Preparation of two tanshinone Ⅱ_A-astragaloside Ⅳ co-loaded nano-delivery systems and in vitro antitumor activity comparison / 中国中药杂志

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.

开放科学(资源服务)标识码(OSID) Effect of Tanshinone ⅡA on abnormal integration of granule cells in the dentate gyrus of mice with chronic epilepsy / 西安交通大学学报(医学版)

【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.

Advances in pharmacological effects of tanshi-none ⅡA on pain / 中国药理学与毒理学杂志

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.

Mechanism of tanshinone ⅡA inhibiting myocardial remodeling by Galectin-3 / 中华危重病急救医学

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.

Inhibition of tanshinone ⅡA on lung cancer cell proliferation / 预防医学

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.

Molecular Mechanism of Salviae Miltiorrhizae Radix et Rhizoma against Bladder Cancer： Based on Network Pharmacology and In Vitro Experiment / 中国实验方剂学杂志

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.

Preparation of tanshinone Ⅱ_A-glycyrrhetinic acid solid lipid nanoparticles and its inhibitory effect on acne / 中国中药杂志

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.

Correlation between Processing Time ，Color and Chemical Composition Content in the Wine-fried Process of Salvia miltiorrhiza / 中国药房

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.

Synergistic antitumor effects and mechanism of apigenin and tanshinone ⅡA / 中草药

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.

Tanshinone ⅡA improves memory deficits by inhibiting the endoplasmic reticulum stress and apoptosis in Alzheimer's disease mice / 西安交通大学学报(医学版)

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.

Pharmacoeconomic Evaluation of Sodium Tanshinone ⅡA Sulfonate in the Treatment of Angina Pectoris of Co- ronary Heart Disease / 中国药房

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.

Study on Utilization Safety and Rationality of Sodium Tanshinone ⅡA Sulfonate Injection Based on “Real World ” / 中国药房

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 ．

Effect of Effective Components of Salvia Miltiorrhiza on TGF-β1 Induced Interstitial Transformation of Pulmonary Epithelial Cells / 中国实验方剂学杂志

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.

Effects of Tanshinone ⅡA on human colon cancer cell growth and expressions of TGF-β1 and Bcl-2 / 西安交通大学学报(医学版)

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.

Effect of Puerarin Combined with Tanshinone ⅡA on t-PA, PAI-1 and Oxidative Stress in DM Rats with Vascular Lesions / 中国实验方剂学杂志

Objective:To observe the protective effect and mechanism of combination of puerarin combined with tanshinone ⅡA on diabetes mellitus (DM) rats with vascular lesions. Method:The SD rats (fed with high-fat diet) were administrated with streptozotocin(STZ) through intravenous injection to make the model of diabetic vascular lesions. The successfully modeled rats were randomly divided into the model control group, the high-dose group (0.5 g·kg-1+1.0 g·kg-1), the middle-dose group (0.25 g·kg-1+0.5 g·kg-1), the low-dose group (0.05 g·kg-1+0.1 g·kg-1), the puerarin group (0.25 g·kg-1), the tanshinone ⅡA group (0.5 g·kg-1) and the positive control group (Metformin, 0.09 g·kg-1). Each group was administrated with drugs respectively by gavage for 70 days. After intervention in each group, the general conditions and body weight of the rats were observed. The contents of blood grucose and blood lipids were determined by automatic biochemical analyzer. The contents of insulin, advanced glycation end products (AGEs), superoxide dismutase(SOD), glutathione peroxidase (GSH-Px) in serum, the contents of tissue plasminogen activator (t-PA), plasminogen activator inhibitor-1 (PAI-1) and thromboxane B2 (TXB2) in plasma, as well as the contents of AGEs and oxidized low-density lipoprotein(ox-LDL) in aorta homogenate were detected by enzyme-linked immunosorbent assay(ELISA). The content of malondialdehyde(MDA) in serum was determined by chemical colorimetry. Pathological changes of coronary tissue were observed by htoxylin eosin(HE) staining. The expression of PAI-1 protein of aorta was observed by immunohistochemistry. Result:Compared with the normal control group, in the model group, the levels of blood grucose and blood lipids (PPPP2 in plasma (PPPPPPPPP2 in plasma (PPPPPPPConclusion:Puerarin combined with Tanshinone ⅡA could relieve vascular lesions of DM rats. The mechanisms may be related to the reduction of oxidative stress and the regulation of coagulation-fibrinolysis system.

