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Objective:Study on the correlation between the active components of Salviea Miltiorrhizae Radix et Rhizoma screened by high-throughput sequencing and the regulation of lncRNA-mRNA in human lung adenocarcinoma A549 cells.Methods:A549 cells were cultured, and the IC 50 dose of cryptotanshinone and tanshinone ⅡA was confirmed according to the cell proliferation experiment. A549 cells were randomly divided into blank control group, cryptotanshinone group, and tanshinone IIA group using a random number table method. After 24 hours of intervention, the cell cycle was detected by flow cytometry. High-throughput sequencing technique was used to detect the expressions of lncRNA and mRNA in A549 cells in intervention group and non-intervention group. By analyzing the expression profiles of differential genes related to cryptotanshinone and tanshinone ⅡA, the obtained differential genes were analyzed by GO and KEGG. Results:The cell cycle results showed that the proportion of G0/G1 phase cells in cryptotanshinone and Tanshinone ⅡA was increased ( P<0.01), the proportion of S phase cells was decreased ( P<0.01), and the proportion of G2/M phase cells in cryptotanshinone was decreased ( P<0.01). The results of high-throughput screening showed that cryptotanshinone could up-regulate 4 698 lncRNA, down-regulate 1 557 lncRNA, up-regulate 4 810 mRNA and down-regulate 5 644 mRNA. Tanshinone ⅡA could up-regulate 1 348 lncRNA, down-regulate 1 299 lncRNA, up-regulate 4646 mRNA and down-regulate 4 741 mRNA. The function and pathway enrichment analysis of differential lncRNA and mRNA showed that the differentially expressed genes of cryptotanshinone and tanshinone ⅡA were mainly related to cell cycle, autophagy, AMPK signaling pathway, FoxO signaling pathway and EGFR signaling pathway. GAS5 may be one of the targets for the inhibitory effects of cryptotanshinone and tanshinone ⅡA. Conclusion:Cryptotanshinone and tanshinone ⅡA have certain inhibitory effects on A549 cells, and there are differentially expressed genes of lncRNA-mRNA, which are closely related to cell cycle and signal pathway.
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Pain is one of the most serious problems plaguing human health today.Drug therapy is one of the main ways to treat pain in clinic.The analgesic drugs commonly used in clinical treatment of pain are often accompanied by many side effects,the analgesic effect is still not ideal.Salvia miltiorrhiza is a traditional medici-nal material with the same origin as food and medicine.It has the functions of promoting blood circulation and removing blood stasis,relieving pain through menstrual circulation,and contains many effective ingredients such as tanshinone and salvianolic acid.Tanshinone is a kind of rosin diterpenoid compound,which mainly consists of o-quinone type and p-quinone type parent nucleus,and tanshinone Ⅱ A is the representative compound.The pharmacological mechanism of tanshinone ⅡA in labor pain mainly includes:① Regulate inflammatory factors.Inflammatory cytokines played an important role in the occurrence and progression of pain.It was found that the analgesic effect of tanshinone ⅡA was related to the anti-inflammatory effect.Tanshinone ⅡA showed anti-injuri-ous activity in various pain models,such as bone cancer pain and sciatic nerve ligation,and related studies found that tanshinone ⅡA could inhibit the expression of inflammatory factors TNF-α,IL-1β and IL-6 in the spinal cord of model rats.In the spinal nerve ligation model,tanshinone ⅡA also promoted the release of anti-inflam-matory cytokine IL-10 in the spinal cord of rats.② Regu-late signal pathways related to regulating spinal cord oxi-dation and apoptosis.Apoptosis and oxidation played an important role in the process of pain.When nerve injury was caused by stimulation,oxidative stress and apopto-sis of nerve cells were involved in the mechanism of hyperalgesia.Tanshinone ⅡA sodium sulfonate could relieve pain by regulating apoptosis-related pathways.In neuralgia model,tanshinone ⅡA could reduce the apop-tosis of spinal cord neurons by inhibiting oxidative stress response in rat spinal cord tissue.In addition,tanshinone ⅡA also decreased the expression of pro-apoptotic protein in spinal dorsal horn of CCI rats.They included caspase-3,Bcl-2,Bax protein,and enhancer binding protein homologous protein,Increased the expres-sion of anti-apoptosis protein Bcl-2.③ Inhibit the activa-tion of spinal cord glial cells.tanshinone ⅡA could exert its labor pain effect by inhibiting the activation of astro-cytes,including inhibiting the expression of chemo-therapy-induced neuralgia,inflammatory pain and inflam-matory cytokines IL-6,IL-1β and TNF-α,and inhibiting the activation of inflammatory signaling pathways related to astrocyte activation.Such as NF-κB signaling path-way,c-Jun N-terminal kinase signaling pathway,etc.In addition,tanshinone ⅡA also inhibited the activation of microglia by inhibiting the expression of CX3CR1 receptor on the surface of microglia and inhibiting the phosphoryla-tion of ERK,JNK and p38 signaling pathways.④ Decr-ease the expression of glutamate receptors in spinal cord.NMDA is an ionic glutamate receptor in the central nervous system,and its subunit NR2B is closely related to pain.The overexpression of NR2B in spinal cord could lead to the decrease of pain threshold,which was an important mechanism of pain generation.The mechani-cal threshold and thermal threshold of CCI rats were increased by tanshinone ⅡA,and the expression of spi-nal dorsal horn 2B subunit was significantly decreased after tanshinone ⅡA treatment in CCI rats.Therefore,it was concluded that the analgesic effect of tanshinone ⅡA on CCI model may be related to the decreased expres-sion of NR2B in spinal dorsal horn.In conclusion,tanshi-none ⅡA can effectively play the role of labor pain,and has great potential for development in the field of medi-cine and health products.
