Preparation of tanshinone IIA self-soluble microneedles and its inhibition on proliferation of human skin fibroblasts / 中草药·英文版

OBJECTIVE@#Hypertrophic scars (HS) are a variety of skin tissue fibrosis disease that occurs in human skin, the effective therapeutic method of which is still inaccessible up to now. As a bioactive constituent of a well-known medical plant, Salvia miltiorrhiza (Danshen in Chinese), tanshinone IIA (TSA) is reported to inhibit cell proliferation in HS. Therefore, the aim of this study was to prepare TSA self-soluble microneedles to strengthen its dermal retention and break through the difficulty of significantly thickening epidermal connective tissue and stratum corneum at the HS site. The possible mechanism of action in suppressing HS was studied using human skin fibroblasts (HSF).@*METHODS@#Tanshinone IIA self-dissolving microneedles (TSA-MN) was prepared using a negative mold casting method. The prescription process of microneedle was optimized by Box-Behnken effect surface method. Different media were selected to investigate the ability of transdermal absorption and in vitro release. Furthermore, according to Cell Counting Kit-8 (CCK8) method as well as the Western blot method, the effect of TSA-MN on the biological characteristics of HSF was investigated.@*RESULTS@#With remarkable slow release effect and dermal retention, the release and transdermal properties of TSA-MN in vitro were better than both TSA and ordinary dosage forms. Its effect of HSF confirmed the essential decrease in cell motility during cell proliferation and cell migration in vitro, which plays a significant role in down-regulating the secretion of transforming growth factor-β1 (TGF-β1) in HSF and increasing the expression level of Smad7.@*CONCLUSION@#The prepared TSA self-soluble microneedles is helpful in solving the problem of hypertrophic scars, with a stable dermal retention effect after process optimization.

Effect of crosstalk between Th17 and Th9 cells on the activation of dermal vascular smooth muscle cells in systemic scleroderma and regulation of tanshinone IIA

Abstract Background: To evaluate the effect of T-helper 17 (Th17) cells and Th9 cells on the activation of dermal vascular smooth muscle cells (DVSMCs) in systemic scleroderma (SSc) and regulation of tanshinone IIA. Methods: The expression of interleukin 17 receptor (IL-17R) and interleukin 9 receptor (IL-9R) in the skin of SSc patients was assessed by immunofluorescence. The expression of IL-9 and IL-9R mRNA in peripheral blood mononuclear cells (PBMCs) of SSc patients were detected by quantitative real-time polymerase chain reaction (qRT-PCR). The proportion of Th9 cells in PBMCs of SSc patients was sorted by flow cytometry. The effect of IL-9 on the differentiation of Th17 and IL-17 on that of Th9 was detected by flow cytometry. The proportion of Th9 and Th17 cells in SSc patients was detected by flow cytometry. The level of collagen I, III, α-SMA, IL-9R, IL-17R, JNK, P38, and ERK were analyzed using western blot (WB). Results: Th9 cells were highly expressed in SSc. IL-9 stimulated the differentiation of immature T cells into Th17 cells. IL-17 induced the differentiation of immature T cells intoTh9 cells. Tanshinone IIA inhibited the differentiation of immature T lymphocytes into Th17 and Th9. WB showed that the combined action of IL-17 and IL-9 upregulated the inflammation and proliferation of DVSMCs. Anti-IL17, anti-IL9, and tanshinone IIA inhibited the functional activation of DVSMCs. Study limitations: For Th17, Th9 and vascular smooth muscle cells, the study on the signal pathway of their interaction is not thorough enough. More detailed studies are needed to explore the mechanism of cell-cell interaction. Conclusions: The current results suggested that Th17 and Th9 cells induced the activation of DVSMCs in SSc through crosstalk in vitro, and tanshinone IIA inhibited the process.

