Mechanism of Cryptotanshinone Inhibiting Proliferation of Human Breast Cancer MCF7 Cells / 肿瘤防治研究

Objective To investigate the inhibitory effect of cryptotanshinone (CPT) on human breast cancer cell MCF7 and its mechanism. Methods The survival rate of MCF7 cells was measured by MTT assay. Cell apoptosis was detected by Annexin V/PI assay and Hoechst 33258 fluorescence staining assay. Cell cycle and reactive oxygen species were detected by flow cytometry. Cell migration and invasion were detected by cell scratch test and Transwell chamber test. The surface molecules CD44 and CD24 were detected by flow cytometry and microsphere culture. The expression of cell-associated proteins was detected by Western blot. Results CPT inhibited the proliferation of MCF7 cells in a dose-dependent manner, and the 24 h IC50 value was 19.24 μmol/L. Compared with the untreated group, the CPT-treated group showed cell cycle arrested in the S phase, and apoptosis was induced. The results of the cell scratch and Transwell chamber tests showed that CPT significantly inhibited the migration and invasion of MCF7 cells. Furthermore, CPT reduced the CD24-/LowCD44+ cell population in MCF7 cell-derived microspheres. Western blot results showed that CPT could up-regulate the expression of Bax protein, down-regulate the expression of BCL-2, PI3K-p85, Akt, N-cadherin, Twist1, Sox2, Oct4, and Nanog protein, effectively inhibit the phosphorylation of ER-α, and decrease the expression of ABCG2. Conclusion CPT can inhibit the proliferation of MCF7 cells by inhibiting the migration and invasion of MCF7 cells, decreasing the number of CD24-/lowCD44+ cells and affecting the expression of tumor stem cell-related proteins.

Progress on the antitumor activity of cryptotanshinone and its derivatives / 药学实践杂志

Cryptotanshinone is one of the effective components of traditional Chinese medicine salvia miltiorrhiza which shows good activities against a variety of tumors. Its anti-tumor effects include inhibition of tumor cell proliferation, induction of cell apoptosis, inhibition of cell migration and invasion, regulation of immune function and reversal of drug resistance. The direct anti-tumor targets include signal transducer and activator of transcription 3 (STAT3), tyrosine protein phosphatase SHP2, DNA topoisomerase 2, and other mechanisms of action include the induction of reactive oxygen species (ROS) production, regulation of estrogen and androgen receptor signaling, and inhibition of PI3K/AKT signaling pathway. In addition, many cryptotanshinone derivatives have been designed and synthesized to study the antitumor effects. The research progress of the antitumor activity of cryptotanshinone and its derivatives were reviewed in this paper to give references to the anti-tumor drug development of cryptotanshinone and its derivatives.

Research progress of cryptotanshinone on anti-fibrosis and its mechanism / 药学实践杂志

As a natural compound with high efficiency and low toxicity, cryptotanshinone (CTS) has a good anti-fibrosis effect in various organs and tissues. However, its mechanism of action has not been clearly defined, and there is no systematic literature review to describe its potential anti-fibrosis mechanism. The efficacy and mechanism of cryptotanshinone in the treatment of fibrosis in various organs were summarized and the use prospects were put forward in this paper.

Inhibitory effects and mechanisms of cryptotanshinone on the growth of tamoxifen resistant breast cancer cells / 药学学报

