Danshen Chinese Medicine                                                                                                                                              

ShenYi Center of Chinese Medicine
神医草药中心
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Natural Patent Chinese Formulas

Clinical study of diabetic retinopathy treated by compound danshen dripping pills
Qi CX, Tan XH, Li QG, Wang XL.
Department of Ophthalmology, Affiliated Third Hospital, Sun Yat-sen University, Guangzhou 510630, China.
OBJECTIVE: To observe the therapeutic effect of compound danshen dripping pills in visual function recovery of diabetic retinopathy. METHODS: 42 patients(78 eyes) with diabetic retinopathy of I-III phase were divided into two groups in random. 43 eyes were treated with compound danshen dripping pills and the other 35 eyes with vitamin BI and Luding C for three months. The visual acuity, fundus, visual field, fundus fluorescein angiography and visual electro-physiology were observed. RESULTS: The visual acuity in treated group were significantly improved after treatment and had significant difference with those in control group (P < 0.05). The number of micro-hemorrage, microaneurysm and the mean defect (MD) of visual field in treated group decreased obviously after treatment and had significant difference with that in control group (P < 0.05). The latent period of P100 wave and a, b wave in treated group were shorter than control group, and the wave amplitude were higher than that in control group. CONCLUSION: The compound danshen pills can improve the visual acuity, control the micro-hemorrage and microaneurysm of fundus and have the effect in visual function recovery of diabetic retinopathy.

Clinical investigation of Compound Danshen Dripping Pills for the treatment of angina pectoris.
Guo ZhiXin, Jia Wei, Gao WenYuan, Xu ZhaoHui, Zhao LiBin, Xiao PeiGen
Research Institute of Tianjin Tasly Group Co. Ltd., Tianjin 300402, China.
Chinese Journal of Natural Medicines, 2003 (Vol. 1) (No. 2) 124-128
Compound Danshen Pill (CDP) is a herbal drug for the treatment of angina pectoris, which consists of herbal ingredients extracted from Salvia miltiorrhiza, Panax notoginseng [P. pseudoginseng var. notoginseng] and Cinnamomum camphora. The product is prepared by blending herbal extracts and excipients under thermal conditions, and dripping the mixture into an insoluble cooling liquid in which the droplets are rapidly solidified. Clinical studies revealed that CDP is similar to nitroglycerin in its ability to relieve pain rapidly. Clinical results also indicated that the long-term administration of CDP can increase the pain-relief rate and electrocardiogram improvement rate, and that its overall curative effect is superior to isosorbide dinitrate. A multi-centre clinical investigation showed that this new dosage form of composite Danshen is more effective in treating coronary heart disease than the tablet form due to the improved bioavailability.

Separation of water-soluble active components in Salvia miltiorrhiza bge. f. alba using capillary zone electrophoresis
Cui Y, Li Y, Shi W, Yang M, Zhao X, Xia Z. 2007 Sep;25(5):686-9.
Taishan Medical University
To establish a high performance capillary zone electrophoretic method for the separation of water-soluble active components, protocatechuic aldehyde (PAH), danshensu (DSS), and protocatechuic acid (PA), in Salvia miltiorrhiza bge. f. alba, many factors that can affect the resolution and the migration time, including the concentration of phosphate-borax buffer, pH value, separation voltage and detection wavelength were investigated. The optimized conditions were as follows: uncoated capillary column, 75 microm x 60.2 cm (effective length of 50 cm); column temperature, 25 degrees C; detection wavelength of the photo-diode-array detector, 210 nm; injection, 3.45 kPa x 4 s; separation voltage, 27.5 kV; running buffer, 5 mmol/L borax (pH 7.5). Under the optimized conditions, good separation of PAH, DSS and PA can be achieved in less than 8 min.

Analysis on genetic diversity of different Salvia miltiorrhiza geographical populations in China
Zhongguo Zhong Yao Za Zhi. 2007 Oct;32(19):1988-91.
Wang B, Zhang Y, Chen CB, Li XL, Chen RY, Chen L.
College of Life Sciences, Nankai University, Tianjin 300071, China.
OBJECTIVE: To research on genetic diversity of different Salvia miltiorrhiza geographical populations in China. METHOD: The genetic diversity of 27 S. miltiorrhiza geographical populations from ten provinces in China was estimated using amplified fragment length polymorphism (AFLP) markers. The data of amplified bands were analyzed by the software POPGENE and SPSS. RESULT: The ten primers employed produced a total of 528 discernable and reproduceable amplified fragments. There were 476 polymorphic brands. The percentage of polymorphic bands with in different populations was 90.15%. Genetic diversity analysis showed that Neis gene diversity (He) was 0.261 2 and Shannon's genetic diversity index (1) was 0.403 3. The coefficient of gene similarity was 0.504 0-0.789 0 between populations. The cluster map including all samples were obtained by UPGMA. In the map, there were seven cluster groups and one individual outside the groups. CONCLUSION: The genetic diversity with in different geographical population of S. miltiorrhiza in China is plentiful.