Tanshinone IIA prevents acute lung injury by regulating macrophage polarization / 中西医结合学报

OBJECTIVE@#Acute lung injury (ALI) is a serious respiratory dysfunction caused by pathogen or physical invasion. The strong induced inflammation often causes death. Tanshinone IIA (Tan-IIA) is the major constituent of Salvia miltiorrhiza Bunge and has been shown to display anti-inflammatory effects. The aim of the current study was to investigate the effects of Tan-IIA on ALI.@*METHODS@#A murine model of lipopolysaccharide (LPS)-induced ALI was used. The lungs and serum samples of mice were extracted at 3 days after treatment. ALI-induced inflammatory damages were confirmed from cytokine detections and histomorphology observations. Effects of Tan-IIA were investigated using in vivo and in vitro ALI models. Tan-IIA mechanisms were investigated by performing Western blot and flow cytometry experiments. A wound-healing assay was performed to confirm the Tan-IIA function.@*RESULTS@#The cytokine storm induced by LPS treatment was detected at 3 days after LPS treatment, and alveolar epithelial damage and lymphocyte aggregation were observed. Tan-IIA treatment attenuated the LPS-induced inflammation and reduced the levels of inflammatory cytokines released not only by inhibiting neutrophils, but also by macrophage. Moreover, we found that macrophage activation and polarization after LPS treatment were abrogated after applying the Tan-IIA treatment. An in vitro assay also confirmed that including the Tan-IIA supplement increased the relative amount of the M2 subtype and decreased that of M1. Rebalanced macrophages and Tan-IIA inhibited activations of the nuclear factor-κB and hypoxia-inducible factor pathways. Including Tan-IIA and macrophages also improved alveolar epithelial repair by regulating macrophage polarization.@*CONCLUSION@#This study found that while an LPS-induced cytokine storm exacerbated ALI, including Tan-IIA could prevent ALI-induced inflammation and improve the alveolar epithelial repair, and do so by regulating macrophage polarization.

Tanshinone IIA alleviates monocrotaline-induced pulmonary hypertension in rats through the PI3K/Akt-eNOS signaling pathway / 南方医科大学学报

OBJECTIVE@#To explore the therapeutic mechanism of tanshinone IIA in the treatment of pulmonary arterial hypertension (PAH) in rats.@*METHODS@#A total of 100 male SD rats were randomized into 5 groups (n=20), and except for those in the control group with saline injection, all the rats were injected with monocrotaline (MCT) on the back of the neck to establish models of pulmonary hypertension. Two weeks after the injection, the rat models received intraperitoneal injections of tanshinone IIA (10 mg/kg), phosphatidylinositol 3 kinase (PI3K) inhibitor (1 mg/kg), both tanshinone IIA and PI3K inhibitor, or saline (model group) on a daily basis. After 2 weeks of treatment, HE staining and α-SMA immunofluorescence staining were used to evaluate the morphology of the pulmonary vessels of the rats. The phosphorylation levels of PI3K, protein kinase B (PKB/Akt) and endothelial nitric oxide synthase (eNOS) in the lung tissue were determined with Western blotting; the levels of eNOS and NO were measured using enzyme-linked immunosorbent assay (ELISA).@*RESULTS@#The results of HE staining and α-SMA immunofluorescence staining showed that tanshinone IIA effectively inhibited MCT-induced pulmonary artery intimamedia thickening and muscularization of the pulmonary arterioles (P < 0.01). The results of Western blotting showed that treatment with tanshinone IIA significantly increased the phosphorylation levels of PI3K, Akt and eNOS proteins in the lung tissue of PAH rats; ELISA results showed that the levels of eNOS and NO were significantly decreased in the rat models after tanshinone IIA treatment (P < 0.01).@*CONCLUSION@#Treatment with tanshinone IIA can improve MCT-induced pulmonary hypertension in rats through the PI3K/Akt-eNOS signaling pathway.