This study investigated the inhibitory effect and mechanisms of cryptotanshinone (CPT) on tamoxifen resistant cell MCF7-TAMR. The inhibitory effect of CPT on the viability of MCF7-TAMR cells was evaluated using the MTT assay. We found that CPT significantly inhibited the growth of MCF7-TAMR cells in a dose- and time-dependent manner. The half inhibitory concentration (IC50) is 15.14 ± 2.82 μmol·L-1 at 24 h. CPT induced cell cycle arrest of MCF7-TAMR cells at G0/G1 phase, and promoted apoptosis of MCF7-TAMR cells by upregulating intracellular levels of reactive oxygen species (ROS). Transwell results showed that CPT significantly inhibited the migration of MCF7-TAMR cells. Furthermore, CPT decreased the CD24-/lowCD44+ cell population in MCF7-TAMR cell-derived microspheres. Western blot results showed that CPT effectively inhibited the phosphorylation of estrogen receptor α (ER-α), and reduced the expression of phosphatidylinositol 3-kinase (PI3K-p85), serine-threonine protein kinase (Akt) and multidrug transporter ATP-binding cassette superfamily G member 2 (ABCG2). These results showed that CPT can induce cell apoptosis, cause cell cycle arrest, inhibit cell migration and inhibit ER-α phosphorylation, inhibit PI3K/Akt signaling pathway, reduce the number of CD24-/lowCD44+ cells and the expression of ABCG2, overcome cell drug resistance.

Construction of targeted cryptotanshinone liposomes and research on its in vitro anti-glioma effect / 药学学报

Cryptotanshinone (CPT), an active ingredient with the inhibitory effect on brain glioma cells, is trapped with poor solubility and low tumor permeability. Therefore, it is urgent to design nano drug delivery systems characterized with deep penetration and accurate targeting. In the present study, tLyp-1 modified liposomes loaded with CPT (tLipo/CPT) was prepared by emulsion solvent evaporation method. Peptide tLyp-1 which targeting tumor angiogenesis and neuropilin receptors (NRP) was modified on surface of CPT liposomes, with the aim of active targeting brain glioma cells and further release CPT precisely. The size and polymer dispersity index (PDI) of tLipo/CPT were (162.2 ± 14.6) nm and 0.24 ± 0.03. The optimal molar ratio of tLyp-1 modified on CPT liposomes was 0.5% determined by intracellular fluorescence parameters. The morphology displayed a smooth sphericity structure as determined by transmission electron microscope. Efficiency of CPT encapsulated in tLipo/CPT was detected by high performance liquid chromatography. The encapsulation efficiency of CPT was (70.06 ± 7.22) %. Liposomes modified with tLyp-1 peptide (tLipo) were internalized more than liposomes not modified with tLyp-1 (Lipo) by GL261 cells. Fluorescence intensity of tLipo in GL261 cells increased 40% than that of Lipo. Furthermore, we proved that the intake of tLipo/CPT in GL261 cells was mediated by NRP-1 receptor. MTT analysis indicated that tLipo/CPT significantly inhibit the proliferation of GL261 cells. The half maximal inhibitory concentration (IC50) was 5.70 μmol·L-1. In vitro blood-brain barrier (BBB) model experiment indicated that tLipo/CPT could penetration across BBB. Moreover, in vivo fluorescence biodistribution study indicated tail vein injection of DiR labeled tLipo after 0.5 h, DiR fluorescence could be observed in the brain of mice. Even after 24 h, DiR fluorescence still was observed in the brain. Our research certified that tLipo/CPT can penetrate the BBB and show effect of anti-glioma by inhibiting the proliferation of GL261 cells. The animal experiment was carried out in accordance with protocol evaluated and approved by the Ethics Committee of Nanjing University of Chinese Medicine.

Cryptotanshinone May Induce Ferroptosis of Human Liver Cancer HepG2 Cells / 中国医学科学院学报

Objective To observe the effect of cryptotanshinone on the ferroptosis of human liver cancer HepG2 cells. Methods The viability of the HepG2 cells cultured

Quality evaluation of Zidan Huoxue Tablets based on HPLC fingerprint pattern and chemical pattern recognition method with multi-component content analysis / 中草药