Role and mechanism of Salvia miltiorrhiza Bunge in alleviating injury induced by hypoxia/reoxygenation in HK-2 cells.
Zhongguo Dang Dai Er Ke Za Zhi. 2007 Dec;9(6):559-62.
Li MN, Dong WB, Cao M, Deng CL, Wang MY.
Department of Neonatology, Affiliated Hospital of Luzhou Medical College, Luzhou, Sichuan 646000, China.
OBJECTIVE: Salvia miltorrhiza Bunge (SMB) is a traditional Chinese herb, which is considered to promote blood flow and remove blood stasis. This study examined whether SMB can alleviate injury induced by hypoxia/reoxygenation (H/R) in human kidney proxiamal tubular cells-2 (HK-2 cells). METHODS: There were 3 experimental groups: control, H/R injury and SMB-treated H/R injury. H/R injury of HK-2 cells was induced by first covering the cells with and then removing liquid paraffin wax. Different concentrations of compound SMB solution (0.05%, 0.10%, 0.15% or 0.20%) were administered to the SMB-treated H/R injury group before the hypoxic injury. After 4, 12 and 24 hrs of hypoxia and 4, 12, 24 and 48 hrs of reoxygenation, morphologic changes of HK-2 cells were observed under an inverted microscope. Cell viability was measured by the MTT method. Lactate dehydrogenase (LDH) activity in the culture supernatants was assayed using biochemical methods; TNF-alpha levels were determined by radioimmunoassay (RIA). RESULTS: The number of HK-2 cells was significantly reduced in the H/R injury group after hypoxia, and reached a nadir 24 hrs after hypoxia treatment. Various concentrations of SMB-treated groups showed significantly greater number of HK-2 cells than the H/R injury group. SMB solution (0.10%) produced the best effect. The levels of LDH and TNF-alpha in the H/R injury group were significantly increased, and reached a peak between 24 hrs of hypoxia and 4 hrs of reoxygenation when compared to the control group. Pre-treating with 0.10% SMB resulted in significantly lower levels of LDH and TNF-alpha than in the untreated H/R injury group at various time points of H/R. CONCLUSIONS: SMB has protective effects against H/R injury of HK-2 cells, possibly through inhibition of inflammatory cytokines.

 Enhanced antioxidant bioactivity of Salvia miltiorrhiza (Chinese Medicine Danshen) products prepared using nanotechnology.
Liu JR, Chen GF, Shih HN, Kuo PC.
Phytomedicine. 2008 Jan;15(1-2):23-30.
Department of Animal Science and Technology and Institute of Biotechnology, National Taiwan University, Taipei 102, Taiwan, ROC.
The traditional Chinese medicine, Salvia miltiorrhiza (Danshen), promotes blood circulation and relieves blood stasis, also demonstrating good antioxidant activity. In the present study, therefore, the antioxidant activities of medicinal plant materials prepared using nanotechnology or traditional grinding methods were compared using three biological assays. It was found that the nanotechnology preparation had stronger antioxidant bioactivities. Complementary quantitative analysis of four active constituents, salvianolic acid B, cryptotanshinone, tanshinone I and tanshinone IIA, by HPLC revealed only marked differences for salvianolic acid B. The results indicate that the polar active constituent in the nanotechnology samples was released faster compared to the traditionally powdered samples.

