Volume 10,Issue 2,2018 Table of Contents

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  • 1  Scientific understanding of toxicity and safety of Chinese medicines
    Chang-xiao Liu
    2018, 10(2):107-107.
    [Abstract](498) [HTML](0) [PDF 0.00 Byte](11)
    Abstract:
    Scientific understanding of toxicity and safety of Chinese medicines Traditional Chinese medicine is a treasure of traditional culture and medicine which should be inherited and developed. How to correctly understand the toxicity of Chinese materia medica (CMM) is an important guarantee for safe drug use. The history of the application and research of the toxicity of CMM has been long, and the Chinese medicine community has never stopped exploring the issues related to the safety of CMM. As early as in the ancient time, the meaning of poison and medicine was the same, and poisons were commonly used as the term for "drug bait". During the pre-Qin period, a book on medicine and doctor “Zhou-Li: Tian-Guan Zai-Zhai” had recorded the words of "a physician's order of a doctor's prescription and a poison medicine". Although the words of the ancients may not be entirely correct, it reflects an unignorable aspect of the medicine world. With the development of society and science, the understanding of Chinese medicine has been continuously deepened, and the concept has gradually changed. The safety issue has also been paid attention to by the world's medical and pharmaceutical circles. We know that toxicity is a serious adverse effect of drugs on the body, which reflects the safety of a drug performance. Drug toxicity and adverse reactions can cause visceral tissue damage and dysfunction, and cause the body to undergo pathological changes or even death. The concept of complete toxicity of modern Chinese medicines should also include acute and sub-acute toxicity, chronic toxicity, and special toxicity such as carcinogenesis, mutagenesis, gene-toxicity, and addiction. Based on the above toxicological studies, in-depth toxicological mechanisms and health hazards can be beneficial to enhance the understanding of traditional medicines. Some research results published in domestic and foreign scientific journals are sufficient to illustrate the contribution of Chinese and foreign scientists to traditional medicine research. This special issue has published four articles on the toxicity and safety of Chinese medicine. From one aspect, we see the achievements of authors in this field. In a topic on "Application and prospect of toxicity quality markers (Q-markers) of Chinese materia medica based on metabolomics" (CHM 2018, 10(2): 108-116), authors reviewed that the toxicity Q-markers of CMM were explained from the following four aspects: 1) The establishment of toxicity prediction model of CMM based on metabolomics, recognition, and relative techniques; 2) The discovery of toxicity Q-markers of CMM based on metabolomic and toxicity mechanism research; 3) The research on the formation of toxicity Q-markers based on metabolomics from biosynthetic pathway and the secondary metabolites analysis of toxicity Q-markers; and 4) The susceptibility study of toxicity Q-markers based on metabolomics, key targets, and metabolic pathways of toxicity Q-markers to explain the target organ toxicity. Therefore, authors suggested that the application of metabolomics helped to systematically and comprehensively discover the quality control indicators related to the CMM safety, and it can provide rational evidences for the clinical toxicity of CMM. We know that the concept of Q-marker is possible to monitor the quality of Chinese Herbal medicines (CHMs) in the whole process, and control final products. However, as the complexity of CHM components and the great varieties of the extrinsic harmful residues, such as mycotoxins, pesticides, heavy metals, and sulfites, it is difficult to ensure the quality of CHMs. In the review paper "Extrinsic harmful residues in Chinese herbal medicines: types, detection and safety evaluation" (CHM 2018, 10(2): 117-136), authors summarized the potential harm, limit standards, detection methods of the harmful residues, and provided reference for the formulation or amendment of relative regulations to promote the healthy and stable development of CHMs, the information is value to establish a complete system with Q-marker evaluating the safety and quality of CHMs. In order to determine the toxic potential of Realgar-Indigo naturalis formula (RIF), Prof. Gao’s team performed a research titled "Synergistic effect and different toxicities of adjuvant components of Realgar-Indigo naturalis Formula" (CHM 2018, 10(2): 137-144). The assays acute toxicity and sub chronic toxicity were to evaluate the toxic potential of Realgar and the adjuvant components of RIF in addition to Realgar's synergy with these adjuvant components. Based on these experimental results presented, researchers found that the RIF toxicity was the result of the soluble arsenic in Realgar, and Indigo naturalis, and Salvia miltiorrhiza in the RIF exert completely opposite effects on the toxicity. Therefore, the intelligent explanation is not only a possible for the compatibility of the formula, but also the RIF study may be viewed as a classic case of CMM research on compatibility. The research paper of Prof. Zhang’s team titled “Toxicity-efficacy converting of ginseng combined with Fuzi Banxia incompatibility in heart failure stage of Cor pulmonale” (CHM 2018, 10(2): 145-151) used a heart failure model to evaluate the toxicity-efficacy and explore mechanism of ginseng combined with Fuzi Banxia. The study result exhibited that ginseng compatible environment could attenuate cardiac toxicity of Fuzi Banxia incompatibility intervening in the heart failure stage of Cor pulmonale, and improve cardiac function, which may be related to the expression of hypertrophy and apoptosis associated genes in the occurrence and development of heart failure. From the limited information and research results in these four articles, it can be understood the some safety issues of CMM from different perspectives, it can be seen that compatibility will affect their safety and effectiveness, and also affect the quality of medicines. Also, the effects are far more complex than those of single ingredient. Although the modern biology technology, such as genomics, proteomics and metabolomics technology, has been widely used in the study of mechanism of drug action, its mechanism research far fail to be elucidated and has a long way to go due to the complex components in CMM.