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Objective:To explore the mechanism of Tanshinone ⅡA(TⅡA)in improving ischemia/reperfusion(I/R)injury of H9c2 cardiomyocytes by activating Sirtuin 1(SIRT1)-adenosine 5'-monophosphateactivated protein kinase(AMPK)pathway through miR-155-5p.Methods:H9c2 cells were cultured in vitro and I/R damage model was established.After modeling,H9c2 cells were randomly divided into model group,TⅡA group,TⅡA+miR-NC group,TⅡA+miR-155-5p mimics group,10 μmol/L TⅡA was added for intervention after transfection,and the H9c2 cells supplemented with DMSO were used as control group.qRT-PCR was used to detect expression level of miR-155-5p;MTT method was used to analyze cell proliferation ability;flow cytometry was used to evaluate cell apoptosis;ELISA was used to determine the levels of TNF-α,IL-4,IL-10,IL-17,lactate dehydrogenase(LDH),malo-ndialdehyde(MDA)and superoxide dismutase(SOD);Western blot was used to detect relative expressions of SIRT1,AMPK and p-AMPK proteins.Results:Compared with control group,expression of miR-155-5p in model group was increased,cell viability was decreased,apoptosis rate and expressions of TNF-α,IL-17,LDH and MDA were increased,while expressions of IL-4,IL-10,SOD,SIRT1 and p-AMPK were decreased(P<0.05);compared with model group,expression of miR-155-5p in TⅡA group was reduced,cell viability was increased,apoptosis rate and expressions of TNF-α,IL-17,LDH and MDA were decreased,while expressions of IL-4,IL-10,SOD,SIRT1 and p-AMPK were increased(P<0.05);compared with TⅡA group and TⅡA+miR-NC group,expression of miR-155-5p in TⅡA+miR-155-5p mimics group was increased,cell viability was decreased,apoptosis rate and expressions of TNF-α,IL-17,LDH and MDA were increased,while expressions of IL-4,IL-10,SOD,SIRT1 and p-AMPK were decreased(P<0.05).Conclusion:TⅡA can improve I/R injury of H9c2 cardiomyocytes by down-regulating miR-155-5p,and its mechanism may be related to the activation of SIRT1-AMPK pathway.
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This study screened excellent carriers for co-loading tanshinone Ⅱ_A(TSA) and astragaloside Ⅳ(As) to construct antitumor nano-drug delivery systems for TSA and As. TSA-As microemulsions(TSA-As-MEs) were prepared by water titration. TSA-As metal-organic framework(MOF) nano-delivery system was prepared by loading TSA and As in MOF by the hydrothermal method. Dynamic light scattering(DLS), transmission electron microscopy(TEM), and scanning electron microscopy(SEM) were used to characterize the physicochemical properties of the two preparations. Drug loading was determined by HPLC and the effects of the two preparations on the proliferation of vascular endothelial cells, T lymphocytes, and hepatocellular carcinoma cells were detected by the CCK-8 method. The results showed that the particle size, Zeta potential, and drug loading of TSA-As-MEs were(47.69±0.71) nm,(-14.70±0.49) mV, and(0.22±0.01)%, while those of TSA-As-MOF were(258.3±25.2) nm,(-42.30 ± 1.27) mV, and 15.35%±0.01%. TSA-As-MOF was superior to TSA-As-MEs in drug loading, which could inhibit the proliferation of bEnd.3 cells at a lower concentration and improve the proliferation ability of CTLL-2 cells significantly. Therefore, MOF was preferred as an excellent carrier for TSA and As co-loading.
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Mice , Animals , Endothelial Cells , Abietanes , Cell LineРеферат
In this study, the transmittance of tanshinone Ⅱ_A(Tan Ⅱ_A) and cryptotanshinone(CTS) through the blood-prostate barrier and their distributions in the prostate tissue were compared between tanshinone extract(Tan E) treatment group and the corresponding monomer composition group under the equivalent dose conversion in vitro and in vivo. First, the human prostate epithelial cell line RWPE-1 was cultured in vitro for 21 days for the establishment of a blood-prostate barrier model, and the transmission of Tan Ⅱ_A and CTS through the barrier model was investigated after administration of Tan E and corresponding single active components. Second, SD rats were administrated with 700 mg·kg~(-1) Tan E, 29 mg·kg~(-1) CTS, and 50 mg·kg~(-1) Tan Ⅱ_A by gavage, and plasma and prostate tissue samples were collected at the time points of 2, 4, 8, 12, and 24 h. The Tan Ⅱ_A and CTS concentrations in the samples were determined. The results showed that in the cell model, the cumulative transmission amounts of CTS and Tan Ⅱ_A in the extract at each time point were higher than those of the corresponding single active components(P<0.01). In rats, after the administration of Tan E, the concentrations of Tan Ⅱ_A and CTS in rat plasma and prostate were higher than those of the corresponding single active components. This study demonstrated that the coexisting components in Tan E promoted the penetration of its main pharmacological components Tan Ⅱ_A and CTS through the blood-prostate barrier. The findings provide a theoretical and experimental basis for the application of Tan E in the clinical treatment of prostate-related diseases.