Mechanism of tanshinone IIA to prevent spontaneous breast cancer in mice / 药学学报

To explore the effect of tanshinone IIA (TanIIA) on the occurrence and development of breast cancer, we employed the mouse mammary tumor virus-polyomavirus middle T antigen (MMTV-PyMT) transgenic mice as a spontaneous breast cancer mouse model. Animal welfare and experimental procedures were in accordance with the regulations of the Animal Ethics Committee of Nanjing University of Chinese Medicine. The animals were divided into control group, low-dose TanIIA treatment group (30 mg·kg-1·day-1), and high-dose TanIIA treatment group (60 mg·kg-1·day-1). The treatment was administered orally and daily for 5 weeks. The mice were sacrificed after final treatment. Mammary gland and lung were collected for histopathology studies. We evaluated the chemoprophylaxis effect of TanIIA on breast cancer in mice according to the pathological characteristics of breast cancer at different stages of development. Immunofluorescence staining were employed for blood vessel analysis. The expression levels of E-cadherin, proliferating nuclear antigen (PCNA), and oncogene c-Myc were detected by immunohistochemistry. Flow cytometry was used to analyze cell cycle and Cytoscape was used to construct drug-disease protein-protein interaction (PPI) network. Our results showed that TanIIA inhibits breast tumor progression by delaying malignancy from adenoma to early carcinoma, and inhibits blood vessel formation during tumor development. TanIIA (60 mg·kg-1·day-1) inhibits the expression levels of PCNA and c-Myc, upregulates the expression of E-cadherin. In addition, cell cycle experiments showed that the cell cycle of PyMT primary mammary cells in the high-dose TanIIA group was arrested in the G0/G1 phase. Our study demonstrated that TanIIA can significantly inhibit breast tumor progression in MMTV-PyMT mouse model, which may be related to the inhibition of angiogenic switch and cell cycle arrest.

The effects of borneol on the pharmacokinetics and brain distribution of tanshinone IIA, salvianolic acid B and ginsenoside Rg / 中国天然药物

Fufang Danshen preparation (FDP) is consisted of Salviae Miltiorrhizar Radix et Rhizoma (Danshen), Notoginseng Radix et Rhizoma (Sanqi) and Borneolum Syntheticum (borneol). FDP is usually used to treat myocardial ischemia hypoxia, cerebral ischemia and alzheimer's disease, etc. In the treatment of cerebrovascular diseases, borneol is usually used to promote the absorption and distribution of the bioactive components to proper organs, especially to the brain. The purpose of this study is investigating the effects of borneol on the pharmacokinetics and brain distribution of tanshinone IIA (TS IIA), salvianolic acid B (SAB) and ginsenoside Rg1 in FDP. Male healthy Sprague-Dawley (SD) rats were given Danshen extracts, Sanqi extracts (Panax notoginsengsaponins) or simultaneously administered Danshenextracts, Sanqi extracts and borneol. Plasma and brain samples were collected at different points in time. The concentration of TS IIA, SAB and Rg1 was determined by UPLC-MS/MS method. The main pharmacokinetics parameters of plasma and brain tissue were calculated by using Phoenix WinNolin 6.1 software. In comparison with Danshen and Sanqi alone, there were significant differences in pharmacokinetic parameters of TS IIA, SAB and Rg1, and the brain distribution of SAB and TS IIA when Danshen, Sanqi and borneol were administrated together. Borneol statistically significant shortened t

Simultaneous determination of six index components analysis in Zaoren Anshen Capsules / 中草药

Objective: To establish a method for determining the contents of schisandrin, schisandrol B, cryptotanshinone, tanshinone I and tanshinone IIA, γ-schisandrin in Zaoren Anshen Capsules (ZAC), and provide scientific method for quality control of ZAC. Methods: HPLC was used by using Thermo 120Å-C18 column, with the mobile phase consisted of acetonitrile and 0.1% acetic acid solution with gradient elution for simultaneous determination of six main index components. The detection wavelengths were set at 250 nm for schisandrin, schisandrol B, γ-schisandrin and 270 nm for cryptotanshinone, tanshinone I, and tanshinone IIA, flow rate was 1.0 mL/min, and column temperature was 30℃. Results: Schisandrin, schisandrol B, γ-schisandrin, cryptotanshinone, tanshinone I, and tanshinone IIA showed good the linear ranges relationships in the range of 4.7-3 153.6 ng (r = 0.999 9), 7.864-314.500 ng (r = 0.999 9), 14.4-1 256.9 ng (r = 0.999 9), 15.1-1 103.8 ng (r = 0.999 9), 15.3-1 532.0 ng (r = 0.999 9) and 6.134-204.500 ng (r = 0.999 9), respectively. The average recoveries were 100.4%, 98.7%, 99.4%, 100.0%, 99.3% and 100.2%, RSD were 1.4%, 2.7%, 2.2%, 2.2%, 2.5% and 2.1%, respectively. The contents of each index component in 8 batches of sample were schisandrin 1.545 7-1.909 9 mg/g, schisandrol B 0.129 8-0.235 1 mg/g, cryptotanshinone 0.508 4-0.523 4 mg/g, tanshinone I 0.111 7-0.122 3 mg/g, tanshinone IIA 0.755 8-0.874 4 mg/g, γ-schisandrin 0.120 2-0.190 1 mg/g. Conclusion: The established analytical method is highly sensitive with strong specificity and it can be used efficiently in the quality control of ZAC.