Objective: To evaluate the quality of Zidan Huoxue Tablets (ZHT) based on HPLC fingerprint and multi-component content determination combined with chemical pattern recognition method. Methods: ACE Neptune-C18 column (250 mm × 4.6 mm, 5 μm) was used with acetonitrile-0.1% phosphoric acid aqueous solution as mobile phase at a flow rate of 1.0 mL/min. The detection wavelength was 275 nm and the column temperature was 25 ℃. A total of 18 batches of ZHT was analyzed, and the quality of ZHT was evaluated by the similarity evaluation system of traditional Chinese medicine chromatographic fingerprints combined with cluster analysis and principal component analysis. Results: The fingerprint of ZHT was established. Twenty-six common peaks were identified and nine of them were identified, including 2-sodium danshensu, 8-rosmarinic acid, 9-lithospermic acid, 10-salvianolic acid B, 12-salvianolic acid A, 21-dihydrotanshinone I, 22-cryptotanshinone, 23-tanshinone I, and 25-tanshinone IIA. The similarity of fingerprints of 18 batches of ZHT was between 0.93 and 1.00. The results of cluster analysis and principal component analysis were basically consistent with similarity results. After validating the multiple component quantitative analysis condition through methodology, the average recoveries were between 98.27% and 103.42%, and the RSD were in the range of 0.86%-2.53%. The content of sodium danshensu, rosmarinic acid, lithospermic acid, salvianolic acid B, dihydrotanshinone I, cryptotanshinone, tanshinone I, tanshinone IIA in 18 batches of ZHT were in the range of 0.149%-0.218%, 0.179%-0.225%, 0.222%-0.286%, 3.570%-6.399%, 0.048%-0.136%, 0.122%-0.309%, 0.061%-0.215%, 0.093%-0.413%, respectively. Conclusion: There is a certain quality difference between different batches of ZHT. Through the combination of fingerprinting, cluster analysis and principal component analysis, the quality of ZHT can be comprehensively evaluated. The establishment of this method can provide reference for the quality control and evaluation of ZHT.

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.

Study on the therapeutic effects and mechanism of cryptotanshinone on mice with chemotherapy-induced mucositis / 药学学报

This study systematically investigated the therapeutic effects of chemotherapy-induced mucositis (CIM) by cryptotanshinone (CTS) in mice. CIM mice were prepared by intraperitoneal injection of 5-fluorouracil (5-FU) and irinotecan for 4 days. A pseudo-sterile mouse model was established by intragastric administration of mixed antibiotics (metronidazole, vancomycin, and penicillin). The body weight, disease activity index (DAI), and defecation of mice were daily monitored. The animal welfare and experimental procedures followed the rules of the Animal Ethics Committee of the Institute of Materia Medica, Chinese Academy of Medical Sciences. We determined the contents of inflammatory factors, total cholesterol (TC), triglyceride (TG), and lipase activity in serum or colonic mucosa of CIM mice. We also studied the composition and relative abundance of fecal flora. The correlation of the relative abundance of fecal microbiota and environmental factors was further analyzed. CTS significantly decreased DAI and reduced the content of interleukin 6 (IL-6), interleukin 11 (IL-11), myeloperoxidase (MPO), and diamine oxidase (DAO) in the serum of CIM mice. CTS effectively increased the content of TG while reduced TC and lipase activity in serum. Results showed the incidence of CIM in pseudoaseptic model group was significantly reduced. Meanwhile, there was no significant difference in the contents of inflammatory factors and TG/TC ratio between pseudoaseptic model group and normal control group. There was a significant difference in the diversity and composition of fecal microbiota among groups. In addition, CTS restored the composition of fecal microbiota close to normal and significantly increased the abundance of g_norank_f_Muribaculaceae. Especially, g_Ruminiclostridium and g_norank_f_Muribaculaceae exhibited a significant positive correlation to TG but a negative correlation to DAO, MPO, IL-6, lipase, and TC. Cryptotanshinone significantly increased the abundance of g_norank_f_Muribaculaceae and g_ruminococcaceae_UCG-014 in fecal microbiota of CIM mice. In conclusion, we reported CTS effectively alleviated intestinal mucositis in mice induced by 5-fluorouracil and irinotecan by regulating fecal microbiota, inflammatory factors, and serum lipid.

