinone biosynthesis.Finally,tanshinoneⅡA showed cytotoxicity to Arabidopsis roots.These findings offer new perspectives on plant diterpenoid transport and provide a new genetic tool for metabolic engineering and synthetic biology research.
基金This work was supported by the National Natural Science Foundation(Grant No.31570302,81973422)Chinese Academy of Medical Sciences(CAMS)Innovation Fund for Medical Sciences(CIFMS,2016-I2M-3-016).
文摘Objective:Salvia miltiorrhiza is a valuable herbal medicine with tanshinone and phenolic acid as the main biological active ingredients.The biosynthetic regulation of these bioactive compounds is controlled by a set of transcription factors(TFs).The basic helix-loop-helix(bHLH)transcription factor plays an important role in various physiological and biochemical processes in plants.However,research on bHLH TFs regulating phenolic acid or tanshinone biosynthesis in S.miltiorrhiza is limited.Methods:qRT-PCR was used for gene expression analysis.The subcellular localization of SmbHLH92 was detected by SmbHLHg2-GFP transient transformation into tobacco leaves,and its fluorescence was observed using a confocal laser scanning microscope.The transcriptional activity of SmbHLH92 was confirmed in the AH109 yeast strain.RNA interference hairy roots of SmbHLH92-RNAi transgenic lines were obtained through Agrobacterium-mediated genetic transformation.Ultra performance liquid chromatography(UPLC)was used to detect the changes of phenolic acids and tanshinones.Results:SmbHLH92 is a bHLH transcription factor that is highly expressed in the root and phloem of S.miltiorrhiza.The subcellular localization and transcriptional activity of SmbHLH92 indicated that SmbHLH92 was located in the nucleus and may be a transcription factor.RNA interference(RNAi)of SmbHLH92 in hairy roots of S.miltiorrhiza significantly increased the accumulation of phenolic acid and tanshinone.Quantitative RT-PCR(RT-qPCR)analysis showed the transcription level of genes encoding the key enzymes involved in the phenolic acid and tanshinone biosynthetic pathways was increased in the hairy roots of the SmbHLH92-RNAi transgenic line,comparing with the control line.Conclusion:These data indicate that SmbHLH92 is a negative regulator involved in the regulation of phenolic acid and tanshinone biosynthesis in S.miltiorrhiza.
文摘To enhance the structural diversity of tanshinones and provide more derivatives for the biological studies, a one-pot combinatorial modification strategy was performed on total tanshinones. Six new quinoxlinetanshinones 1 --6 were subsequently isolated from the combinatorial modified semi-synthetic mixture. The structures were elu- cidated by spectroscopic analysis in combination with single-crystal X-ray diffraction. These quinoxlinetanshinones demonstrated strong DNA binding properties. Rapid synthesis of new quinoxlinetanshinones with significant bio- logical activity highlights the great potential of one-pot combinatorial modification for the diversification of natural products.
文摘Objective:Tanshinone ⅡA,one of the most abundant liposoluble components isolated from the traditional Chinese medicine Salvia miltiorrhiza,exhibits significant biological activities in anti-inflammatory,antibacterial,and antitumor eff ects.This study aims to systematically explore the mechanism of Tanshinone ⅡA through bioinformatics.Methods:We utilized the TCMSP database to retrieve the oral bioavailability(OB)and drug-likeness(DL)of Tanshinone ⅡA.The gene chip numbered GSE85871 was downloaded from the GEO database,and diff erential genes were analyzed using R language to identify potential targets of Tanshinone ⅡA.After obtaining these targets,GO analysis and KEGG pathway analysis were performed using the DAVID 6.8 database.Diseases related to Tanshinone ⅡA were explored through the CTD database.Finally,Cytoscape was employed to construct a visual network of multiple targets,pathways,and diseases associated with Tanshinone ⅡA.Results:Tanshinone ⅡA demonstrated good drug effi cacy with an OB value of 49.89%and a DL value of 0.4.A total of 132 potential targets were identifi ed,primarily exhibiting gene co-expression and physical interaction in the PPI network.These targets were enriched in biological processes and pathways such as ovarian steroidogenesis,cell cycle,and steroid hormone biosynthesis.Tanshinone ⅡA was found to be relevant in the treatment of diseases including breast tumors,hypertension,atherosclerosis,gliomas,vascular system injuries,left ventricular hypertrophy,leukemia,and hearing loss.Conclusion:Utilizing bioinformatics approaches,we systematically analyzed the possible molecular mechanisms of Tanshinone ⅡA,providing potential targets and insights into its pharmacological mechanisms and treatment strategies.
