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  • Li-ping XIANG, Li CONG, Yong ZHANG, Su-juan LIU, Xiao-lin XIE, Ping-juan BO, Xue-ping XIANG, Xiao-hua FU
    Acta Pharmaceutica Sinica. 2019, 54(2): 281-287.

    The research is aimed to investigate the effect of genistein (GEN) on the apoptosis in lipopolysaccharide (LPS)-activated RAW264.7 cells and explore the pharmacological mechanism of GEN anti-atherosclerosis (AS). RAW264.7 cells were activated by LPS, the level of TNF-α and IL-6 mRNA were detected by qRT-PCR, the expression of COX-2 and iNOS were detected by Western blot. RAW264.7 cells were pretreated with GEN for 2 h, and then incubated with LPS for 24 h. After that, CCK8 kit was used for the cell viability, Annexin V-FITC/PI kit for the apoptosis of cell. qRT-PCR was used to detect the level of CHOP, caspase-3 and miR-21. Western blot was used to detect the expression of CHOP and caspase-3. Results showed that LPS (1 000 ng·mL-1) increased the expression of TNF-α, IL-6, COX-2 and iNOS in RAW264.7 cells compared with that in control group. GEN inhibited the cell activity and the level of miR-21, promoted the expression of CHOP and caspase-3 in LPS-activated RAW264.7 cells in a dose-dependent manner. miR-21 up inhibited the expression of CHOP and caspase-3 in LPS-activated RAW264.7 cells and this process was reversed by GEN treatment. miR-21 down promoted the expression of CHOP and caspase-3, which were further enhanced by GEN. These results indicate that GEN promotes the apoptosis of RAW264.7 cells activated by LPS through down regulating miR-21 and activating endoplasmic reticulum (ER) stress pathway.

  • Ming-juan WANG, Shuai KANG, Xuan LIU, Xu WANG, Peng LIU, Zhong DAI, Shuang-cheng MA
    Acta Pharmaceutica Sinica. 2019, 54(2): 354-359.

    Though red yeast rice (RYR) has been used as medicine for centuries, few study has been reported about its biological activities related to traditional medicinal application and marketed RYR showed poor consistency in quality. In this study, with comprehensive investigation of their production processes and field acquisition samples including those from genuine producing area, an ultra performance liquid chromatographic (UPLC) method was firstly established to discriminate RYR for different applications based on their secondary metabolites fingerprint. It was performed on a CAPCELL CORE AQ column (100 mm×4.6 mm, 2.7 μm), with PDA (range:200-650 nm, extracted:237 nm) and ELSD detection. The mobile phase used was water (A) and acetonitrile (B) both containing 0.1% formic acid at gradient elution (0-15 min, 50% B→85% B (linear); 15-16 min, 85% B→50% B (linear) and maintained until 21 min), with a flow rate of 0.5 mL·min-1. The method established was fully validated in agreement with guidelines of Chinese Pharmacopeia. Common metabolites were found in RYR for same application and the fingerprints of RYR for food coloring or brewing from various manufacturers had similarities above 0.90. Meanwhile, significant differences were observed among the fingerprints for various applications and discrimination could be achieved by principal component analysis (PCA). Lovastatin was absence in RYRs for food coloring or brewing, and the fingerprint of traditional medicinal RYR was similar to that of RYR for brewing. However, standardization was required for RYR containing lovastatin because of their significant differences from various manufacturers in fingerprints and lovastatin content. The results demonstrated the feasibility to discriminate RYR for different applications by the secondary metabolites fingerprint method established in this study, which provides a scientific basis to investigate the relationship between biological activities of medicinal RYR and their corresponding secondary metabolites, and further aid their quality standardization and improvement.

  • Shi-wei ZHU, Xiao-xia GAO, Jun-sheng TIAN, Xue-mei QIN, Guan-hua DU, Yu-zhi ZHOU
    Acta Pharmaceutica Sinica. 2019, 54(2): 235-244.