Cryptotanshinone Inhibits Lipid Accumulation in Differentiating 3T3-L1 Preadipocytes by Down-regulating C/EBP-α, PPAR-γ, FAS, Perilipin A, and STAT-3 / 계명의대학술지

Differentiation of preadipocyte, also named adipogenesis, leads to the phenotype of mature adipocyte that is filled with many lipid droplets. Excessive lipid accumulation in adipocytes leads to the development of obesity. In this study, we investigated the effect of 11 different natural compounds on lipid accumulation during the differentiation of 3T3-L1 preadipocytes into 3T3-L1 adipocytes. Strikingly, among the natural compounds, cryptotanshinone at 10 µM most strongly reduced triglyceride (TG) contents in 3T3-L1 cells after 8 days of the differentiation. Furthermore, cryptotanshinone at 10 µM significantly suppressed lipid accumulation in 3T3-L1 cells after 8 days of the differentiation. Cryptotanshinone at 1 to 10 µM tested did not affect the survival of 3T3-L1 cells after 8 days of the differentiation. On mechanistic levels, cryptotanshinone time-differentially decreased the expression levels of CCAAT/enhancer-binding protein-α (C/EBP-α), peroxisome proliferator-activated receptor-γ (PPAR-γ), fatty acid synthase (FAS), and perilipin A but also the phosphorylation levels of signal transducer and activator of transcription-3 (STAT-3) during the 3T3-L1 cell differentiation. Taken together, these findings demonstrate that cryptotanshinone inhibits lipid accumulation in differentiating 3T3-L1 cells, which appears to be mediated through the reduced expression and/or phosphorylation levels of C/EBP-α, PPAR-γ, FAS, Perilipin A, and STAT-3.

Effects of cryptotanshinone on airway inflammation models in asthmatic mice through tweak/fnl4 and tgf-pl/smads signaling pathways / 中国药理学通报

Aim To explore the therapeutic effect of cryptotanshinone(CTS) on airway remodeling model of asthmatic mice, and the relationship between its mechanism and the TWEAK/Fn14 and TGF-β1/Smads signaling pathway. Methods Forty female BALB/c mice were used for our study, eight mice as a group, and were assigned into five groups, namely, control group, OVA model group, CTS treatment group (20, 40 mg·kg-1), and Dex positive control group (1 mg·kg-1). HE and PAS staining were used to observe lung histopathological changes in mice; Diff-Quick staining was employed to count the types of cells; ELISA was used to detect the contents of proinflammatory cytokines in BALF; Western blot was applied to analyze the contents of TWEAK, Fn14, TGF-β1, Smad2/3, Smad4 in lung tissues; immunohistochemical method was used to detect the expression levels of TWEAK and TGF-β1 in lung tissues. Results CTS reduced the exudation of inflammatory cells and proliferation of goblet cells; CTS inhibited the generation of EOS, NEU, LYM and the total cells; CTS could reduce the level of proinflammatory cytokines of airway inflammation; the results of Western blot showed that CTS inhibited the protein expression of TWEAK, Fn14, TGF-β1, Smad2/3 and Smad4; immunohistochemical results indicated that CTS increased the expression of TWEAK and TGF-β1 in lung tissues. Conclusions CTS has therapeutic effect on the OVA-induced airway inflammation mouse model through TWEAK/Fn14 and TGF-β1/Smads signaling pathways.

Effect of cryptotanshinone on ferroptosis-related gene expression in lung cancer cells / 中国药理学通报