    2  Application and Prospect of Toxicity Quality Markers of Traditional Chinese Medicine based on Metabolomics
    Peng-jie Zhang a Ya-mei Li a Ya-ni Zhang a Wei Huang a Yu-bo Li a Yan-jun Zhang a Chang-xiao Liu b
    2018, 10(2):108-115. DOI: 10.1016/j.chmed.2018.02.001
    [Abstract](606) [HTML](0) [PDF 0.00 Byte](14)
    Abstract:
    In view of the problem that the quality evaluation indicators divorced from effectiveness and safety in Chinese materia medica (CMM), Prof. Chang-xiao Liu et al have put forward a concept of quality markers (Q-markers) of CMM, which provided a new thought for improving the CMM quality control method. The toxicity Q-markers of CMM are described as follows, the early toxicity prediction of CMM, the discovery, formation and susceptibility research of toxicity Q-markers of CMM. Metabolomics is a systematic biology approach, closely related to the toxicity phenotype of organism, has unique advantages in toxicity Q-markers research. The toxicity Q-markers of CMM are explained from the following four aspects in this review: 1) the establishment of toxicity prediction model of CMM, which is based on metabolomics, pattern recognition, and other techniques; 2) the discovery of toxicity Q-markers of CMM based on metabolomic, which is meaning that the toxicity Q-markers are interpreted regarding to the chemical composition analysis of CMM, preliminary screening and verification of the toxicity Q-markers and toxicity mechanism research; 3) the research on the formation of toxicity Q-markers based on metabolomics, which is described from biosynthetic pathway analysis and the secondary metabolites analysis of toxicity Q-markers; and 4) the susceptibility study of toxicity Q-markers based on metabolomics, namely, the key targets, and metabolic pathways of toxicity Q-markers are explained according to the target organ toxicity susceptibility caused by CMM, and then the toxicity susceptibility mechanism of CMM is clarified. Therefore, the application of metabolomics helps to systematically and comprehensively discover the quality control indicators related to the CMM safety, which further provides a rational evidence for the clinical application of toxicity of CMM.
    3  Extrinsic harmful residues in Chinese herbal medicines: types, detection and safety evaluation
    Cong-min Liu Jia-an Qin Xiao-wen Dou Mei-hua Yang Xiao-bo Sun
    2018, 10(2):116-135. DOI: 10.1016/j.chmed.2018.02.002
    [Abstract](751) [HTML](0) [PDF 0.00 Byte](16)
    Abstract:
    Chinese herbal medicines (CHMs) are playing important roles in the treatment of diseases and human health care throughout the world. However the extrinsic harmful residues have been threatening the therapies by destroying CHMs qualtiy which hinders the international trades of CHMs. This review summarized the extrinsic harmful residues contaminating CHMs in types, detection methods, national and international regulations, and made safety evaluations according to their contaminated levels, aiming to establish understanding among nations or organizations and provide references for the formulation or amendment of relative regulations.