Sodium tanshinone IIA sulfonate prevents radiation-induced damage in primary rat cardiac fibroblasts / 中国天然药物

This study investigated the effects of X-ray irradiation on primary rat cardiac fibroblasts (CFs) and its potential mechanism, as well as whether sodium tanshinone IIA sulfonate (STS) has protective effect on CFs and its possible mechanism. Our data demonstrated that X-rays inhibited cell growth and increased oxidative stress in CFs, and STS mitigated X-ray-induced injury. Enzyme-linked immuno-sorbent assay showed that X-rays increased the levels of secreted angiotensin II (Ang II) and brain natriuretic peptide (BNP). STS inhibited the X-ray-induced increases in Ang II and BNP release. Apoptosis and cell cycle of CFs were analyzed using flow cytometry. X-rays induced apoptosis in CFs, whereas STS inhibited apoptosis in CFs after X-ray irradiation. X-rays induced S-phase cell cycle arrest in CFs, which could be reversed by STS. X-rays increased the expression of phosphorylated-P38/P38, cleaved caspase-3 and caspase-3 as well as decreased the expression of phosphorylated extracellular signal-regulated kinase 1/2 (ERK 1/2)/ERK 1/2 and B cell lymphoma 2 (Bcl-2)/Bcl-2 associated X protein (BAX) in CFs, as shown by Western blotting. STS mitigated the X-ray radiation-induced expression changes of these proteins. In conclusion, our results demonstrated that STS may potentially be developed as a medical countermeasure to mitigate radiation-induced cardiac damage.

Simultaneous determination of seven index components in Naoxintong capsule by HPLC wavelength switching method / 中国药科大学学报

@#An HPLC-DAD wavelength switching method(240 nm, 280 nm, 316 nm, 403 nm)was developed for simultaneous determination of seven index components: hydroxysafflor yellow A, paeoniflorin, ferulic acid, salvianolic acid B, kaempferol, formononetin and tanshinone IIA in Naoxintong capsule. The qualities of different batches of Naoxintong capsules were evaluated by statistical analysis. Seven index components in 20 batches of Naoxintong capsules were simultaneously determined by HPLC wavelength switching method with Capcell PAK C18 MG II column(250 mm × 4. 6 mm, 5. 0 μm). The mobile phase consisted of methanol-acetonitrile(25 ∶75, A)-0. 1% formic acid aqueous solution(B)with a gradient elution program and a flow rate of 1. 0 mL/min, and the column temperature was 30 °C. The results were analyzed by statistical analysis to evaluate the differences in the quality of Naoxintong capsules. Results showed that the seven active components were well separated and showed good linearity hydroxysafflor yellow A(403 nm)2. 30- 11. 50 mg/L(r=0. 999 2), paeoniflorin(240 nm)8. 81- 44. 05 mg/L(r=0. 999 6), ferulic acid(316 nm)1. 22- 6. 10 mg/L(r=0. 999 6), salvianolic acid B(280 nm)11. 61- 58. 05 mg/L(r=0. 999 4), kaempferol(403 nm)1. 16-5. 80 mg/L(r=0. 999 4), formononetin(240 nm)0. 12- 0. 60 mg/L(r=0. 999 5)and tanshinone IIA(280 nm)2. 28- 11. 40 mg/L(r=0. 999 5). The precision was good and RSD was less than 2. 0%, The repeatability was good and RSD was less than 2. 0%. The stability was good in 24 h. The average recoveries were between 97. 35%- 101. 02% and RSD was less than 2. 0%. The contents of target components in Naoxintong capsules, hydroxysafflor yellow A was 0. 213- 0. 369 mg/g, paeoniflorin was 1. 535- 3. 217 mg/g, ferulic acid was 0. 153- 0. 236 mg/g, salvianolic acid B was 2. 563- 3. 271 mg/g, kaempferol was 0. 103- 0. 181 mg/g, formononetin was 0. 022- 0. 028 mg/g, and tanshinone IIA was 0. 466- 0. 698 mg/g. HPLC wavelength change and gradient elution method was established for simultaneous determination of seven index components in Naoxintong capsule. The method is accurate, sensitive, reliable, and repeatable, and can be used for the quality control of Naoxintong capsule.