    Depression, a global disease with various pathogenic factors, is seriously affecting human physical and mental health. In Chinese traditional medical theory, the disease belongs to the category of "Yu Zheng". At present, on the one hand, adverse reactions to antidepressant western medicine are becoming more and more prominent. On the other hand, the study of their antidepressant active ingredients and compatibility mechanisms has become a difficult field in the traditional Chinese medicine compounds due to their complexity. The Chinese antidepressant herb-pairs has become a hot topic in the field of antidepressant research. Herb-pair is the core of traditional Chinese medicine compound. In addition, the compound itself is a herb-pair. The study of traditional Chinese antidepressant herb-pairs is more conducive to clarify the compatibility mechanism of herb interaction and the mechanism of antidepressants on the body.Therefore, this paper systematically expounds antidepressant herb-pairs from the study of material, pharmacokinetics and efficacy, which aims at providing theoretical support for the compatibility mechanism of antidepressant herb-pairs and the research of new antidepressant drugs.

  • Rong ZHAO, Jun JIANG, Shi-chang XIAO, Chao-lin TAN, Fu-yuan WANG, Xi-ming XU, Hai-jian XIA
    Acta Pharmaceutica Sinica. 2019, 54(2): 313-320.

    To assess the therapeutic effect of Tao Hong Si Wu decoction (THSWD) on glucocorticoid-induced osteoporosis and its mechanism, the standard products of nine active components in THSWD were accurately weighed and the reserve liquid of standard product was prepared. Then a series of concentration standard liquid was obtained by dilution, and the standard curve was established. The formula of THSWD was proportionally extracted and the nine active components were determined by high performance liquid chromatography (HPLC). Naturally mated to obtain zebrafish embryos after incubation, the zebrafish larvae were cultured in blank E3 medium until the 3 days post fertilization (DPF), then divided into control group (blank E3 medium), prednisolone (PNSL) group (25 μmol·L-1 PNSL), disodium ethydronate (DE) group (15 μmol·L-1 DE and 25 μmol·L-1 PNSL), THSWD group (0.1, 1.0 or 10.0 μg·mL-1 THSWD and 25 μmol·L-1 PNSL), 15 in each group. The culture was continued until the 10 DPF. The zebrafish was anesthetized and sacrificed, stained with alizarin red S, then the bone staining was observed under a microscope. Quantitative analysis of bone mineralization area and cumulative optical density. Expression of alkaline phosphatase (AKP), type Ⅰ collagen (type Ⅰ), runt-related transcription factor 2 (RUNX-2), osteoprotegerin (OPG), tartrate-resistant acid phosphatase 5b (TRACP) and osteocalcin (OCN) were assessed by quantitative real-time PCR. The mineral contents in zebrafish were measured using inductively coupled plasma-mass spectrometry. In the THSWD water extract, the content of 9 compounds in descending order was as follows:gallic acid > hydroxysafflor yellow A > chlorogenic acid > senkyunolide H > ferulic acid > caffeic acid, senkyunolide Ⅰ > coumaric acid > benzoic acid. Compared with the control group, treatment of zebrafish with 25 μmol·L-1 prednisolone significantly reduced the area of mineralized bone and cumulative optical density, and the expression of AKP, type Ⅰ, RUNX-2, OPG, OCN gene was also significantly reduced. The levels of Ca, P, K, Mg, Zn and Fe in zebrafish were reduced by 2.8, 2.4, 13.8, 6.6, 8.0 and 8.8 times, respectively. Compared with the prednisolone group, after treating with THSWD, the area of mineralized bones and cumulative optical density value of zebrafish increased significantly and was dose-dependent, and the expression of related genes also increased. The Ca, P, K, Mg, Zn, Fe levels were significantly higher than the prednisolone group. Result show that THSWD has the potential to reverse the effect of glucocorticoid osteoporosis, the mechanism of which is to enhance the activity of osteoblast, promote the expression of bone gelatin and bone mineralization, and increase bone mass.