    4  Synergistic Effect and Different Toxicities of Adjuvant Components of Realgar-Indigo naturalis Formula
    Huan-Hua Xu a b Zeng-Chun Ma a b Qiao-Li Shic Shi-Han Yangb La Jiangb Xiang-Mei Chend Yue Gao a b
    2018, 10(2):136-143. DOI: 10.1016/j.chmed.2018.03.001
    [Abstract](558) [HTML](0) [PDF 0.00 Byte](9)
    Abstract:
    Objective Realgar-Indigo naturalis formula (RIF) is a well-known arsenic-containingpreparationthat is used to treat acute promyelocytic leukemia (APL) in China. In recent multicenter clinical trials, complete remission rates in APL patients have ranged from 96.08% to 100%. RIF has a satisfactory therapeutic effect, but its safety isa widespread concern, since the preparation contains arsenic, a wide-rangingand naturally occurring toxicant. In this study, in order to determine the toxic potential of RIF, acute toxicity and subchronic toxicity assayswere performed to evaluate the toxic potential of realgar and the adjuvant components of RIFin addition to realgar’s synergy with theseadjuvant components. Methods KM mice and Wistar rats were selected forthese experiments. To evaluate acute toxicity, the toxic effects of a single dose ofa gradient of concentrations of realgarwere firstly determined. Then, the toxic effects of combinations of gradient doses of realgar and fixed doses of I. naturalis and S. miltiorrhizawere evaluated.Results The results showed that when realgar was used alone, the LD50 was 2756.73 mg/kg (equivalent to 23.6mg/kg As2O3).However, theLD50dropped to 936.90 mg/kg when realgar was used with I. naturalis.By contrast, the LD50increased to 7538.86 mg/kg when realgar was used with S. miltiorrhiza. Hence, I. naturalis strengthened the toxicity of realgar, whereas S. miltiorrhizadisplayed the opposite effect. The subchronic toxicity assessment results revealed a trend that was consistent with acute toxicity.Changes in the levels of different valence states of arsenic were also taken into account.The test results of the effects of in vitro combinations of realgar and adjuvant components on soluble arsenic dissolution showed that I. naturalis increased the level of soluble arsenic in realgar extractsand I. naturalis suspensions when the realgar/I. naturalis ratio was 2, 1.5, and 1.0.However, S. miltiorrhizadid not affect it. Conclusion Based on the collective experimental results presented here, it can be concluded that the toxicity of RIF is the result of the soluble arsenic in realgar and that the I. naturalis and S. miltiorrhiza in the RIF exert completely opposite effects on the toxicity of realgar. This maybe an intelligent explanation for the compatibility of this formula, and this RIF study may therefore be viewed as a classic case of traditional Chinese medicine research on compatibility.
    5  Toxicity-efficacy converting of ginseng combined with Fuzi Banxia incompatibility in heart failure stage of Cor pulmonale
    Ying-ying Huang Xiao-jin Li Kai-xin Liu Peng-wei Zhuang Yan-jun Zhang
    2018, 10(2):144-150. DOI: 10.1016/j.chmed.2018.03.010
    [Abstract](533) [HTML](0) [PDF 0.00 Byte](10)
    Abstract:
    Objective Fuzi Banxia is one of eighteen antagonisms, previous studies have shown that the incompatibility could play special effects in the specific condition of diseases and appropriate compatible environment. The present study aims to evaluate the toxicity-efficacy of ginseng combined with Fuzi Banxia incompatibility intervening in the heart failure stage of cor pulmonale and to explore its mechanism. Methods Monocrotaline (MCT)-induced cor pulmonale were used in this study. Ultra high-resolution small animal ultrasound real-time imaging system and the right heart catheterization were used to estimate cardiac function. Semi automatic biochemical analyzer was used to test myocardial enzyme LDH, CK, and CK-MB in serum. The heart tissues were stained with HE, and TUNEL assay was used to assess the pathomorphological changes and myocardial apoptosis. The expression of hypertrophy and apoptosis associated genes: ANP, BNP, β-MHC, Bax, and Bcl-2 in the right ventricle were determined by RT-PCR. Results Fuzi Banxia combined with ginseng obviously attenuated mortality, decreased RVHI, and increased cardiac index; RVSP and mPAP were significantly reduced, and EF and FS were raised obviously; Myocardial enzymes LDH, CK, and CK-MB were pronounced attenuated; heart diameter reduced, right ventricular dilatation was significantly decreased, inflammatory cell infiltration notably reduced, and cardiac apoptosis rate was decreased obviously. Meanwhile the expression of hypertrophy-related ANP, BNP, and β-MHC mRNA were up-regulated, the expression of apoptosis-related Bax mRNA was down-regulated, and the expression of anti-apoptosis-related Bcl-2 mRNA and Bcl-2/Bax ratio were up-regulated. Conclusion Ginseng compatible environment could attenuate cardiac toxicity of Fuzi Banxia incompatibility intervening in the heart failure stage of cor pulmonale, and improve cardiac function, which may be related to the expression of hypertrophy and apoptosis associated genes, and thus delay the occurrence and development of heart failure.