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Male , Rats , Humans , Animals , Prostate , Rats, Sprague-Dawley , Abietanes/pharmacology , PermeabilityРеферат
This study aims to investigate the molecular mechanism of tanshinone Ⅱ_(A )(TaⅡ_A) combined with endothelial progenitor cells-derived exosomes(EPCs-exos) in protecting the aortic vascular endothelial cells(AVECs) from oxidative damage via the phosphatidylinositol 3 kinase(PI3K)/protein kinase B(Akt) pathway. The AVECs induced by 1-palmitoyl-2-(5'-oxovaleroyl)-sn-glycero-3-phosphocholine(POVPC) were randomly divided into model, TaⅡ_A, EPCs-exos, and TaⅡ_A+EPCs-exos groups, and the normal cells were taken as the control group. The cell counting kit-8(CCK-8) was used to examine the cell proliferation. The lactate dehydrogenase(LDH) cytotoxicity assay kit, Matrigel assay, DCFH-DA fluorescent probe, and laser confocal microscopy were employed to examine the LDH release, tube-forming ability, cellular reactive oxygen species(ROS) level, and endothelial cell skeleton morphology, respectively. The enzyme-linked immunosorbent assay was employed to measure the expression of interleukin(IL)-1β, IL-6, and tumor necrosis factor(TNF)-α. Real-time fluorescence quantitative PCR(qRT-PCR) and Western blot were employed to determine the mRNA and protein levels, respectively, of PI3K and Akt. Compared with the control group, the model group showed decreased cell proliferation and tube-forming ability, increased LDH release, elevated ROS level, obvious cytoskeletal disruption, increased expression of IL-1β, IL-6, and TNF-α, and down-regulated mRNA and protein levels of PI3K and Akt. Compared with the model group, TaⅡ_A or EPCs-exos alone increased the cell proliferation and tube-forming ability, reduced LDH release, lowered the ROS level, repaired the damaged skeleton, decreased the expression of IL-1β, IL-6, and TNF-α, and up-regulated the mRNA and protein levels of PI3K and Akt. TaⅡ_A+EPCs-exos outperformed TaⅡ_A or EPCs-exos alone in regulating the above indexes. The results demonstrated that TaⅡ_A and EPCs-exos exerted a protective effect on POVPC-induced AVECs by activating the PI3K/Akt pathway, and the combination of the two had stronger therapeutic effect.
Тема - темы
Proto-Oncogene Proteins c-akt/metabolism , Phosphatidylinositol 3-Kinases/metabolism , Signal Transduction , Reactive Oxygen Species/metabolism , Tumor Necrosis Factor-alpha/metabolism , Interleukin-6/metabolism , Endothelium, Vascular , Oxidative Stress , Endothelial Progenitor Cells , RNA, Messenger/metabolism , AbietanesРеферат
Objective To investigate the impacts of tanshinone ⅡA(Tan ⅡA)on lipopolysaccharide(LPS)induced proliferation and apoptosis of dental pulp stem cells by regulating the fatty acid synthase(Fas)/fatty acid synthase ligand(FasL)signaling pathway.Methods Identification of human pulp stem cells(hDPSCs)isolated from the third molar of 18~20 years old patients requiring orthodontics.Lps-induced hDPSCs were treated with low,medium and high doses of Tan ⅡA,and then human recombinant FasL protein(rh FasL)was used to intervene the LPS-induced hDPSCs after high dose Tan ⅡA.Proliferation and apoptosis of hDPSCs,levels of tumor necrosis factor-α(TNF-α)and interleukin(IL)-6 in hDPSCs supernatant,proliferating cell nuclear antigen(PCNA),Cleaved aspartate-specific cysteine proteinase-3(Cleaved Caspase-3),Fas,FasL protein expression were detected.Results hDPSCs were successfully isolated.In a dose-dependent manner,Tan ⅡA promoted LPS-induced proliferation,inhibited apoptosis,up-regulated PCNA protein expression,and inhibited TNF-α,IL-6 level,Cleaved Caspase-3,Fas,and FasL protein expression.The effect of rh FasL on LPS-induced dental pulp stem cells was opposite to the above indexes(P<0.05).rh FasL attenuates the effect of high-dose Tan ⅡA on pro-liferation and apoptosis of LPS-induced dental pulp stem cells.Conclusion Tan ⅡA may promote LPS induced hDPSCs proliferation and inhibit apoptosis by inhibiting the Fas/FasL signaling pathway.
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【Objective】 To clarify the role and molecular mechanism of Tanshinone ⅡA (TanⅡA) in the pathological integration of granule cells in the dentate gyrus (DG) by using the mouse model of temporal lobe epilepsy (TLE). 【Methods】 Status epilepticus (SE) was induced in the mice with pilocarpine and treated with TanⅡA 5 mg/kg. After two months, Morris water maze was used to examine the spatial learning and memory ability and video surveillance was used to monitor spontaneous seizures. The DG was removed for staining of Timm, Prox-1, DCX and SynⅠ. PTEN, p-AKT, and p-S6 expressions were observed by Western blotting. 【Results】 TanⅡA decreased Timm score, SynⅠ, PSD-95 and pS6 levels, and increased the level of PTEN in the DG, and attenuated the formation of mossy fiber sproutings and basal dendrites of the granule cells. Video surveillance showed that TanⅡA reduced the frequency of Racine’ grade 5 seizures. 【Conclusion】 TanⅡA can effectively attenuate the abnormal integration of the granule cells in the DG by regulating PTEN/AKT/mTOR pathway and thus plays an anti-epileptic role.