Quantitative Determination of Two Flavonoids and Four Phenoquinones of Huoxue Cuyu Capsules by HPLC / 中国药房

OBJECTIVE：To establish a method for simultaneous determination of two flavonoids （rutin and kaempferol-3-O- rutinoside） and four phenoquinones （dihydrotanshinone Ⅰ，cryptotanshinone，tanshinone Ⅰ and tanshinoneⅡA） in Huoxue cuyu capsules. METHODS：HPLC method was adopted. The determination was performed on Welch Ultimate XB-C18 column with mobile phase consisted of acetonitrile-0.1％phosphoric acid solution（gradient elution）at the flow rate of 1.0 mL/min. The column temperature was set at 20 ℃，and detection wavelength was set at 270 nm. The sample size was 10 μL. RESULTS：The linear range of rutin，kaempferol-3-O-rutinoside，dihydrotanshinone Ⅰ，cryptotanshinone，tanshinone Ⅰ and tanshinoneⅡA were 172.13-860.66 μg/mL（r＝0.999 7），15.33-76.66 μg/mL（r＝0.999 8），12.81-64.06 μg/mL（r＝0.999 3），5.90-29.52 μg/mL（r＝0.999 3），5.12-25.60 μg/mL（r＝0.999 2），6.71-33.57 μg/mL（r＝0.999 7），respectively. The limits of detection were 0.08，0.01，0.01，0.01，0.01，0.01 μg/mL. The limits of quantitation were 0.27，0.02，0.03，0.03，0.03，0.03 μg/mL，respectively. RSDs of precision，stability test and repetition tests were all lower than 2.0％（n＝6）. The recoveries were 97.54％-100.25％（RSD＝1.07％，n＝6），96.90％-101.91％（RSD＝1.73％，n＝6），96.24％-102.89％（RSD＝2.32％，n＝6），97.04％-102.18％（RSD＝1.82％，n＝6），95.06％-97.73％（RSD＝1.18％，n＝6），95.59％-101.40％（RSD＝2.29％，n＝6），respectively. CONCLUSIONS：The method is sensitive，rapid，simple and has good reproducibility. It can be used for simultaneous determination of rutin， kaempferol-3-O-rutinoside， dihydrotanshinone Ⅰ， cryptotanshinone， tanshinone Ⅰ， tanshinoneⅡA in Huoxue cuyu capsules.

Mechanisms of tanshinone Ⅱ_A in reducing 4-HNE-induced hepatocyte damage by activating PPARα / 中国中药杂志

Tanshinone Ⅱ_A( Tan Ⅱ_A),the liposoluble constituents of Salvia miltiorrhiza,can not only ameliorate the lipidic metabolism and decrease the concentration of lipid peroxidation,but also resist oxidation damage,scavenge free radicals and control inflammation,with a protective effect on prognosis after liver function impairment. Therefore,the studies on the exact mechanism of Tan Ⅱ_A in protecting the liver can provide important theoretical and experimental basis for the prevention and treatment effect of Tan Ⅱ_A for liver injury. In the present study,the protective effects and mechanism of Tan Ⅱ_A on 4-hydroxynonenal( 4-HNE)-induced liver injury were investigated in vitro. Normal liver tissues NCTC 1469 cells were used to induce hepatocytes oxidative damages by 4-HNE treatment. The protective effect of Tan Ⅱ_A on hepatocytes oxidative damages was detected by release amount of lactate dehydrogenase( LDH) analysis and hoechst staining. The protein expression changes of peroxisome proliferator-activated receptor α( PPARα) and peroxisome proliferator response element( PPRE) were analyzed by Western blot analysis in NCTC 1469 cells before and after Tan Ⅱ_A treatment. The gene expression changes of fatty aldehyde dehydrogenase( FALDH) were analyzed by Real-time polymerase chain reaction( PCR) analysis. The results showed that 4-HNE increased the release amount of LDH,lowered the cell viability of NCTC 1469 cells,and Tan Ⅱ_A reversed 4-HNE-induced hepatocyte damage. Western blot analysis and RT-PCR analysis results showed that 4-HNE decreased the expression of PPARα and FALDH and increased the expression of 4-HNE. However,the expression of PPARα and FALDH were increased significantly and the expression of 4-HNE was decreased obviously after Tan Ⅱ_A treatment. This study confirmed that the curative effect of Tan Ⅱ_A was obvious on hepatocytes damage,and the mechanism may be associated with activating PPARα and FALDH expression as well as scavenging 4-HNE.