Molecular biological mechanism of tanshinone IIA in treatment of coronary heart disease based on network pharmacology and bioinformatics / 中草药

Objective To analyze the molecular biological mechanism of tanshinone IIA in the treatment of coronary heart disease (CHD) based on network pharmacology and bioinformatics methods. Methods The targets of tanshinone IIA were screened by uploading the chemical structure to PharmMapper database. Related targets of CHD were screened by OMIM, GeneCards, and CTD databases. The above data were imported into STRING database for PPI network analysis. Protein interaction network was constructed using Cytoscape. Gene Ontology analysis and enrichment analysis of KEGG signaling pathway were performed by Cluego. Systemsdock database was used for system molecular docking, and iGEMDOCK software was used for molecular docking to test the binding of tanshinone IIA to the targets of coronary heart disease. Results A total of 173 possible potential targets of tanshinone IIA, 42 targets related to CHD and 49 signal pathways were identified. Conclusion Tanshinone IIA has the characteristics of multi-target and multi-pathway in the treatment of CHD, and its mechanism may be through the regulation of blood pressure, cell metabolism, angiogenesis, endocrine, reactive oxygen metabolism, and other bioprocesses during the development of CHD.

Effect of tanshinone IIA on chemosensitivity of doxorubicin in breast cancer cells and its related mechanisms / 中草药

Objective To assess the effect of tanshinone IIA on the chemosensitivity of doxorubicin in breast cancer cells and investigate its related mechanisms. Methods MCF-7 and MCF-7/dox cells were respectively treated with tanshinone IIA, doxorubicin, and doxorubicin combined with tanshinone IIA. MTS assay was used to detect cell proliferation; Flow cytometry was used to analyze cell apoptosis; Scratch assay was used to evaluate cell migration; Western blotting was used to detect the expressions of APC, β-catenin, E-cadherin, MMP-2, and MMP-9. Results Tanshinone IIA could significantly enhance the inhibitory effects of doxorubicin on cell proliferation and migration of MCF-7 and MCF-7/dox cells and the effect of doxorubicin on inducing cell apoptosis. Compared to MCF-7 cells, the protein expression of APC in MCF-7/dox cells was significantly decreased (P < 0.05) while the expression of β-catenin in MCF-7/dox cells was significantly increased (P < 0.05). Compared to treatment with doxorubicin alone, combined treatment of doxorubicin and tanshinone IIA could significantly up-regulate the protein expression of APC and E-cadherin (P < 0.05) and down-regulate the protein expression of β-catenin, MMP-2, and MMP-9 (P < 0.05). Conclusion The APC/β-catenin pathway was involved in the development of doxorubicin resistance in breast cancer cells. Tanshinone IIA enhanced the chemosensitivity of doxorubicin in breast cancer cells through regulating APC/β-catenin pathway.

Simultaneous determination of seven constituents in Compound Yi’anning Pills by HPLC coupled with wavelength switching method / 中草药