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The optimal prescription of tanshinone Ⅱ_A(TSN)-glycyrrhetinic acid(GA) solid lipid nanoparticles(GT-SLNs) was explored and evaluated in vivo and in vitro, and its effect on acne after oral administration was investigated. The preparation processing and prescription were optimized and verified by single factor and response surface methodology. The in vitro release of GA and TSN in GT-SLNs was determined by ultra-performance liquid chromatography(UPLC). The effect of GT-SLNs on acne was investigated by the levels of sex hormones in mice, ear swelling model, and tissue changes in sebaceous glands, and the pharmacokinetics was evaluated. The 24-hour cumulative release rates of GA and TSN in SLNs were 65.87%±5.63% and 36.13%±2.31% respectively. After oral administration of GT-SLNs and the mixture of GA and TSN(GT-Mix), the AUC_(0-t) and AUC_(0-∞) of TSN in GT-SLNs were 1.98 times and 4.77 times those in the GT-Mix group, respectively, and the peak concentration of TSN in the GT-SLNs group was 17.2 times that in the GT-Mix group. After intragastric administration of GT-SLNs, the serum levels of testosterone(T) and the ratio of testosterone to estradiol(T/E2) in the GT-SLNs group significantly declined, and the sebaceous glands of mice were atrophied to a certain extent. The results demonstrated that obtained GT-SLNs with good encapsulation efficiency and uniform particle size could promote the release of GA and TSN. GT-SLNs displayed therapeutic efficacy on acne manifested by androgen increase, abnormal sebaceous gland secretion, and inflammatory damage.
Тема - темы
Animals , Mice , Abietanes , Acne Vulgaris/drug therapy , Drug Carriers , Glycyrrhetinic Acid , Liposomes , Nanoparticles , Particle Size , TestosteroneРеферат
ObjectiveTo predict the potential targets and possible related signaling pathways of Salviae Miltiorrhizae Radix et Rhizoma against bladder cancer (BC) based on network pharmacology and verify the potential molecular mechanism through in vitro cell experiment. MethodActive components of Salviae Miltiorrhizae Radix et Rhizoma were retrieved from Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP) and BC-related targets were searched from GeneCards and Online Mendelian Inheritance in Man (OMIM). Via Venny2.1, the potential targets of Salviae Miltiorrhizae Radix et Rhizoma against BC were screened out and the Venn diagram was plotted. Protein-protein interaction (PPI) network was constructed by STRING, followed by Gene Ontology (GO) term enrichment and Kyoto Encyclopedia of Genes and Gnomes (KEGG) pathway enrichment with DAVID. Cell Counting Kit-8 (CCK-8) assay was employed to detect the inhibitory effect of tanshinone ⅡA (Tan ⅡA), cryptotanshinone (CPT), and luteolin (LUT) at different concentration (0, 1, 2, 4, 8, 16, 32 μmol·L-1) on the proliferation of BC T24 and 5637 cells, propidium iodide (PI) staining to analyze the apoptosis of 5637 cells induced by Tan ⅡA, CPT, and LUT (0, 4, 8 μmol·L-1), and Western blotting to detect the regulatory effect of Tan ⅡA (0, 4, 8, 16 μmol·L-1) on the expression of key target proteins. ResultA total of 65 active components and 39 anti-BC targets of Salviae Miltiorrhizae Radix et Rhizoma were screened out. The anti-BC targets were mainly involved in the KEGG pathways of neuron-ligand-receptor interaction, phosphatidylinositol 3-kinases (PI3K)/protein kinase B (Akt) signaling pathway, epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor resistance, and hypoxia inducible factor (HIF)-1 signaling pathway. As for the CCK-8 assay, compared with the blank group, Tan ⅡA, CPT, and LUT significantly inhibited the proliferation of T24 and 5637 cells, particularly the 5637 cells. The half maximal inhibitory concentration (IC50) of Tan ⅡA on 5637 cells was significantly lower than that of CPT and LUT. Moreover, compared with the blank group, Tan ⅡA, CPT, and LUT all induced the apoptosis of 5637 cells, and the effect followed the order of Tan ⅡA>CPT>LUT (P<0.05). Western blot showed that Tan ⅡA significantly reduced the expression of EGFR, p-PI3K, and p-Akt in 5637 cells in a concentration-dependent manner compared with the blank group (P<0.05). ConclusionSalviae Miltiorrhizae Radix et Rhizoma exerts therapeutic effect on BC through multiple components, multiple targets, and multiple pathways. The mechanism is the likelihood that it down-regulates the expression of EGFR, p-PI3K, and p-Akt proteins, thus further inhibits cell proliferation, and induces apoptosis.