Inhibitory effect of Tanshinone Ⅱ A on the proliferation of human retinal pigment epithelial cells in hypoxia and its mechanism / 中华实验眼科杂志

Objective To investigate the effect of Tanshinone Ⅱ A on the proliferation and the signaling pathway of human retinal pigment epithelial (RPE) cells in hypoxia.Methods CoCl2(150 μ mol/L) was used to simulate hypoxic condition and the ARPE-19 cells cultured in vitro were divided into blank control group,hypoxia control group,Tanshinone Ⅱ A group and hypoxia-inducible factor-1αt (HIF-1 α) inhibitor group.The different doses of Tanshinone Ⅱ A were used to treat ARPE-19 for 24,48 and 72 hours,respectively.The inhibitory rate of cell proliferation of different groups were detected by MTT after 24,48 and 72 hours of administration cultured,and the apoptosis rate and the cell cycle distribution of cells in the hypoxia were analyzed by flow cytometry.Real-time PCR and Western blot were used to detect the expressions of mRNA and protein of HIF-1α and vascular endothelial growth factor (VEGF).Results MTT assay showed that Tanshinone Ⅱ A could inhibit the proliferation of ARPE-19 cells in a dose-and time-dependent manner,and the proliferation inhibitory rate gradually increased in the 1,5 and 10 mg/L Tanshinone Ⅱ A groups,with significant differences between any two groups (all at P＜0.05).Flow cytometry showed that the apoptosis rate of ARPE-19 in 1,5 and 10 mg/L Tanshinone Ⅱ A groups gradually increased with the elevation of Tanshinone Ⅱ A dosage,with significant differences between any two groups (all at P＜0.05).The cell proportion in the G0/G1 phase gradually increased,while the cell proportion in the S phase gradually decreased along with the elevation of Tanshinone Ⅱ A concentration,significant differences were obtained among the hypoxia control group,1,5 and 10 mg/L Tanshinone Ⅱ A groups (all at P＜0.05).RT-PCR and Western blot showed that the relative expression of VEGF mRNA,HIF-1 α and VEGF protein in the the blank control group,hypoxia control group,1,5 and 10 mg/L Tanshinone Ⅱ A groups and HIF-1α inhibitor group were significantly different (all at P＜0.05).The expression of VEGF mRNA,HIF-1α and VEGF protein decreased successively in the 1,5 and 10 mg/L Tanshinone Ⅱ A groups,with significant differences between them (all at P＜0.05).There were no significant differences between 10 mg/L Tanshinone Ⅱ A and the HIF-1 α inhibitor group of all the test indexes(all at P＞0.05).Conclusions Tanshinone Ⅱ A can inhibit the proliferation of RPE cells,induce apoptosis by arresting cells at G0/G1 phase.The mechanism is related to the suppression of HIF-1 α/VEGF signaling pathway.