    6  Larvicidal Potential of Selected Indigenous Lichens against Three Mosquito Species- Culex quinquefasciatus, Aedes aegypti, and Anopheles stephensi
    Syed Zameer Ahmed Khader Sidhra Syed Zameer Ahmed Kisore Perundurai Venkatesh Kamaraj Chinnaperumal Sanjeeva Nayaka
    2018, 10(2):151-155. DOI: 10.1016/j.chmed.2018.03.002
    [Abstract](478) [HTML](0) [PDF 0.00 Byte](10)
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    Objective Mosquitoes are the major transmitting vectors of serious human diseases, causing millions of deaths every year with undesirable effects, including toxicity to non-target organisms. Some plants with insecticidal properties have been used in recent years for the control of a variety of pest insects and vectors. In the quest for alternative natural biological control agents against mosquito larvae lichens were selected. Method Larvicidal activity was assessed with methanolic extracts of Parmotrema reticulatum, Parmotrema kamatti, Parmotrema tinctorum, Parmelia erumpens, Leptogium papilosum,and Roccella montagnei against Aedes aegypti, Anopheles stephensi, and Culex quinquefasciatus. The standard WHO protocols with minor modifications were adopted and the bioassay was evaluated at the concentrations of 100-500 μg/mL for each lichen. Since all the lichen extracts showed complete mortality against C. quinquefasciatus in 100 μg/mL, the concentrations were decreased to 100, 50, 25, 12.5, and 6.25 μg/mL for C. quinquefasciatus. Larval mortality was observed for 24 h after treatment. Results All the lichen extracts exhibited activity against third instar larvae of A. aegypti and A. stephensi at 100 μg/mL, and 100% mortality was observed against the vector C. quinquefasciatus at 100 μg/mL. The highest larvicidal activity was found with L. papilosum against A. aegypti (LC50 = 81.127 μg/mL) and A. stephensi (LC50=89.10 μg/mL). Similarly, P. tinctorum and R. montagnei when tested against C. quinquefasciatus with minimum concentration <100 μg/mL exhibited significant activity with LC50 valuesof 5.32 and 6.97 μg/mL. Conclusion The bioassay results revealed larvicidal potential of lichens especially against C. quinquefasciatus with high mortality even at lower concertation. Hence, lichens can be used as an ideal sustainable approach for the control of lymphatic filariasis caused by vector C. quinquefasciatus.
    7  Inhibition of metabolic disorders in vivo and in vitro by main constituent of Coreopsis tinctoria
    Bao-ping Jiang Qiu-yue Lv Jia-mei Xiang Liang Le Ke-ping Hu Li-jia Xu Pei-gen Xiao
    2018, 10(2):156-167. DOI: 10.1016/j.chmed.2018.03.004
    [Abstract](483) [HTML](0) [PDF 0.00 Byte](29)
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    Objective To investigate the effects of the ethyl acetate extract of Coreopsis tinctoria (EAEC) on insulin resistance (IR) in rats fed a high-fat diet. Methods Male Sprague-Dawley (SD) rats were fed a HFD (60% fat) supplemented with EAEC for 8 weeks. The administration of EAEC to the rats with HFD-induced insulin resistance reduced hyperglycemia, plasma levels of insulin, and steatosis in the liver. Metabolomic study was used to analyse the metabolic levels of the high glucose-treated cells, control cells and marein-treated cells. Results High glucose and high fat conditions caused a significant increase in blood glucose, insulin, serum TC, TG and LDL-C levels, leading to abnormal IR in rats. However, treatment with EAEC protects against HFD-induced IR by improving the fasting serum glucose homeostasis and lipid homeostasis. The high glucose conditions significantly decreased glycogen synthesis and increased PEPCK, G6Pase and Krebs cycle-related enzyme protein levels, leading to an abnormal metabolic state in HepG2 cells. However, treatment with marein improved IR by increasing glucose uptake and glycogen synthesis and by downregulating PEPCK and G6Pase protein levels. The statistical analysis of the HPLC/MS data demonstrated that marein could restore the normal metabolic state. Conclusion The results revealed that EAEC ameliorates IR in rats, and marein has the potential to improve IR by ameliorating glucose metabolism disorders.