Aim To investigate the effects of cryptotanshinone on cell viability Aim To investigate the effects of cryptotanshinone on cell viability and ferroptosis-related gene expression in A549 and A549/DDP cells, and to explore its possible mechanisms. Methods A549 and A549/DDP cells were treated with different concentrations of CTS. Cell viability was measured by CCK-8 assay. The mRNA levels of TFR1, DMT1, IREB2, HSPB1, VDAC2, VDAC3 and GPX4 were measured by qPCR, and the protein levels of TFR1 were measured by Western blot. Results The cell viability was down-regulated by CTS in A549 and A549/DDP cells, while the cisplatin-resistant A549/DDP cells were more susceptible to CTS. After CTS treatment, the mRNA levels of ferroptosis-related genes showed different degrees of change. The mRNA levels of HSPB1 and GPX4 increased in A 5 4 9 cells , of which the mRNA levels of IREB2, VDAC2 and VDAC3 were reduced and the mRNA levels of TFR1 and DMT1 exhibited no significant change. The mRNA levels of TFR1 and IREB2 increased in A549/DDP cells, while the mRNA levels of VDAC3 decreased, and the expression levels of DMT1, HSPB1, VDAC2 and GPX4 did not changesignificantly. Conclusions Cryptotanshinone may inhibit the proliferation of lung cancer A549 and A549/DDP cells, and affect the expression of ferroptosis-re-lated genes.

Protective effect and mechanism of extracts and main components from stem and leaves of Salvia miltiorrhiza on HUVECS / 中草药

Objective: To investigate the protective effects of the extracts and active components from stems and leaves of Salvia miltiorrhiza on oxidative stress and high glucose-injured human umbilical endothelial cells (HUVECs). Methods: The models of oxidative stress and high glucose injury in HUVECs were established. The ethanol extract of S. miltiorrhiza stems and leaves (CJ), ethanol extract of S. miltiorrhiza roots (CG), water extract of S. miltiorrhiza stems and leaves (SJ), water extract of S. miltiorrhiza roots (SG), rosmarinic acid, salvianolic acid B, rutin, isoquercitrin, cryptotanshinone, aminoguanidine and VC were administrated to cells. MTT were used to observe the cell viability. The levels of glutathione peroxidase (GSH-Px), catalase (CAT), nitric oxide (NO), endothelin (ET-1), ICAM-1 and TNF-α were detected. Result:s Compared with the control group, H2O2 decreased the levels of GSH-Px, CAT and NO (P < 0.01) and increased the level of ET-1 (P < 0.01), glucose increased the levels of ICAM-1 and TNF-α (P < 0.01) and decreased the level of NO (P < 0.01). Compared with the model group, the levels of GSH-Px, CAT and NO in CJ, CG, SJ and SG groups were increased, and the levels of ET-1, ICAM-1 and TNF-α were decreased (P < 0.05, P < 0.01). VC, rosmarinic acid, salvianolic acid B and rutin high and medium dose groups, and isoquercitrin, cryptotanshinone high dose group significantly increased the levels of GSH-Px, CAT and NO (P < 0.05, P < 0.01) and decreased the level of ET-1 (P < 0.05, P < 0.01). ICAM-1 levels were significantly decreased in the high dose groups of salvianolic acid B, isoquercitrin and rutin as well as in the high and medium dose groups of rosmarinic acid (P < 0.01). In addition to the aminoguanidine group, the levels of TNF-α of other groups were significantly lower than the model group (P < 0.05, P < 0.01). Except the aminoguanidine group and isoquercetin low dose group, the levels of NO of other groups were significantly higher than the model group (P < 0.05, P < 0.01). Conclusion: In certain concentration range of alcohol extract and water extract of stems, leaves and roots of S. miltiorrhiza, rosmarinic acid, salvianolic acid B, rutin and isoquercitrin have protective effects on HUVECs injured by H2O2 and glucose. And the mechanisms are related to inhibition of intercellular adhesion molecule expression and regulation of NO and TNF-α production. This study will provide reference for the discovery and transformation of the resource value of non-medicinal stems and leaves produced during the production of S. miltiorrhiza.