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Objective@#To investigate the inhibition of tanshinoneⅡA (TanⅡA) on the proliferation of lung cancer cells.@*Methods@#Human lung cancer cell lines A549, SK-MES-1, H446 and H460 were cultured in vitro and treated with TanⅡA at concentrations of 0.3, 0.6, 1.3, 2.5, 5.0, 10.0 mg/mL, while untreated cells served as controls. Cell proliferation was measured by using CCK-8 assay, and apoptosis was measured using flow cytometry, while the expression of apoptosis-related proteins was determined using Western blotting. The apoptotic rate of lung cancer cells was compared between Tan ⅡA-treated cells and untreated cells.@*Results@# Tan ⅡA inhibited lung cancer cell proliferation in a time-dependent and concentration-dependent manner, and the survival rates of A549, SK-MS-1, H446 and H460 cells reduced with the concentration of TanⅡA (t=4.503, 2.114, 2.103 and 3.567; all P<0.05) and the duration of TanⅡA treatment (t=5.189, 3.079, 3.023 and 3.845; all P<0.05). The 48 h half maximal inhibitory concentrations (IC50 values) of TanⅡA were (1.18±0.12), (0.78±0.08), (1.55±0.16) and (1.27±0.14) mg/mL against A549, SK-MES-1, H446 and H460 cells, respectively. Following 48 h treatment with Tan ⅡA at a concentration of 2.5 mg/mL, the apoptotic rates of A549, SK-MS-1, H446 and H460 cells were (34.97±3.78)%, (37.62±2.48)%, (18.27±2.98)% and (19.17±2.30)%, which were significantly significantly higher than those of untreated cells [(4.86±0.36) %, (3.21±0.48) %, (3.25±0.26)% and (2.66±0.19)%, all P<0.05]. Reduced Akt1 protein expression, elevated Bax/Bcl-2 expression ratio, and elevated caspase-9 and caspase-3 protein expression were detected in lung cancer cells treated with 2.5 mg/mL TanⅡA for 48 h relative to untreated cells@*Conclusion@#TanⅡA may inhibit lung cancer cell proliferation in a time- and concentration-dependent manner via the Bax/Bcl-2/caspase-9/caspase-3 pathway.
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Objective:To explore the effect of tanshinone ⅡA on myocardial remodeling in ischemia/reperfusion (I/R)-induced heart failure of rodent model.Methods:① In vivo, 30 SD rats were randomly divided into sham operation, heart failure and tanshinone ⅡA treatment group, with 10 rats in each group. The I/R model was established by ligating the left coronary artery until ST segment elevation for 30 minutes, then the ligation was removed for 2 hours as reperfusion. In the sham operation group, the rat chest was opened without artery ligation. Three days after model establishment, tanshinone ⅡA (10 mg/kg) were given intraperitoneal injected in tanshinone ⅡA group for 9 weeks. In the other two groups, normal saline was administrated in the same way. The behavioral manifestations of the rats in each group were observed; hemodynamic indexes were evaluated; Masson staining was performed to observe the degree of myocardial fibrosis; enzyme linked immunosorbent assay (ELISA) was used to detect the content of Galectin-3 in myocardial tissue; quantitative reverse transcription-polymerase chain reaction (qRT-PCR) was used to detect the expressions of collagenⅢ, collagenⅠ, matrix metalloproteinase 2 (MMP-2) and tissue inhibitor of metalloproteinase (TIMP-1). ② In vitro, rats primary cardiac fibroblasts were extracted and isolated, and divided into blank control group, angiotensinⅡ group (7-10 mmol/L angiotensinⅡ) and angiotensinⅡ+ tanshinoneⅡA group (7-10 mmol/L angiotensinⅡ+ 5-10 mmol/L tanshinone ⅡA). At 24 hours and 48 hours of culture, the cell proliferation in each group was detected by methyl thiazolyl tetrazolium (MTT); the expressions of collagenⅢ, collagenⅠ, MMP-2 and TIMP-1 were detected by qRT-PCR; the content of Galectin-3 in cardiac fibroblasts was detected by ELSIA. Results:① In vivo, the rats' activity status, hair conformity and food intake were ranked from good to bad in order of sham operation group, tanshinone ⅡA group and heart failure model group. Compared with the sham-operated group, the heart rate (HR) of the rats in the heart failure model group was significantly decreased and the heart function was significantly impaired. The mRNA and protein expression of collagenⅠ, collagenⅢ, TIMP-1 and Galectin-3 content were significantly increased, while the mRNA and protein expression of MMP-2 were significantly decreased. Compared with the heart failure model group, rats in the tanshinone ⅡA group showed significantly higher HR and improved cardiac function, significantly lower mRNA expression of collagenⅠ and collagenⅢ, significantly lower mRNA and protein expression of TIMP-1 and Galectin-3, and significantly higher mRNA and protein expression of MMP-2, and the most obvious changes were in the 9th weeks of modeling [collagenⅠ mRNA (2 -ΔΔCt): 4.70±1.19 vs. 10.21±1.62, collagenⅢ mRNA (2 -ΔΔCt): 3.03±0.46 vs. 13.84±1.93, TIMP-1 mRNA (2 -ΔΔCt): 1.90±0.19 vs. 4.55±0.43, TIMP-1/GAPDH: 0.33±0.04 vs. 0.67±0.05, Galectin-3 (ng/L): 489.93±79.30 vs. 821.72±94.09, MMP-2 mRNA (2 -ΔΔCt): 0.37±0.07 vs. 0.03±0.01, MMP-2/GAPDH: 0.69±0.09 vs. 0.21±0.04, all P < 0.05]. Masson staining showed that myocardial tissue fibrosis was obvious in the heart failure group, and the degree of fibrosis in the tanshinoneⅡA group was reduced. ② In vitro, compared with the blank control group, the proliferation rate, collagenⅠ, collagen Ⅲ and TIMP-1 expression and Galectin-3 content of myocardial fibroblasts were significantly increased, and MMP-2 expression was significantly decreased in the angiotensin group at 24 h and 48 h of culture. Compared with the angiotensin group, the proliferation rate of cardiac fibroblasts and the expression of collagenⅠ, collagen Ⅲ and TIMP-1 and the content of Galectin-3 were significantly decreased, and the expression of MMP-2 mRNA was significantly increased in the angiotensin + tanshinone ⅡA group, and the most significant changes were at 48 hours of culture [proliferation rate: (57.0±3.7)% vs. (67.0±2.4)%, collagenⅠmRNA (2 -ΔΔCt): 551.43±67.10 vs. 871.48±12.25, collagenⅢ mRNA (2 -ΔΔCt): 233.76±18.73 vs. 385.51±31.35, TIMP-1 mRNA (2 -ΔΔCt): 238.69±17.37 vs. 351.84±26.17, Galectin-3 (ng/L): 283.76±28.73 vs. 415.51±31.35, MMP-2 mRNA (2 -ΔΔCt): 108.54±12.10 vs. 51.47±6.25, all P < 0.05]. Conclusion:Tanshinone ⅡA can improve cardiac function, inhibit myocardial fibrosis and improve myocardial remodeling in rats with I/R-induced heart failure.