    8  Salidroside shows a particular pharmacokinetic property in the model rats of myocardial ischemia
    Hui-lin Gu Run-bin Sun Fei Fei Li-xiang A Hao-xue Gao Ming-xue Tao Si-qi Feng Na Yang Yue Zhang Ji-ye Aa Guang-ji Wang
    2018, 10(2):168-175. DOI: 10.1016/j.chmed.2018.03.009
    [Abstract](909) [HTML](0) [PDF 0.00 Byte](8)
    Abstract:
    Objective Salidroside showed potential pharmacological effect on plateau hypoxia and cardiovascular disease like myocardial ischemia. However, pharmacokinetic differences have not been assessed between the pathological model and the normal animals. This study focused on evaluating the pharmacokinetic properties of salidroside in animals with myocardial ischemia. Methods A reproducible and sensitive method was established and optimized based on liquid chromatography tandem mass spectrometry (LC–MS/MS) to determine salidroside in rats plasma. The data showed the AUC0–∞ and Cmax of salidroside proportionally increased along with dose elevation after singly intragastric administration of salidroside at a dose of 20, 50, and 100 mg/kg. Results Compared to the single dose, the Cmax, and AUC0–8h of salidroside markedly decreased while CL/F and V/F increased after multiple dosing. However, the Cmax and AUC0–8h of ischemic model rats were 0.35 and 0.39 fold lower than those in normal rats after a single dose at 50 mg/kg, with an increased CL/F and V/F. Surprisingly, after a consecutive administration of salidroside for 7 d, the mean Cmax, AUC0–8h increased 2.89 and 2.61 fold higher than a single dose in model rats, and even 2.28 and 4.03 fold higher than the normal controls after multiple doses. All the above fold values were statistically different (P < 0.01). Conclusion The particular PK properties of salidroside in ischemic model rats were presented in our study for the first time, suggesting that myocardial ischemia greatly affected pharmacokinetics exposure of the orally administrated salidroside after a single or multiple doses.
    9  Identification and quality analysis of Panax notoginseng and Panax vietnamensis var. fuscidicus through integrated DNA barcoding and HPLC
    Juan Yang Lin-lin Dong Guang-fei Wei Hao-yu Hu Guang-wei Zhu Jie- Zhang Shi-lin Chen
    2018, 10(2):176-182. DOI: 10.1016/j.chmed.2018.03.008
    [Abstract](1085) [HTML](0) [PDF 0.00 Byte](10)
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    Objective Root or rhizome of Panax notoginseng (Sanqi) is known for its eutherapeutic effects. Panax vietnamensis var. fuscidicus, called Yesanqi or Yuenan sanqi by local residents, is also commercially available. They are similar in morphology, leading to serious safety problems in clinical medication. It is necessary to find the rapid and efficient methods to identify them. Methods P. notoginseng and P. vietnamensis var. fuscidicus were identified by DNA barcoding based on the ITS2 sequence. Notoginsenoside R1 and ginsenosides (Rb1, Rg1, Re, Rd, Rc, and Rb2) were analyzed in the roots, fibrils, stems, leaves, and flowers of P. notoginseng and P. vietnamensis var. fuscidicus using high-performance liquid chromatography (HPLC). Results P. notoginseng and P. vietnamensis var. fuscidicus were separated into branches of divergent clusters, and P. vietnamensis var. fuscidicus and Panax vietnamensis were clustered into a clade with 98% similarity according to DNA barcoding analysis. The chemical compositions of P. notoginseng and P. vietnamensis var. fuscidicus were similar in roots; while their compositions and contents of the notoginsenoside R1 and ginsenosides in flowers, leaves, stems, and fibrils were different. Conclusion ITS2 is a rapid and efficient method to identify P. notoginseng and P. vietnamensis var. fuscidicus. HPLC analysis indicated that pharmacological action might be different between P. notoginseng and P. vietnamensis var. fuscidicus.
    10  Molecular Mechanism of Quercitrin on Osteogenic Differentiation and Adipogenic Differentiation of Rat Bone Marrow Stromal Stem Cells(rBMSCs)
    Zi-yi Guan Lan-ying Chen Xue-liang Li Ya-ru Cui Rong-hua Liu
    2018, 10(2):183-189. DOI: 10.1016/j.chmed.2018.01.009
    [Abstract](982) [HTML](0) [PDF 0.00 Byte](12)
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    Objective The study was designed to investigate the molecular mechanism of quercitrin on osteogenic differentiation and adipogenic differentiation of rBMSCs. Methods rBMSCs were harvested from SD rats, and determination of alkaline phosphatase (ALP) activity, quantification of mineralization by Alizarin Red S staining, and the mRNA expression of osteogenic differentiation markers (Runx2, BMP-2, and OSX) by RT-PCR after rBMSCs stimulated by osteogenic induction with (0.1-10) μg/mL of quercitrin, quantification of Lipid droplet by Oil Red O staining and the mRNA expression of adipogenic differentiation marker (PPARγ, C/EBPα, and aP2) by RT-PCR after rBMSCs stimulated by adipogenic induction with (0.1-10) μg/mL of quercitrin. Results Quercitrin can up-regulate the mRNA expression of osteogenic differentiation markers (Runx2, BMP-2, and OSX) and increase ALP activity and mineralization after osteogenic induction, on the other hand quercitrin can suppress the mRNA expression of adipogenic differentiation markers (PPARγ, C/EBPα, and aP2) and decrease lipid droplet after adipogenic induction. Conclusion This study suggested that quercitrin not only stimulated osteogenic differentiation but also inhibited adipogenic differentiation of rBMSCs, which was associated with the up-regulation of Runx2, BMP-2, and OSX mRNA expression and the down-regulation of PPARγ, C/EBPα, and aP2 mRNA expression.