Salvia miltiorrhiza treatment during early reperfusion reduced postischemic myocardial injury in the rat.
Nie R, Xia R, Zhong X, Xia Z.
Can J Physiol Pharmacol. 2007 Oct;85(10):1012-9.
Department of Cardiology, Second Affiliated Hospital, Sun Yat-sen University, Guangzhou, Guangdong, China.
Oxidative stress may play a causative role in myocardial ischemia-reperfusion injury. However, it is a relatively understudied aspect regarding an optimal timing of antioxidant intervention during ischemia-reperfusion. The present study investigates the effect of different treatment regimens of Salvia miltiorrhiza (SM) herb extracts containing phenolic compounds that possess potent antioxidant properties on postischemic myocardial functional recovery in the setting of global myocardial ischemia and reperfusion. Langendorff-perfused rat hearts were subjected to 40 min of global ischemia at 37 degrees C followed by 60 min of reperfusion, and were randomly assigned into the untreated control and 2 SM-treated groups (n = 7 per group). In treatment 1 (SM1), 3 mg/mL of water soluble extract of SM was given for 10 min before ischemia and continued during ischemia through the aorta at a reduced flow rate of 60 muL/min, but not during reperfusion. In treatment 2 (SM2), SM (3 mg/mL) was given during the first 15 min of reperfusion. During ischemia, hearts in the control and SM2 groups were given physiological saline at 60 muL/min. The SM1 treatment reduced the production of 15-F2t-isoprostane, a specific index of oxidative stress-induced lipid peroxidation, during ischemia (94 +/- 20, 43 +/- 6, and 95 +/- 15 pg/mL in the coronary effluent in control, SM1, and SM2 groups, respectively; p < 0.05, SM1 vs. control or SM2) and postponed the onset of ischemic contracture. However, SM2, but not the SM1 regimen, significantly reduced 15-F2t-isoprostane production during early reperfusion and led to optimal postischemic myocardial functional recovery (left ventricular developed pressure 51 +/- 4, 46 +/- 4, and 60 +/- 6 mmHg in the control, SM1, and SM2 groups, respectively, at 60 min of reperfusion; p < 0.05, SM2 vs. control or SM1) and reduced myocardial infarct size as measured by triphenyltetrazolium chloride staining (26% +/- 2%, 22% +/- 2%, and 20% +/- 2% of the total area in the control, SM1, and SM2 groups, respectively, p < 0.05, SM2 vs. control). It is concluded that S. miltiorrhiza could be beneficial in the treatment of myocardial ischemic injury and the timing of administration seems important.

 Ameliorating effects of compounds derived from Salvia miltiorrhiza root extract on microcirculatory disturbance and target organ injury by ischemia and reperfusion.
Han JY, Fan JY, Horie Y, Miura S, Cui DH, Ishii H, Hibi T, Tsuneki H, Kimura I.
Pharmacol Ther. 2007 Oct 16
Department of Integration of Chinese and Western Medicine, School of Basic Medical Sciences, Peking University, Beijing, China; Tasly Microcirculation Research Center, Peking University Health Science Center, Beijing, China; Department of Internal Medicine, School of Medicine, Keio University, Tokyo, Japan.
Ischemia and reperfusion (I/R) exerts multiple insults in microcirculation, frequently accompanied by endothelial cell injury, enhanced adhesion of leukocytes, macromolecular efflux, production of oxygen free radicals, and mast cell degranulation. Since the microcirculatory disturbance results in injury of organ involved, protection of organ after I/R is of great importance in clinic. Salvia miltiorrhiza root has long been used in Asian countries for clinical treatment of various microcirculatory disturbance-related diseases. This herbal drug contains many active water-soluble compounds, including protocatechuic aldehyde (PAl), 3,4-dihydroxyphenyl lactic acid (DLA) and salvianolic acid B (SalB). These compounds, as well as water-soluble fraction of S. miltiorrhiza root extract (SMRE), have an ability to scavenge peroxides and are able to inhibit the expression of adhesion molecules in vascular endothelium and leukocytes. Moreover, lipophilic compounds of SMRE also prevent the development of vascular damage; NADPH oxidase and platelet aggregation are inhibited by tanshinone IIA and tanshinone IIB, respectively, and the mast cell degranulation is blunted by cryptotanshinone and 15,16-dihydrotanshinone I. Thus, the water-soluble and lipophilic compounds of SMRE appear to improve the I/R-induced vascular damage multifactorially and synergically. This review will summarize the ameliorating effect of compounds derived from SMRE on microcirculatory disturbance and target organ injury after I/R and will provide a new perspective on remedy with multiple drugs.

Physicochemical properties and drug release rate of calcium phosphate cement containing danshen composite injection
Jiang H, Huang X, Tan Y.
Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi. 2007 Oct;21(10):1113-7.
Institute of Composite Material, Tianjin University, Tianjin 300072, PR China.
OBJECTIVE: To investigate the physicochemical properties of the calcium phosphate cement (CPC) containing Danshen composite injection and its drug release rate. METHODS: This experiment included 4 groups and each group contained 6 specimens. CPC (2 g) was mixed with the setting solution that served as the control group; 0.1, 0.5 and 1.0 ml of Danshen composites injection (concentration, 1 000 mg/ml; pH, 7.35) were respectively added to CPC (2 g), which were used as the experimental groups 1, 2 and 3. The resulting specimens were investigated by the X-ray diffraction (XRD), the fourier transformed infrared spectroscopy (FTIR), and the scanning electron microscope (SEM). RESULTS: The XRD analysis showed that the control group had a typical diffraction pattern of the hydroxypatite (HAP), which was consistent with the standard pattern of HAP. When more Danshen was added in the experimental groups, the diffraction peaks of HAP gradually decreased; when the diffraction angle 2theta was about 25.92 degrees, the HAP peaks disappeared. Based on the FTIR analysis, with an increase of the drug concentration, the absorption peak of the hydroxy groups decreased. The SEM showed that the size of the CPC particle was related to the drug concentration; with an increase of the drug concentration, the CPC particle increased in number, resulting in an increasing trend of coacervation. The elution test showed that the drug-release rate and capacity varied with the different concentrations of Danshen medicine. The initial release rate was relatively great, but after 96 hours the rate slowed down, lasting for a long time. CONCLUSION: The physicochemical properties of CPC do not change when a proper dose (0.1 ml/2 g) of Danshen is added to CPC. The Danshen composite can be effectively released from CPC, and so CPC can be used as an ideal drug-delivery carrier for Danshen composite.