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OBJECTIVE:To e valuate the correlation between processing time ,color and chemical composition content in the wine-fried process of Salvia miltiorrhiza . METHODS :Wine-fried S. miltiorrhiza with different processing time (7-15 min)was prepared by yellow rice wine. The contents of salvianolic acid B ,tanshinone ⅡA and 5-HMF in raw and wine-fried S. miltiorrhiza were determined by HPLC. The colorimeter was used to determine their chromatic values [red-green axis component (a*), yellow-blue axis component (b*),lightness(L*)] and calculate the total color difference value (ΔE). Spearman ’s rho and Kendall ’s Tau-b test were adopted to validate the correlation between processing time ,chromatic value and chemical composition content. RESULTS:The contents of salvianolic acid B ,tanshinone ⅡA and 5-HMF were 17.9-70.6,2.3-3.1,0 mg/g in S. miltiorrhiza decoction pieces ;the contents of them were 14.8-68.4,1.1-3.9,0.7-34.4 mg/g in wine-fried S. miltiorrhiza . The content of salvianolic acid B at first decreased and then increased ,reaching the peak at about 9,11 min,and then gradually decreased ;the content of tanshinone ⅡA increased at first ,reached its peak about 7 min,and then gradually decreased;the content of 5-HMF increased sharply after frying 13 min. The measurement results of chromaticity values were ΔL* -5.369-2.553,Δa* -1.098-0.321, Δb* -1.471- 2.355,ΔE 0.217-5.397. Results of Spearman ’s rho and Kendall ’s Tau-b test showed that ΔE was positively correlated with processing time (the correlation coefficient were 0.517,0.389 respectively)and 5-HMF content (the correlation coefficient were 0.549,0.405 respectively)(both P<0.01). The content of tanshinone ⅡA was negatively correlated with Δb(* the correlation coefficient were -0.509,-0.391 respectively),processing time (the corr elation coefficient were -0.556,-0.420 respectively) and 5-HMF content (the correlation coefficient were -0.545,-0.392 respectively)(both P<0.01). The content of 5-HMF was positively correlated with the processing time (the correlation coefficient were 0.957,0.870 respectively)(both P<0.01). CONCLUSIONS :The contents of tanshinone ⅡA and 5-HMF in the process of wine-fried process are significantly related to the time and color. With the increase of processing time and temperature ,its color changes from red 话:0553-3844333。E-mail:liulj1@126.com yellow to yellow green ,and tends to be black in black and white;the content of tanshinone ⅡA is decreased and the content of 5-HMF is increased.
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Objective::To screen out the effective components of Salvia miltiorrhiza by establishing an in vitro model of pulmonary epithelial mesenchymal transformation. Method::Different concentrations of salvianolic acid A (10, 20, 40, 80, 160 μmol·L-1), salvianolic acid B (10, 20, 40, 80, 160 μmol·L-1), tanshinol (10, 20, 40, 80, 160 μmol·L-1), tanshinoneⅡA (10, 20, 40, 80, 160 μmol·L-1) and the blank group were applied to A549 cell, cell proliferation and cytotoxicity assay (MTS) were used to detect the proliferation effect of menthol on A549 cells.After screening the safe concentration of the active ingredients of salvia miltiorrhiza by MTS, cells were divided into blank group, model group, salvianolic acid A group, salvianolic acid B group, tanshinol group and tanshinoneⅡA.Then, the inhibitory effect of the active ingredients of salvia miltiorrhiza on the proliferation of A549 cells induced by TGF-β1 was detected by MTS. Enzyme linked immunosorbent assay (ELISA) method to detect salvia miltiorrhiza effective component of fiber protein(FN), collagen type I (COL-Ⅰ) expression. Based on the above results, the active components of salvia miltiorrhiza, which have best inhibition were screened out, and their effects on the expression of E-calcium-viscosity (E-Cad) protein were detected by Western blot. Result::Compared with blank group, salvianolic acid A 40 μmol·L-1, salvianolic acid B 160 μmol·L-1, tanshinol 160 μmol·L-1 had toxic effects on A549 cells (P<0.05). In the non-toxic concentration range, compared with the model group, salvianolic acid A 10, 20 μmol·L-1, salvianolic acid B 80 μmol·L-1 showed inhibition effect after 24 h culture (P<0.05). After 72 h culture, salvianolic acid A 5, 10, 20 μmol·L-1, salvianolic acid B 40, 80 μmol·L-1inhibition effect was very significant (P<0.01). ELISA results showed that with the blank group, model group cells the expression of FN and COL-Ⅰ increased significantly (P < 0.01). Compare with model group, salvianolic acid A 20 μmol·L-1, salvianolic acid B 80 μmol·L-1 inhibited FN and COL-Ⅰ(P<0.05). Western blot results showed that salicylic acid A and salicylic acid B had protective effects on E-Cad (P<0.01). Conclusion::Salvianolic acid A and salvianolic acid B have inhibitory effects on epithelial mesenchymal transformation by TGF-β1, which may be the main effective components of salvianolic acid in the treatment of pulmonary fibrosis.