    11  Protection of Shuxuetong combined with aspirin against cerebral ischemia/reperfusion injury through ameliorating coagulation and fibrinolysis system
    Yong-mei Wu Ya-dong Zhai Yu-ting Zhou Shi-min Tang Xin-yue Li Xiang-bao Meng Hui Zhang Gui-bo Sun Xiao-bo Sun
    2018, 10(2):190-197. DOI: 10.1016/j.chmed.2018.03.011
    [Abstract](861) [HTML](0) [PDF 0.00 Byte](12)
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    Objective To explore the influence of the combination of Shuxuetong (SXT) and aspirin on coagulation and fibrinolytic system of rats. Methods Suture method was applied to establish focal cerebral ischemia-reperfusion injury models in rats. A total of 50 SD rats were randomly divided into sham group, middle cerebral artery occlusion/reperfusion (MCAO/R) group, aspirin (10 mg/kg) group, SXT (0.6 mL/kg) group, and SXT (0.6 mg/kg) + aspirin (10 mg/kg) group. The neurological deficits were assessed according to Longa's grade 5 scoring method. The cerebral edema was detected by measuring the content of water in brain tissue. The volume of cerebral infarction was observed by 2,3,5-Triphenyltetrazolium chloride (TTC) staining. Blood plasma was collected by abdominal aortic method to test maximum platelet aggregation rate and four blood coagulation. CD61, CD62p, 6-keto prostaglandin F1α (6-keto-PGF1α), antithrombin III (AT-III), D-dimer, plasminogen activator inhibitor-1 (PAI-1), tissue factor (TF), tissue plasminogen activator (t-PA), platelet thromboxane B2 (TXB2), and von Willebrand factor (vWF) content in rat plasma were detected by ELISA. Results SXT combined with aspirin could improve the neurological deficits, alleviate cerebral edema, and decrease the cerebral infarct value. Compared with the sham operation group, fibrinogen (FIB), 6-PGF1α, AT-III, and t-PA in model group were significantly decreased; Compared with the model group, the above-mentioned indexes in SXT and aspirin treatment group were significantly increased. The prothrombin time (PT), activated partial thromboplastin time (APTT), thrombin time (TT), D-dimer, PAI-1, TF, TXB2, and vWF of the model group were significantly increased; the above-mentioned indexes in blood SXT + aspirin treated group were significantly decreased. There was a significant difference between the combined group and SXT group. The maximum concentration of plateletsin aspirin treated rats was significantly decreased, however, MPAR was reversed in SXT + aspirin treated group. Conclusion SXT combined with aspirin can effectively inhibit platelet activation, regulate the maximum concentration of platelets, and improve coagulation function and fibrinolysis system.
    12  Effect of high exposure of chlorogenic acid on lipid accumulation and oxidative stress in oleic acid-treated HepG2 cells
    Ya-yun Liu Ting Zhai Qing-qing Yu Jing Zhu Yong Chen
    2018, 10(2):198-204. DOI: 10.1016/j.chmed.2018.03.005
    [Abstract](578) [HTML](0) [PDF 0.00 Byte](11)
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    Objective To evaluate the effect of high concentration of chlorogenic acid (CGA) on non-alcoholic fatty liver disease (NAFLD) in normal and oleic acid (OA) treated HepG2 cells, as well as the underlying mechanism involved in the fat accumulation, oxidative stress and insulin resistance (IR) induced by CGA treatment. Methods OA (0.5 mmol/L) induced hepatic steatosis was established in HepG2 cells as an in vitro model of NAFLD. The normal and OA-treated HepG2 cells were treated by CGA (0, 0.5, 1, and 2 mmol/L) for 24 h, then cellular lipid droplets, reactive oxygen species (ROS), and glucose uptake were evaluated by Oil Red O staining and cellular biochemical assays, respectively. Signaling pathways involved in?adipogenesis including SREBP-1c and PNPLA3, oxidative stress, and IR including CYP2E1 and CYP4A, were investigated by Western blot and RT-qPCR. Results CGA (0.5, 1, and 2 mmol/L) treatment increased the cellular lipid droplets and the expression of SREBP-1c and PNPLA3 in the tested cells. Additionally, 2-NBDG uptake was significantly increased, whereas the cellular ROS and protein levels of CYP2E1 and CYP4A were significantly decreased in OA-treated cells. Conclusion Our results suggest that high concentrations of CGA ameliorate OA-induced oxidative damage and IR likely by inhibiting the expression of CYP2E1 and CYP4A, promote lipid accumulation by inducing the expression of SREBP-1c and PNPLA3 in the tested cells.