Based on fingerprint and multi-index components to evaluate difference of Salvia miltiorrhiza between microwave extraction method and traditional extraction method / 中草药

Objective: To establish the UPLC fingerprint of Salvia miltiorrhiza microwave extraction method and traditional extraction method and identify the main chromatographic peaks, the chemometrics method was used to compare the chemical differences of different extraction methods. Methods: The separation was performed on a chromatographic Waters ACQUITY UPLC BEH sheid C18 column (100 mm × 2.1 mm, 1.7 μm), with acetonitrile (A)-0.1% aqueous formic acid (B) as the mobile phase gradient elution. The flow rate was 0.3 mL/min, the detection wavelength was 270 nm, and the fingerprints of traditional extraction method and microwave extraction method of S. miltiorrhiza; The content of eight index components was determined, and SPSS 22.0 software was used for principal component analysis and t-test analysis to further evaluate the difference between microwave extraction method and traditional extraction method. Results: The traditional extraction method and the microwave extraction method respectively calibrate 16 and 17 common peaks, and the content of eight index components was different. In the similarity evaluation, the fingerprints of different extraction methods of S. miltiorrhiza were compared, and the similarity was > 0.945. The content of the index components and the comprehensive mass fraction of the index components in the microwave extraction method were higher than the traditional extraction method, and the mean value of the eight index components showed that the microwave extraction method was higher than the traditional extraction method. Compared with the traditional extraction method, there were significant differences in the content of salvianic acid A sodium, protocatechuic aldehyde, rosmarinic acid, lithospermic acid, salvianolic acid B, cryptotanshinone, and tanshinone IIA (P < 0.05, 0.01). Conclusion: Through the comparison of fingerprints and chemometric methods, the overall chemical composition of S. miltiorrhiza under different extraction methods is similar. The content of chemical components is different by t-test, and the content of S. miltiorrhiza microwave extraction method is higher than traditional extraction method. It is concluded that the microwave extraction method has certain advantages compared with the traditional extraction method. The establishment of this method can lay a foundation for the wide application of microwave extraction method in the field of traditional Chinese medicine.

Chemical comparison of Salvia miltiorrhiza phloem and xylem based on fingerprint analysis and quantitative analysis of multi-components / 中草药

Objective: To comprehensively compare and evaluate the composition of Salvia miltiorrhiza phloem and xylem samples based on fingerprint analysis and quantitative analysis of multi-components. Methods: In the present study, an accurate and reliable fingerprint approach was developed using high performance liquid chromatography for chemical comparison ofphloem and xylem samples of S. miltiorrhiza. Furthermore, eight bioactive compounds including four salvianolic acids and four tanshinones in S. miltiorrhiza phloem and xylem samples were simultaneously quantified. Moreover, chemometrics methods were performed to compare and discriminate the phloem and xylem based on the quantitative data. Results: The specific fingerprints of phloem and xylem of S. miltiorrhiza were obtained, and a total of 10 common peaks were marked. The quantitative and chemometrics analysis results indicated the content of chemical components in phloem and xylem samples of S. miltiorrhiza were notably different. Obviously, the content of tanshinones were notably different between phloem and xylem samples. The content of tanshinones were significant higher in phloem compared with xylem in S. miltiorrhiza. Conclusion: The fingerprint analysis and quantitative analysis of multi-components could be a well-acceptable strategy for chemical comparison of S. miltiorrhiza phloem and xylem.

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.

Rapid Determination of Cryptotanshinone in Salvia miltiorrhiza by AOTF-NIDRS / 中国药房

OBJECTIVE:To establish rapid method for content determination of cryptotanshinone in Salvia miltiorrhiza. METHODS:The content of cryptotanshinone in sample was determined by HPLC(as reference value). AOTF-NIDRS combined with PLS was used to establish quantitative correction model for the content of cryptotanshinone in S. miltiorrhiza. According to the results of content determination of cryptotanshinone in samples,35 samples of medicinal material were collected. First-order derivative combined with smoothing filter coefficient method was used to pretreat spectrum,and optimal band range for content determination of cryptotanshinone in sample ranged 1 250-2 150 nm. RESULTS:Methodology validation of content determination of cryptotanshinone in sample was in line with the requirements. Correction mean square deviation of quantitative correction model of cryptotanshinone was 0.014 6,and predicted mean square deviation was 0.022 3,coefficient of association was 0.976 6. The internal verification deviation was 2.41% and the external verification deviation was 4.06%. CONCLUSIONS:This method is rapid,accurate,simple and pollution-free.It can be used for rapid content determination of cryptotanshinone in S.miltiorrhiza.