  • Hai-gang WANG, Ling-lei KONG, Rui WANG, Yan-xia CHEN, Shi-lun YANG, Xiao-yue ZHAO, Qi-meng ZHOU, Guan-hua DU
    Acta Pharmaceutica Sinica. 2019, 54(2): 301-307.

    This study was designed to compare the antithrombotic effects of salvianolic acid A and aspirin. The anti-platelet aggregation and anticoagulant effects of salvianolic acid A and aspirin in vitro and in vivo were investigated in normal rats. The anti-cerebral ischemia and anti-platelet aggregation effects of salvianolic acid A and aspirin were also investigated in rats with thrombotic cerebral ischemia. All animal care and experimental procedures were reviewed and approved by the Animal Ethics Committee of Chinese Academy of Medical Sciences. The results of antiplatelet aggregation in vitro and in vivo showed that salvianolic acid A could mildly inhibit adenosine diphosphate (ADP), arachidonic acid (AA) and thrombin (THR)-induced antiplatelet aggregation in a dose-dependent manner, while aspirin played a strong inhibitory effect on AA-induced platelet aggregation in vivo. The effects of salvianolic acid A and aspirin on the coagulation system were similar. At the same time, the results of maximum platelet aggregation rate (MAR) in the rat cerebral ischemia model[MARADP=(41.67±4.55)%, MARAA=(53.22±2.83)%, MARTHR=(73.33±5.04)%] indicated that salvianolic acid A could mildly inhibit ADP and AA-induced antiplatelet aggregation[MARADP=(26.13±4.60)%, MARAA=(35.53±13.73)%, P < 0.01], while aspirin played a strong inhibitory effect on AA-induced platelet aggregation[MARAA=(8.13±2.99)%]. Salvianolic acid A (10 mg·kg-1) significantly improved the neurological function, cerebral infarction volume[(10.77±7.80)%] and brain edema[(79.72±0.83)%] compared with the model group[(43.50±12.69)%, (82.25±0.89)%] (P < 0.01), while the effect of aspirin (100 mg·kg-1) was not obvious. The above results suggest that compared with aspirin, salvianolic acid A provided a mild inhibitory effect on platelet aggregation and protected against cerebral ischemia induced by thrombus. Therefore, salvianolic acid A has a good application prospect in the prevention and treatment of thrombotic diseases.

  • Xian-sheng MENG, Yong-rui BAO, Shuai WANG, Tian-jiao LI, Guo-an LUO
    Acta Pharmaceutica Sinica. 2019, 54(2): 222-227.

    The chemical composition of traditional Chinese medicine (TCM) compounds is complex, the treatment is broad, and the quality control indexes cannot accurately reflect the functional properties. According to the above problems, the authors take the research process of quality markers of Qizhiweitong granules as an example to innovate the research ideas and technical methods, and constructed five progressive steps of TCM compounds quality control and evaluation model:"based on function, to figure out the attending", "components and pharmacodynamics correlation, multiple components with multiple effects", "to analyze the components, and systematically integrate them", "spectrum and effect correlation, from a spectrum to see the efficiency", and "from the content-effect colour atla to see the quality". Based on the multiple effects of components, multiple components of multi-effect pharmacological efficacy evaluation system were established. All-time isobaric multiwavelength fusion fingerprint technology was improved and developed. "Spectrum-effect colour atla" software was research and developed for the first time, to realize the "visualization" of TCM efficacy. The aim of this work is to provide an exploratory solution for the integrated quality control of TCM compounds.

  • Xiao-jin LI, Ying-ying HUANG, Zhen YANG, Xin WU, Peng-wei ZHUANG, Yan-jun ZHANG
    Acta Pharmaceutica Sinica. 2019, 54(2): 204-210.