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Objective: To investigate the protective effects of tanshinone ⅡA (tan ⅡA) against memory deficits induced by streptozotocin (STZ) in a model of sporadic Alzheimer's disease. Methods: Kunming mice were randomly divided into five groups (n=10 in each group): control group, STZ group, and tan ⅡA low, medium and high groups. Mice in the STZ and three tan ⅡA groups were given intracerebroventrically injection of STZ (3 mg/kg bilaterally, on day 1 and day 3). Tan ⅡA (20, 40, and 80 mg/kg every day) was administered intragastrically for 28 days starting from the first dose of STZ. Learning and memory were evaluated with a Morris water maze (MWM) test. Nissl method was used to observe neurons damage in the cortex and the hippocampus, and Western blot method to analyze endoplasmic reticulum stress markers and apoptosis signaling proteins. Results: Daily treatment with tan ⅡA showed a dose-dependent improvement in STZ-induced memory deficits. Nissl staining results confirmed the protective effects of tan ⅡA on cortical and hippocampal neurons damage induced by STZ. In addition, tan ⅡA markedly prevented STZ-induced abnormal expression of glucose regulated protein 78 (GRP78), initiation factor 2α (eIF2α), activating transcription factor 6 (ATF6), as well as suppressed the activation of C/EBP homologous protein (CHOP) and c-Jun N-terminal kinase (JNK) pathways. Moreover, tan ⅡA induced an up-regulation of the Bcl-2/Bax ratio and down-regulation of cleaved caspase-3 protein activity. Conclusion: Tan ⅡA prevents STZ-induced memory deficits, which may be attributed to attenuating endoplasmic reticulum stress and neuronal apoptosis.
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Objective: To evaluate the synergistic anticancer effects of the combination of apigenin (Api) and tanshinone Ⅱ A (Tan IIA), and investigate the mechanisms of pharmacological effects and their potential applications as an anticancer therapy in clinics. Methods: MTT assay were used to determine anticancer effects of the combination of Api and Tan ⅡA on BGC823, MCF7, and SMMC7721 cells. AV-PI dual stain and PI staining method were used for detecting the effect of the two drugs combination on BGC823 cell apoptosis and cell cycle. Expression of p53, BAX/BCL-2, cyclin B1 and D1 proteins were determined by Western blotting. Circular dichroism method and DNA melting point method were explored to detect interaction among the two drugs and DNA. S180 tumor xenograft mice model was used to evaluate the antitumor effects of the two drugs combination. Results: Tan IIA combined with Api exerted synergistic inhibitory effects on the proliferation of BGC823 and other tumor cells with the CI of 0.28. After tumor cell treated by combination of Tan IIA and Api, the tumor cell apoptosis was significantly enhanced and the value of BAX/BCL-2 in cells was up-regulated (P < 0.01); The levels of cyclin B1, D1 protein were changed and cell cycle arrest was increased which mainly blocked in S phase. The interaction among the two drugs and DNA was in two different ways, leading to the curves of thermal denaturation of DNA changed significantly. Furthermore, the combination of Tan IIA and Api showed a stronger inhibitory effect on tumor volume and weight in S180 mice model than monotherapy, which was similar to cyclophosphamide therapy but less side effects. Conclusion Tan IIA combined with Api exerted synergistic antitumor effects. The two drugs interacted with DNA in different ways and aggravated the cell cycle arrest, which were the key mechanisms of their synergistic antitumor effects.