Objective: To establish an HPLC method for the simultaneous content determination of seven constituents in Compound Yi’anning Pills (CYP). Methods: The determination was performed on Shim-pack GIST C18 column with mobile phase consisted of acetonitrile-0.1% phosphoric acid (gradient elution: 0-5 min, 5% acetonitrile; 5-12 min, 5%-12% acetonitrile; 12-17 min, 12%-20% acetonitrile; 17-30 min, 20% acetonitrile; 30-36 min, 20%-50% acetonitrile; 36-45 min, 50%-80% acetonitrile; 45-70 min, 80% acetonitrile) at the flow rate of 1.0 mL/min. The detection wavelength was set at 203 nm for notoginsenoside R1, ginsenoside Rg1, and ginsenoside Rb1, 220 nm for schisantherrin A, 250 nm for schisandrin, 268 nm for salvianolic acid B, and 270 nm for tanshinone IIA. The column temperature was set at 35 ℃ and the injection volume was 10 μL. Results: The linear range of detection concentration was 0.015-0.150 mg/mL for notoginsenoside R1, 0.077-0.766 mg/mL for schisandrin, 0.006-0.062 mg/mL for salvianolic acid B, 0.009-0.092 mg/mL for buddleodide, 0.050-0.503 mg/mL for ginsenoside Rg1, 0.067-0.670 mg/mL for ginsenoside Rb1, and 0.011-0.108 mg/mL for tanshinone IIA. Average recoveries respectively were 99.5%, 99.8%, 99.2%, 98.9%, 96.9%, 98.8%, and 97.1%. Conclusion: The method is simple, effective, and accurate, which can be used for simultaneous determination of seven active constituents in CYP.

Optimization of spray drying technology of Liqi Huoxue Compound extract by Box-Behnken response surface method combined with G1-entropy weight method / 中草药

Objective: To optimize the spray drying process of Liqi Huoxue Compound (LHC) extract and evaluate the quality of extract powder. Methods: The drug concentration, inlet air temperature and air flow were used as independent variables, and the comprehensive scores of ferulic acid, tanshinone IIA, salvianolic acid B, puerarin content and flour extraction rate were used as the response values, and the test data were binomial fitting. The mathematical relationship between the comprehensive scoring index and their respective variables was established, and Box-Behnken design-response surface method combined with G1-entropy weight method was used to optimize the spray drying process of LHC extract. Results: The optimal spray drying process was a specific solution gravity of 1.08, inlet air temperature of 140 ℃, and an air flow of 40 m3/h. Under such conditions, the average content of ferulic acid, tanshinone IIA, salvianolic acid B and puerarin by the spray drying was 0.300 4 mg/g, 0.338 0 mg/g, 18.526 0 mg/g and 7.508 8 mg/g, respectively. The flour yield was 81.49%, the overall score was 94.69, and the RSD value was 0.99%. Conclusion: The preferred spray drying process of LHC extract is stable and feasible.

Lower clearance of sodium tanshinone IIA sulfonate in coronary heart disease patients and the effect of total bilirubin: a population pharmacokinetics analysis / 中国天然药物

This study developed a population pharmacokinetic model for sodium tanshinone IIA sulfonate (STS) in healthy volunteers and coronary heart disease (CHD) patients in order to identify significant covariates for the pharmacokinetics of STS. Blood samples were obtained by intense sampling approach from 10 healthy volunteers and sparse sampling from 25 CHD patients, and a population pharmacokinetic analysis was performed by nonlinear mixed-effect modeling. The final model was evaluated by bootstrap and visual predictive check. A total of 230 plasma concentrations were included, 137 from healthy volunteers and 93 from CHD patients. It was a two-compartment model with first-order elimination. The typical value of the apparent clearance (CL) of STS in CHD patients with total bilirubin (TBIL) level of 10 μmol(L was 48.7 L(h with inter individual variability of 27.4%, whereas that in healthy volunteers with the same TBIL level was 63.1 L(h. Residual variability was described by a proportional error model and estimated at 5.2%. The CL of STS in CHD patients was lower than that in healthy volunteers and decreased when TBIL levels increased. The bootstrap and visual predictive check confirmed the stability and validity of the final model. These results suggested that STS dosage adjustment might be considered based on TBIL levels in CHD patients.

Synthesis and biological evaluation of novel tanshinone IIA derivatives for treating pain / 中国天然药物

Due to ineffectiveness and side effects of existing analgesics, chronic pain has become one of the most complex and difficult problems in the clinic. Monoacylglycerol lipase (MAGL) is an essential hydrolase in the endocannabinoid system and has been identified as a potential target for the treatment of pain. In the present study, we designed and synthesized twelve tanshinone IIA analogs and screened their activity against MAGL. Selected compounds were tested for analgesic activity in vivo, with the acetic acid writhing test model. Among the test compounds, compound III-3 (IC 120 nmol·L) showed significant activity against MAGL and ameliorated the clinical progression in the mouse pain model. Additionally, compound III-3, substitution with N-methyl-2-morpholinoacetamide, demonstrated improved solubility relative to tanshinone IIA.