    13  Comparative Brain Pharmacokinetic Study of Jiaotai Pills in Normal and Insomnic Rats using Brain Microdialysis Combinated with LC-MS/MS
    Xiao-jun Zhang Jia-hui Chen Liang Luo Wei He Guang-hui Liu Jing Gong Yi-feng Zeng Zhi-yong Xie Qiong-feng Liao
    2018, 10(2):205-213. DOI: 10.1016/j.chmed.2018.03.006
    [Abstract](875) [HTML](0) [PDF 0.00 Byte](8)
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    Objective To compare the brain pharmacokinetics of five protoberberine-type alkaloids (i.e. berberine, palmatine, coptisine, epiberberine, and jatrorrhizine), which were the main bioactive constituents of Jiaotai Pills (JTP), in normal and insomnic rats orally administrated with the JTP. Methods The detection was conducted by a fully validated liquid chromatography-tandem mass spectrometry combinated with brain microdialysis method. Brain microdialysis probes were inserted into the hippocampus of rats. JTP extracts were administrated intragastrically and then brain microdialysates were collected at 30 min time intervals for 10 h. The separation of the five protoberberine-type alkaloids was carried out on a BDS Hypersil C18 column using a mobile phase consisting of acetonitrile and water (containing 5 mmol ammonium acetate adjusted to pH 5.0) within 4 min. The quanti?cation was performed by multiple reaction monitoring with the transitions of m/z 336.0-320.1 for berberine, m/z 352.0-336.1 for palmatine, m/z 338.0-322.1 for jatrorrhizine, m/z 336.0-320.1 for epiberberine, m/z 320.0-292.1 for coptisine and m/z 356.4-192.1 for IS. Results The lower limit of quanti?cation for five protoberberine-type alkaloids was 0.05 ng/mL. Linearity, accuracy, precision, stability and matrix effect of five analytes were all satisfactory. Five protoberberine-type alkaloids were quickly distributed in the brain. Moreover, signi?cant differences in the principal pharmacokinetic parameters such as AUC and T1/2 of the analytes were observed between two groups. Conclusion The LC-MS/MS method combinated with microdialysis is useful in the brain pharmacokinetic study of five protoberberine-type alkaloids. The results indicated that the rates of analytes absorption in insomnic rats were signi?cantly higher than those in normal rats. Besides, the protoberberine-type alkaloids could bring a direct effect on the neuron in the hippocampus.
    14  Hederacolchiside A1 suppresses proliferation of tumor cells by inducing apoptosis through modulating PI3K/Akt/mTOR signaling pathway
    Yan-Er Wang Kun Xu Wen-Hua Yue Qiong-Ming Xu Ben-Gang You Mi-Ya Zhang Zhan-Cheng Zhu Shi-Lin Yang Yan-Li Liu Kun-Ping Li
    2018, 10(2):214-221. DOI: 10.1016/j.chmed.2018.03.007
    [Abstract](528) [HTML](0) [PDF 0.00 Byte](10)
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    Hederacolchiside A1, exhibits cytostatic and cytotoxic activity against various cancer cells in vitro, however, the mechanism is not well understood. In this study, Hederacolchiside A1 from Pulsatilla chinensis was isolated and tested its anticancer activity and mechanism. Hederacolchiside A1 could inhibit proliferation of A549, SMMC-7721, BEL-7402, and MCF-7 cells by MTT assay. Investigations of apoptosis of treated cancer cells were identified in hederacolchiside A1 by flow cytometric analysis of annexin V expression. Based on the results of western blotting and JC-1 staining, hederacolchiside A1 reduced the mitochondrial membrane potential and Bcl-2 protein levels, whereas cleaved caspase-3 was higher. Furthermore, hederacolchiside A1 effectively inhibited the phosphorylations of phosphatidylinositol 3 kinase (PI3K), protein kinase B (Akt), and mammalian target of rapamycin (mTOR). In vivo study showed that hederacolchiside A1 (3.0, 4.5, and 6.0 mg/kg, ip) could significantly inhibit the weight of tumor in an H22 xenograft model. Similar inhibitory activities were observed when the compound (3.25, 7.5, and 15.0 mg/kg, ig) was tested in nude mice xenograft tumor models using human breast carcinoma MCF-7 cells. These data indicated that hederacolchiside A1 suppressed the proliferation of human tumor cells by inducing apoptosis through modulating the PI3K/Akt/mTOR signaling pathway.