Research on JAK3/STAT3 mechanism of non-small cell lung cancer NCI-H1975 cells treated with cryptotanshinone com-bined with cisplatin / 实用肿瘤学杂志

Objective The aim of this study was to investigate the synergistic anti-tumor effect of cryptotanshinone and cis-platin in non-small cell lung cancer NCI-H1975 cells and its possible molecular mechanism. Methods NCI-H1975 cells were treated with control,cryptotanshinone,cisplatin or combination of cryptotanshinone and cisplatin groups(referred to as the combination group). The inhibitory rate of cell proliferation was determined in NCI-H1975 cells by CCK-8 assay;Flow cytometry was used to determine the rates of survival and cell apoptosis;The expression of apoptotic protein,anti-apoptotic protein,JAK2/STAT3 protein and its phosphorylation levels were detected in NCI-H1975 cells by Western blot. The localization and transcription activity of STAT3 cells were determined by laser confocal microscopy/luciferase assays. Results 1. The survival rates in cryptotanshinone,cisplatin and combination groups at each time-point were lower in NCI-H1975 cells than that in the control group(P<0. 05);The proliferation of NCI-H1975 cells in the combination group were inhibited when compared to the control,cryptotanshinone,or cisplatin groups(P<0. 05);2. After treatments for 24 h,the expression levels of Bcl-2 and Survivin protein were decreased(P<0. 01),the activity of Caspase3 and Caspase9 protein was increased(P<0. 01),and the expression levels of p-STAT3 and p-JAK2 protein were de-creased in NCI-H1975 cells(P<0. 01);3. After NCI-H1975 cells treated with the combination of cryptotanshinone and cisplatin, the STAT3 in the nucleus was decreased,and STAT3 activity was also decreased in the nucleus. Conclusion Cryptotanshinone com-bined with cisplatin can exert a synergistic antitumor effect on non-small cell lung cancer NCI-H1975 cells,and its mechanism is related to the inhibition of JAK2 and STAT3 phosphorylation signal pathways.

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.

Quality control research of Guanxin Danshen Capsule based on UPLC fingerprint combined with chemical pattern recognition / 中草药

Objective To establish the UPLC fingerprint of Guanxin Danshen Capsule (GDC) and conduct a systemic, comprehensive, and scientific quality evaluation of GDC using a chemical pattern recognition method. Methods Ultra-high performance liquid chromatography instrument and Acquity UPLC® HSS T3 chromatographic column was employed, the separation was performed with the mobile phase consisting of acetonitrile and 0.1% formic acid aqueous solution, and the detection wavelength was set at 256 nm to establish the UPLC fingerprint of ten batches of GDC. Then, the further quality assessment of the drug was carried out by similarity evaluation, cluster analysis (CA), principal component analysis (PCA), and orthogonal partial least squares discriminant analysis (OPLS-DA). Results In this research, 75 peaks were recognised as common peaks in the fingerprint, 13 peaks were identified using standard references, they were betaine, succinic acid, tanshinol, protocatechualdehyde, caffeic acid, rutin, galuteolin, rosmarinic acid, salvianolic acid B, salvianolic acid A, dihydrotanshinone I, cryptotanshinone, and tanshinone IIA. The similarity values of the drugs were all above 0.97, indicating a relatively stable quality of the drugs. Little difference was then discovered between the batches of the drug by CA and PCA. Finally, glycinebetaine, rutin, and salvia acid B were recognised as the quality makers using a OPLS-DA method. Conclusion The analysis method established in this study was scientific, accurate, reliable, and simple; The drug quality of GDC could be evaluated systematically and comprehensively using a fingerprint combined with chemical pattern recognition technique. Moreover, it will also lay a solid theoretical basis for the further quality control of traditional Chinese medicine and its preparations at the same time.