Synthesis and biological evaluation of novel tanshinone IIA derivatives for treating pain / 中国天然药物

Due to ineffectiveness and side effects of existing analgesics, chronic pain has become one of the most complex and difficult problems in the clinic. Monoacylglycerol lipase (MAGL) is an essential hydrolase in the endocannabinoid system and has been identified as a potential target for the treatment of pain. In the present study, we designed and synthesized twelve tanshinone IIA analogs and screened their activity against MAGL. Selected compounds were tested for analgesic activity in vivo, with the acetic acid writhing test model. Among the test compounds, compound III-3 (IC 120 nmol·L) showed significant activity against MAGL and ameliorated the clinical progression in the mouse pain model. Additionally, compound III-3, substitution with N-methyl-2-morpholinoacetamide, demonstrated improved solubility relative to tanshinone IIA.

Simultaneous Determination of Six Components in Ganyan Kangfu Pills by HPLC / 中国药师

Objective:To establish a wavelength switching HPLC with gradient elution for the simultaneous determination of six active components, [(R,S)-goitrin, salvianolic acid B, tanshinone IIA, luteolin-7-glucoside, 4-hydroxyacetophenone and scoparone ] in Ganyan Kangfu pills .Methods:An Agilent TC-C18 (250 mm ×4.6 mm, 5 μm) chromatographic column was used with the mobile phase consisting of acetonitrile and 0.1％ phosphoric acid solution with gradient elution .The flow rate was 0.9 ml· min-1 , and the column temperature was 25 ℃.( R,S)-Goitrin was detected at 245 nm, salvianolic acid B and tanshinone IIA were detected at 270 nm, luteolin-7-glucoside was detected at 348 nm, 4-hydroxyacetophenone and scoparone were detected at 278 nm.The sample volume was 10 μl.Results:The linear range of (R,S)-goitrin, salvianolic acid B, tanshinone ⅡA, luteolin-7-glucoside, 4-hydroxyacetophe-none and scoparone was 1.99-49.75 μg· ml-1(r=0.9999), 18.66-466.50 μg· ml-1(r=0.9994), 2.25-56.25 μg· ml-1(r=0.9998), 2.62-65.50 μg· ml-1(r=0.9998), 2.48-62.00 μg· ml-1(r=0.9992) and 2.55-63.75μg· ml-1(r=0.9996), respectively.The average recovery was 98.42％(RSD=0.83％), 99.56％(RSD=1.04％), 97.96％(RSD=1.53％), 96.84％(RSD=0.78％), 98.10％(RSD=1.44％) and 97.82％(RSD=1.34％)(n=6), respectively.Conclusion: The method with good reproducibility is simple and accurate , which provides a new way for the quality evaluation of Ganyan Kangfu pills .

Metformin and tanshinone llA as ethnodrugs protect heart against injury by inhibiting microRNA-1 / 中国药理学与毒理学杂志

OBJECTIVE To explore the protection mechanism of metformin and tanshinone IIA on myocardial injury. METHODS The cultured neonatal rat ventricular cells (NRVCs) were exposed to 100 μmol·L-1H2O2to simulate the in vitro model of ischemia-reperfusion injury.MTT,TUNEL and Viability/Cytotoxicity Assay were used to evaluate the effect of metformin on the viability of cardiomyocytes after treated with H2O2. The target of miR-1 was verified by Dual luciferase reporter assay. ChIP analyses was adopted to reveal the relationship between C/EBP β and miR-1.Tanshinone IIA was administrated daily for 7 d before ligation of the left anterior descending artery (LAD) and lasted for 3 months after LAD.Whole-cell patch-clamp techniques were used to measure the inward rectifying K+current(IK1)in rat isolated ventricular myocytes.GRP94,p-AMPKα,C/EBP β,CHOP,Caspase-3,Kir2.1,p38 MAPK, Cx43, MEF2 and SRF levels were analyzed by Western blot and miR-1 level was quantified by Real-time PCR.RESULTS The expression of miR-1 was significantly increased in NRVCs exposed to H2O2 in vitro. miR-1 was shown to target the 3′-untranslated region (UTR) of GRP94, which results in the accumulation of un/misfolded proteins,leading to the endoplasmic reticulum(ER)stress.C/EBP β directly induces the upregulation of miR-1 by binding to its promoter.Furthermore,metformin,a direct allosteric AMPK activator, significantly reduces C/EBP β and miR-1 levels comparing with control group. Similarly, tanshinone IIA decreased the incidence of arrhythmias and relieved ischemia-induced injury.Moreover, tanshinone IIA depressed the elevated miR-1 level and inhibited the activation of p38 MAPK and heart special transcription factors SRF and MEF2 in ischemic cardiomyocytes. CONCLUSION Metformin protects cardiomyocytes against H2O2damage through AMPK/C/EBPβ/miR-1/GRP94 pathway.Tanshi-none IIA play a role in protection cardiomyocytes from ischemic injury based on inhibiting miR-1 expres-sion through p38 MAPK signal pathway.