Induction of tanshinone ⅡA on differentiation of human placenta-derived mesenchymal stem cells into cardiomyocytes and its mechanism / 吉林大学学报(医学版)

Objective:To investigate the induction of tanshinoneⅡA (TanⅡA) on the differentiation of human placenta-derived mesenchymal stem cells (hPDMSCs) into cardiomyocytes, and to provide an experimental basis for TanⅡA as a cardiomyocyte differentiation inducer.Methods:The hPDMSCs were treated with different concentrations of TanⅡA (0.1, 0.2, 0.4, 0.6, 0.8, 1.0, 2.0, 4.0, 6.0, 8.0, and 10.0mg·L-1) , and the nontoxic dose of TanⅡA (0.1mg·L-1) was screened by MTT assay for experiment.The hPDMSCs were divided into control group, 5-aza induction (10μmol·L-1) group, and TanⅡA induction (0.1mg·L-1) group.After culture for 20d, the expressions ofα-sarcomeric actin (α-SCA) in the cells in various groups were detected with immunohistochemistry;the positive expression rates of cardiac troponin I (cTnI) in the cells in various groups were detected with immunofluorescence, and the differentation rates of cardiomyocytes were calculated.The expression levels of GATA-binding protein 4 (GATA4) , atrial natriuretic factor (ANF) , cTnI, glycogen synthase kinase-3β (GSK-3β) andβ-catenin in the cells were detected with Western blotting method.Results:The biological characteristics of hPDMSCs accorded with the mesenchymal stem cells.The MTT results showed that when the concentration of TanⅡA was more than 0.1mg·L-1, the cell survival rates were decreased with the increase of concentration;the cells in control group showed a rapid growth trend before 12d, and the proliferation activities of the cells began to decrease on the 12th day.Compared with control group, the cell activities in 5-aza induction group and TanⅡA induction group were significantly decreased (P＜0.05) .The immunohistochemistry staining results showed that the cells in control group didn't expressα-SCA, and the cells in 5-aza induction group and TanⅡA induction group expressedα-SCA, especially in TanⅡA induction group.Compared with control group, the expression levels of GATA4 (t5-aza=2.937, P5-aza＜0.05;tTanⅡA=4.769, PTanⅡA＜0.05) , ANF (t5-aza=3.728, P5-aza＜0.05;tTanⅡA=5.912, PTanⅡA＜0.05) , cTnI (t5-aza=3.623, P5-aza＜0.05;tTanⅡA=7.153, PTanⅡA＜0.05) and GSK-3β (t5-aza=2.995, P5-aza＜0.05;tTanⅡA=5.420, PTanⅡA＜0.05) proteins in the cells in 5-aza induction group and TanⅡA induction group were significantly increased, and the expression levels ofβ-catenin (t5-aza=2.985, P5-aza＜0.05;tTanⅡA=6.951, PTanⅡA＜0.05) protein were significantly decreased;compared with 5-aza induction group, the expression levels of GATA4, ANF, and GSK-3βproteins in TanⅡA induction group were increased (P＜0.05) .Conclusion:TanⅡA can induce the differentiation of hPDMSCs into cardiomyocytes, which has better effect than 5-aza, and its mechanism may be related to inhibiting the Wnt/β-catenin signaling pathway.

Effects of Cyclodextrin Inclusion on the Degradation of Tanshinone ⅡA in Rat Intestinal Flora / 中国药学杂志

OBJECTIVE: To observe effects of β-cyclodextrin inclusion on tanshinone ⅡA in the degradation which in rat intestinal flora in vitro. METHODS: Culturing the tanshinone ⅡA and inclusion compound separately with incubation buffer of rat intestinal flora, sampling after 06121824 h. Extracting the sample then analyze content by HPLC. RESULTS: The tanshinone ⅡA is degraded obviously by rat intestinal flora in vitro and degradation speed became slower after forming the inclusion compound. After degrading 18 h, the declining proportion of tanshinone ⅡA concerntration which in tanshinone ⅡA group and mixture of tanshinone ⅡA with β-cyclodextrin group reached (74.23±2.32)% and (80.23±1.14)% while (47.45±4.01)% in β-cyclodextrin inclusion group. CONCLUSION: The tanshinone ⅡA is degraded almost completely by rat intestinal flora in vitro during 24 h, the degradation speed will be slower while forming the inclusion compound.