文摘Objective:Salvia miltiorrhiza is widely used in traditional Chinese medicine for treating cardiovascular and cerebrovascular diseases,with tanshinones being its major active components.This study aims to systematically elucidate the core transcriptional circuitry controlling tanshinone production,thereby establishing a mechanistic framework to optimize phytochemical yield and advance sustainable cultivation strategies for this pharmaceutically vital species.Methods:Transcriptome profiling revealed that the transcription factor SmWRKY69 is specifically expressed in the root periderm of S.miltiorrhiza.DNA affinity purification sequencing(DAPseq)was used to identify its potential target genes,and cis-element analysis predicted W-box motifs in the promoters of SmCPS1 and SmKSL1.Yeast one-hybrid(Y1H)assays were employed to validate its regulatory interactions with candidate gene promoters.Results:SmWRKY69 was found to directly bind to the promoters of SmCPS1 and SmKSL1,key genes in the tanshinone biosynthetic pathway,through W-box elements,indicating its role as a transcriptional regulator.Conclusion:SmWRKY69 regulates tanshinone biosynthesis by directly targeting SmCPS1 and SmKSL1,providing a valuable genetic target for metabolic engineering to enhance the therapeutic quality of S.miltiorrhiza.
文摘A method of gradient-elution HPLC with UV detection was developed for theanalysis of nine major constituents of Salvia species, a commonly used TCM herb, namely danshensu,protocatechuic acid, protocatechualdehyde, salviano-lic acid B, methyltanshinone, dihydrotanshinone,cryptotanshinone, tanshinoneⅠand tanshinoneⅡ_A. In the present study, a Shimadzu CLC-ODS column(150 mm x 6 mm, 5 μm) was utilized and 0.5% formic acid (A) and acetonitrile (B) were used forgradient elution at a total flow rate of 0.8 mL· min^(-1). All calibration curves showed goodlinear regression ( r > 0.999) within test ranges. Extraction was conducted by refluxing methanol(10 mL) with dried herb (0.5 g) for 1.0 h.The assay was simple, convenient and reproducible. Theproposed method was successfully applied to the determination of nine major constituents in thirteenSalvia. species and the results showed that the contents of Salvia components vary in differentspecies and origin. Tanshinone was hardly detected in S. yunnanensis and S. prionitis, thereforethey are not suitable for clinical use as Danshen.
基金Supported by National Natural Science Foundation of China(30470055)Innovation Team Project of Liaoning Education Department (2007T006)~~
文摘[Objective]The aim was to extract tanshinone from Salvia miltiorrhiza Bge. dregs and to determine tanshinone components. [Method]Organic solvent method was adopted to extract tanshinone from S. miltiorrhiza dregs and TLC was used to determine the optimum extraction solvent. The components of tanshinone were measured with HPLC. [Result]Ether was the best solvent to extract tanshinone from S. miltiorrhiza dregs. After water immersion,dry dregs of S. miltiorrhiza and Panax notoginseng were extracted with ethanol to obtain fat-soluble extracts. Then with ether as the solvent for Soxhlet extraction,the yield of crude tanshinone was 2.17%. The HPLC detection showed that the contents of tanshinone Ⅱ A,methylene tanshinquinone,cryptotanshinone,tanshinone Ⅰ were 3.62%,1.02%,2.56%,2.75% respectively. [Conclusion]The components of tanshinone in S. miltiorrhiza dregs were basically the same as tanshinone in medicine S. miltiorrhiza. S. miltiorrhiza dregs could be used as a kind of tanshinone resource,which has the value of development and utilization.
基金The project supported by the Hong Kong Research Grants Council and the Innovation Technology Commission
文摘Multidrug resistance(MDR)develops during chemotherapy in nearly all colorectal cancerpatients.It is envisaged that reversal of MDR plays a pivotal role in the success of chemotherapy.This study investigated thepotential pharmacological action in reversing MDR in colon cancer cells by the two most potent tanshinones,namely cryptotanshinone and dihydrotanshinone.They targeted two common MDR mechanisms,including overexpression of P-glycoprotein(P-gp)and suppression of apoptosis.Using a bi-directional transport assay,the two tanshinones decreased P-gp-mediated digoxin effluxin Caco-2 cells.They also potentiated the cytotoxicities of doxorubicin and irinotecan in P-gp overexpressing SW620Ad300 cells via increased intracellular accumulation of both anti-cancer drugs,as a result of down-regulation of P-gp mRNA and protein levels as well as inhibition of P-gp ATPase activity.In addition,the level of apoptosis was also found to be relatively suppressed in SW620Ad300 cells as compared with the parental SW620 cells.Interestingly,although cryptotanshinone and dihydrotanshinone induced less apoptosis in SW620Ad300 cells as compared to their parental cells,they produced more autophagic cell death in these MDR cells.In this regard,the drug resistant SW620Ad300 cells were more prone to cell death in response to the anti-cancer action of thetwo tanshinones.Furthermore,the cytotoxic action of the two tanshinones was shown to be p53-independent,further demonstrated theirunique anti-cancer activities in overcoming drug resistance due to the reduction of p53 expression together with a decrease of apoptosis in colon cancer cells.Taken together,the current findings indicate a great potential for cryptotanshinone and dihydrotanshinone against MDR colon cancer cells,in spite of P-gp overexpression and suppression of apoptosis.They are promising candidates to be developed as therapeutic agents and/or as an adjuvant therapy for colorectal cancer,especially for patients with MDR cancer types.