Tanshinone IIA inhibits apoptosis, invasion and migration of esophageal cancer EC9706 and KYSE70 cells by regulating epithelial-mesenchymal transition / 中国肿瘤生物治疗杂志

@# Objective: To investigate the effect of tanshinone IIA on the invasion and migration of esophageal cancer EC9706 and KYSE70 cells, and to explore the underlying mechanism. Methods: Esophageal cancer cells (EC9706 and KYSE70) were divided into 4 groups: control group, tanshinone IIA groups (2, 4, 6 μg/ml). Cell prliferation viability was measured by CCK-8; Apoptosis was detected by flow cytometry; Invasion was tested by Transwell assay; And migration was measured by Scratch assay. The mRNA and protein levels of E-cadherin, Snail-2, Vimentin and N-cadherin were tested by quantitative Real-time reverse transcription PCR (qRT-PCR) and Western blotting, respectively. Results: Tanshinone IIAat concentrations less than 6 μg/ml did not affect the cell viability of esophageal cancer EC9706 and KYSE70 cells. The apoptosis in tanshinone IIA (4, 6 μg/ml) groups was significantly higher than that in control group ( P < 0.01). The number of invasive cells per field and wound-healing rate in tanshinone IIA(2, 4, 6 μg/ml) groups were significantly lower than those in control group (all P <0.01). Moreover, the cell morphology was transformed from a spindle-shaped mesenchymal form into epithelial morphology after tanshinone IIA treatment. Compared with control group, the expression of E-cadherin in tanshinone IIA groups (2, 4, 6 μg/ml) was significantly up-regulated while the expressions of Snail-2, Vimentin and N-cadherin were significantly down-regulated (all P <0.01). Conclusion: Tanshinone IIApromotes apoptosis and attenuates the invasion and migration of esophageal cancer cells by inhibiting the epithelial-mesenchymal transition.

Modeling extraction process of Salvia miltiorrhiza based on multi-source information fusion technology / 中草药

Objective: The potency of multi-source information fusion technology was explored to improve the model calibration and prediction performance of Chinese medicine extraction process. Methods: The ethanol extraction process for isolating fat-soluble components from Salvia miltiorrhiza was taken as the research carrier. S. miltiorrhiza from different sources were collected to simulate the fluctuation of materials. The changes of process parameters were simulated by design of experimental (DOE), and the process near infrared spectra (NIRS) were used as the process state variables. The contents of tanshinone IIA, cryptotanshinone, and tanshinone I were determined by HPLC. The raw material properties, process parameters and process state variables were combined as independent variables. The content of effective components in the extract was taken as the dependent variable. The partial least squares (PLS) algorithm was used to establish the quality prediction model of the extracts. Results: The modeling results respectively showed that the RMSECV was 0.172 8 mg/g, RMSEP was 0.031 7 mg/g, RPD was 6.91 (tanshinone IIA); RMSECV was 0.153 4 mg/g, RMSEP was 0.024 2 mg/g, RPD was 4.02 (cryptotanshinone); RMSECV was 0.117 1 mg/g, RMSEP was 0.043 2 mg/g, RPD was 4.76 (tanshinone I). Conclusion: The calibration and prediction performance of multi-source information fusion model are better than the conventional model, which can effectively improve the quality predictability and controllability of S. miltiorrhiza extract.