    The quality control of traditional Chinese medicine provides the premise of its modernization and globalization. Currently, the dual quality control based on chemical benchmark and effect benchmark has been recognized domestically and internationally. Research efforts have lead to establishment of a series of effective quality control methods based on chemical components, medicinal properties, microscopic characteristics, material constituents and pharmacodynamic targets. In the study of quality control based on chemical benchmarks, fruitful results on fingerprints, DNA barcodes, and quality markers have been achieved. However, due to a variety of factors, such as growth period, origin, growth environment and preparation process of traditional Chinese medicine, the quality control of traditional Chinese medicine based on chemical benchmarks remains difficult to fully reflect the quality of traditional Chinese medicine. At present, there is still a dispute on how to accurately reflect the quality of traditional Chinese medicines based on chemical benchmarks. For example, the index components selected in the Chinese medicine quality standards are difficult to totally reflect all the components of Chinese medicine, and the relevance between the index components versus therapeutic effect is not yet clear. In view of the complex signal network by cascade reaction and crosstalk of multi-signaling pathways within an organism, and the coordinated regulation of multi-components and multi-targets of traditional Chinese medicine, there may be different components regulating the same signal network or situations where the amount of certain chemical components within a range is not sufficient to cause a change in the signal network. Therefore, the quality control of traditional Chinese medicine based on the effect benchmark may be a useful supplement to the quality standard of traditional Chinese medicine. This paper proposes a Q-biomarker research strategy based on the effect benchmark in order to provide a methodological reference for the quality control research of traditional Chinese medicine.

  • Tie-jun ZHANG, Gang BAI, Chang-xiao LIU
    Acta Pharmaceutica Sinica. 2019, 54(2): 187-196.

    Q-marker is a new concept of quality control for traditional Chinese medicine. Since its introduction in 2016, it has received extensive support and participation from researchers and manufactures of the entire industry. In order to establishing the research level of quality markers, normative research model, we review the proposed core theory and research methods of Chinese medicine quality markers. The definition and scientific connotation of Chinese medicine quality markers are analyzed. The core theories and research methods of quality markers are summarized from five aspects:"effectiveness", "specificity", "transfer and traceability", "compatibility environment" and "measurability" of quality markers. This paper aims to provide useful theoretical and methodological guidance for studying Chinese medicine quality markers.

  • Xiao-tao HOU, Er-wei HAO, Zheng-cai DU, Zhong-shang XIA, Jia-gang DENG, Tie-jun ZHANG, Chang-xiao LIU
    Acta Pharmaceutica Sinica. 2019, 54(2): 211-221.

    The quality definition of traditional Chinese medicine (TCM) is a hot area in modern research of TCM. In recent years, the characteristics of one herb with multiple effects have been widely accepted and studied. The typical opposite-effect of herbs is considered as a special part of one herb with multiple effects, and was summarized in this paper. Sanqi was used as an example of opposite-effect herbs for developing the strategies and approaches on the Q-markers. The traditional opposite-effect should be studied by modern pharmacological research methods. The correlation of the chemical components with the opposite effects should be established in order to verify the material basis and evaluate the mechanism including targets and pathways. The unique characteristics of chemical components should be analyzed and defined. Finally, the Q-markers of the opposite effect herb will be confirmed. This paper provides a useful reference for the precise quality control of herbal opposite-effects.

  • Gang BAI, Yuan-yuan HOU, Guo-yu DING, Min JIANG, Jie GAO, Tie-jun ZHANG, Chang-xiao LIU
    Acta Pharmaceutica Sinica. 2019, 54(2): 197-203.

    The quality of traditional Chinese medicine (TCM) is the lifeline for TCM industry. The development of artificial intelligence (AI) has provided new means for the quality management of Chinese medicinal materials (CMM). In this paper, we take the quality marker (Q-marker) as a breakthrough point, focused on the research strategy from chemical markers to Q-markers, picked up the characteristics of the Q-markers from the near infrared spectrum (NIRS), and explored the feasibility of establishing the NIRS assay based on Q-marker. After integrated the biological activity detection and artificial neural network algorithm, we try to establish the relationship between the spectral properties of NIRS and specific efficacy of the CMM. Finally, the bottlenecks will be solved that related to the transmission and traceability of quality attributes in the process of TCM production, quantity change, overall quality management and so on. This system is going to improve TCM quantity scientific and intelligent supervision, and promote the upgrading of traditional TCM industry.