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OBJECTIVE:To e valuate the clinical efficacy and economics of Sodium tanshinone Ⅱ A sulfonate injection combined with conventional medication versus conventional medication in the treatment of angina pectoris of coronary heart disease. METHODS :Using“Sodium tanshinone Ⅱ A sulfonate”“Danshentong”“angina pectoris ”as Chinese key words and “Sodium tanshinone Ⅱ A sulfonate”“Danshentong”“Angina pectoris ”as English keywords ,the studies were retrieved from Wanfang database ,CNKI,CBM,Cochrane Library ,Medline,Embase,ISI Web of Science and BIOSIS Previews during the inception to Apr. 2019. After literature screening and data extraction ,the included real world cohort studies were evaluated with bias risk tool of Cochrane systematic evaluator manual 5.2.0. Meta-analysis was conducted by using Stata 15.0 software,and publication bias of results was analyzed . The cost-effectiveness analysis was used for pharmacoeconomic evaluate ,and single factor sensitivity analysis and probability sensitivity analysis were carried out for the results of pharmacoeconomic evaluation. RESULTS : A total of 29 literatures were included ,involving 31 studies and 2 857 patients. Meta-analysis showed that clinical effective rate [RR =1.23,95%CI(1.18,1.28),P<0.001],ECG effective rate [RR =1.29,95%CI(1.20,1.39),P<0.001] and angina pectoris effective rate [RR =1.22,95%CI(1.15,1.29),P<0.001] of trial group were significantly higher than those of control group. The adverse reactions of the two groups were mild. The above results were likely to be biased in publication. Cost-effectiveness analysis showed that ICER was 72.02 of 2 groups. The sensitivity analysis showed that above results were stable. CONCLUSIONS : For patients with angina pectoris of coronary heart disease ,therapeutic efficacy of Sodium tanshinone ⅡA sulfonate injection combined with conventional medication is better that of conventional medication ,and the cost is also slightly higher. When the willingness to pay is higher than 7 202 yuan,the combination scheme has the advantage of cost-effectiveness.
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OBJECTIVE:To provide reference for safe and rational use of Sodium tanshinone ⅡA sulfonate(STS)injection in the clinic. METHODS :The information of the patients who received STS injection from Jan. 2016 to Dec. 2017 were collected from a Grade 3 hospital. According to relevant suggestions in drug package inserts ,drug utilization rationality was evaluated ,and single-factor and multi-factor analysis on the risk and influential factors for ADR/ADE were performed by group design and individual matching to examine their correlation. RESULTS :Totally 3 283 patients were included in the study. The drug use frequency were less than 1.5,and the drug utilization indexes were less than 1.0,suggesting that the hospital using STS injection was basically reasonable. Irrational use of drugs mainly included that inappropriate indications (46.48%),unreasonable solvent selection(15.84%),and excessive concentration (2.71%). Patients with renal insufficiency received STS injection ,and then the risk of ADR/ADE increased by correlation analysis (P<0.05). CONCLUSIONS :Irrational use of STS injection in clinics existed , mainly like off-label drug use ,excessive concentration ,irrational solvent selection. Drug use evaluation and monitoring should be strengthened. For patients with renal insufficiency ,it is necessary to prevent the occurrence of ADR/ADE .
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Objective To investigate the effects of fibroblast growth factor 2 (FGF-2) combined with tanshinoneⅡA on differentiation of rat bone marrow mesenchymal stem cells (BMSCs) into cardiomyocyte-like cells. Methods BMSCs were isolated and cultured. The cultured cells were identified by flow cytometry, BMSCs were divided into experimental control group, FGF-2 group, tanshinoneⅡA group and the combined induction group. The activity and value of BMSCs were detected by MTT. The Real-time PCR method was used to detect BMSCs. Expression of early myocardial transcription factors GATA-4 and Nkx2. 5; Immunocytochemical staining for detection of connexin43(Cx43)and cardiac troponin-Ⅰ(cTnI); Immunofluorescence staining for detection of desmin and Tm; The expression of desmin and tropomyosin(Tm)was detected by Western blotting. Results The cell activity and proliferation after induction were good. The expression of GATA-4 and Nkx2. 5 in the induction group were stronger than that in the experimental control group, and the difference was statistically significant (P<0. 05). The positive expression of Cx43 and cTnI in the induction group increased significantly. The markers of the combined group were the most obvious, and the difference was statistically significant (P<0. 05). The expression of Myo-specific protein desmin and Tm in the combined induction group increased significantly, and the difference was statistically significant (P<0. 05). Conclusion Both FGF-2 and tanshinoneⅡA can promote the proliferation of BMSCs and induce the differentiation of BMSCs into cardiomyocyte-like cells. The synergistic effect of the two is better than other groups.
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Objective: To observe the effects of Tanshinone ⅡA on the growth of HT29 and the expressions of TGF-β1 and Bcl-2 in colon cancer so as to provide theoretical assistance for treatment of colon cancer. Methods: Human colon cancer HT29 cells were cultured for different time after treated with various concentrations of Tanshinone ⅡA, respectively. MTT method was used to detect cell growth. The human colon cancer tumors on nude mice were randomly divided into control group and Tanshinone ⅡA treatment group, and the tumor volume was measured once a week. After 4 weeks, the tumors were preserved. The levels of TGF-β1 and Bcl-2 protein in the HT29 cells and tumor tissues were detected by Western blot. Results: With the increase of Tanshinone ⅡA concentration, the growth inhibition rate of HT29 increased; Tanshinone ⅡA downregulated the mRNA and protein levels of TGF-β1 and Bcl-2 gene in HT29. The volume inhibition rate of Tanshinone ⅡA on mouse colon cancer transplanted tumor was 28.63%. The results of Western blot showed that the levels of TGF-β1 and Bcl-2 protein were decreased in Tanshinone ⅡA group compared with control group. Conclusion: Tanshinone ⅡA has significant inhibitory effects on the growth of colon cancer cell HT29 and its xenografts model. This mechanism may be realized by the downregulation of TGF-β1 and Bcl-2 expressions via Tanshinone ⅡA.