    15  Protective effects of combination of Xuesaitong and aspirin on cerebral ischemia and reperfusion injury in rats
    Na-na Zhao Shi-min Tang Xin-yue Li Yong-mei Wu Yu-ting Zhou Jin-feng Shen Song Fang Xiang-bao Meng Gui-bo Sun Xiao-bo Sun
    2018, 10(2):222-229.
    [Abstract](888) [HTML](0) [PDF 0.00 Byte](7)
    Abstract:
    Objective To investigate the protective effects of the combination of Xuesaitong (XST) and aspirin on cerebral ischemia and reperfusion injury (CIRI) in rats, and further explore the underlying mechanisms. Methods A total of 150 male Sprague-Dawley (SD) rats were randomly divided into five groups with 30 rats in each group: sham group, middle cerebral artery occlusion/reperfusion (MCAO/R) model group, XST group, aspirin group, and XST + aspirin group. Rats were pretreated with XST, aspirin, or XST + aspirin for 7 d. One hour after the last administration, a model of CIRI was induced by MCAO/R. Neurological deficits were assessed using Longa’s five-point scale. Cerebral edema was detected by the measurement of brain water content. The volume of cerebral infarction was determined by 2,3,5-triphenyltetrazolium chloride (TTC) staining. The activities of superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GSH-Px), as well as levels of malonaldehyde (MDA) were detected by commercial kits. Enzyme-linked immunosorbent assay (ELISA) was used to determine the levels of interleukin-1 (IL-1β), interleukin-4 (IL-4), interleukin-6 (IL-6), interleukin-10 (IL-10), tumor necrosis factor-alpha (TNF-α), monocyte chemotactic protein 1 (MCP-1), and kynurenine in serum, cerebral cortex, and hippocampus of MCAO/R rats. The protein expression of nuclear factor erythroid 2-related factor (Nrf2), heme oxygenase-1 (HO-1), I-kappa B alpha (IκBα), and nuclear factor kappa B (NF-κB)/p65 in the cortex were analyzed by western blotting. Results Treatment of XST, aspirin, and XST + aspirin significantly alleviated the neurological deficits, cerebral edema, and cerebral infarct volume induced by MCAO/R. Treatment of XST, aspirin, and XST + aspirin also reduced MDA, IL-1β, IL-6,TNF-α, MCP-1, and kynurenine levels, and increased SOD, CAT, GSH-Px, IL-4, and IL-10 levels in serum, cerebral cortex, and hippocampus of MCAO/R rats. Furthermore, treatment of XST, aspirin, and XST + aspirin decreased the expression of nuclear NF-κB/p65 and increased the expression of IκBα, nuclear Nrf2, and HO-1. Importantly, the combination of XST and aspirin enhanced the protective effects of XST or aspirin treatment alone on CIRI in rats. Conclusion The combination of XST and aspirin significantly inhibited oxidative stress and inflammation in serum, cerebral cortex, and hippocampus of MCAO/R rats. The combination of XST and aspirin exerted more protective effects than XST or aspirin treatment alone. The combination of XST and aspirin might provide the synergistic therapeutic effects on CIRI, and deserve further clinical investigation.
    16  Chemical Constituents from Kalanchoe hybrida and their Cytotoxicity
    Ping-Chung Kuo Hsin-Yi Hung Yu-Ren Liao Yu-Yi Chan Tian-Shung Wu
    2018, 10(2):230-234. DOI: 10.1016/j.chmed.2018.03.003
    [Abstract](517) [HTML](0) [PDF 0.00 Byte](7)
    Abstract:
    Objective Kalanchoe hybrida (Crassulaceae) is naturalized throughout all the island of Taiwan, China. The preliminary bioassay-guided fractionation of the crude extract of K. hybrida exhibited that the chloroform and n-butanol fractions possessed potent cytotoxicity against MCF-7, NCI-H460, and SF-268 tumor cell lines at 50 μg/mL concentration. Therefore, K. hybrida was selected as a target and the chemical constituents from the chloroform and n-butanol fractions of the crude extracts of K. hybrida were identified. The potential constituents were examined for their cytotoxicity against the tumor cell lines. Methods A combination of conventional chromatographic techniques was performed on the crude extract of K. hybrida. The chemical structures of the purified constituents were identified on the basis of spectroscopic and spectrometric analysis. Results The purification results had led to the characterization of totally 37 compounds. The isolated compounds 1, 2, and 4-12 were examined for their cytotoxicity in vitro, and bufadienolides 4-8 and flavonol glycoside 11 displayed significant cytotoxicity towards all the tested tumor cell lines among these tested compounds. Conclusion The results indicated that these principles should be responsible for the bioactivity of corresponding partial fractions. The potential constituents could be further investigated to explore the new natural lead drugs.

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