Functional genomics studies of Salvia miltiorrhiza III. analyze of metallothionein (MT-2) genes
Cui GH, Mao Y, Huang LQ, Yuan Y, Wang XY.
Zhongguo Zhong Yao Za Zhi. 2007 Jul;32(14):1393-5.
Institute of Chinese Materia Medica, Academy of Chinese Medical Sciences, Beijing 100700, China.
OBJECTIVE: To study the metallothionein genes of Salvia miltiorrhiza through bioinformatics and characterization of tissue expression in regenerated shoots. METHOD: Metallothionein genes were obtained by cDNA microarray analyze. BLAST was used for align, ORF finder software was used to find open reading frame, Prosite database was used to analyze the protein. Semi-quantitative RT-PCR method was used to detect the gene expression level. RESULT: Two metallothionein genes were obtained which were contained a deduced amino acid sequence of 80 and 79 residues, named as SmMT-2a and SmMT-2b, they had a homology of 71.25%. Semiquantitative RT-PCR indicated that metallothionein genes were expressed in all tissues such as root, stem and leaf in regenerated shoots, while the expression level was higher in leaf than in root and stem. CONCLUSION: It was the first time that metallothionein genes were obtained from S. miltiorrhiza. It provides a good basis for further functional study of S. miltiorrhiza.

Cardiovascular effects of Danshen
Tsung O. Cheng,
Department of Medicine, The George Washington University Medical Center, 2150 Pennsylvania Avenue, N.W., Washington, D.C. 20037, United States
Danshen is one of the most versatile Chinese herbal drugs that have been used for hundred of years in the treatment of numerous ailments. Because of its properties of improving microcirculation, causing coronary vasodilatation, suppressing the formation of thromboxane, inhibiting platelet adhesion and aggregation, and protecting against myocardial ischemia, it is widely used either alone or in combination with other herbal ingredients for patients with coronary artery disease and other cardiovascular diseases, in both China and other countries including the United States. This article provides an overview of its history, pharmacology, pharmacokinetics, clinical applications, side effects, interactions with Western drugs, and future prospects in the management of cardiovascular diseases.

Ursolic acid from the Chinese medicine Danshen (Salvia miltiorrhiza L.) upregulates eNOS and downregulates Nox4 expression in human endothelial cells
Katja Steinkamp-Fenskea, Larissa Bollingera, Natalie Völlera, Hui Xua, Ying Yaoa, Rudolf Bauerb, Ulrich Förstermanna, Huige Lia,
a. Department of Pharmacology, Johannes Gutenberg University, Obere Zahlbacher Strasse 67, D-55131 Mainz, Germany
b. Department of Pharmacognosy, Karl Franzens University, Graz, Austria
Danshen, the dried root of Salvia miltiorrhiza Bunge (Lamiaceae), is one of the most commonly used traditional Chinese medicines for cardiovascular indications. In EA.hy 926 cells, a cell line derived from human umbilical vein endothelial cells (HUVEC), an aqueous extract of Danshen, and also a methanol extract of the plant, increased eNOS promoter activity, eNOS mRNA and protein expression, as well as endothelial NO production. A dichloromethane extract, in contrast, did not change eNOS gene expression. Thus, the active Danshen constituent(s) responsible for eNOS upregulation is (are) hydrophilic and/or alcohol-soluble. One such compound is ursolic acid that significantly increased eNOS expression in EA.hy 926 cells and native HUVEC, and enhanced bioactive NO production measured in terms of its cGMP increasing activity. Other tested hydrophilic and alcohol-soluble compounds isolated from Danshen had no effect on eNOS expression. Interestingly, ursolic acid also reduced the expression of the NADPH oxidase subunit Nox4 and suppressed the production of reactive oxygen species in human endothelial cells. Upregulation of eNOS and a parallel downregulation of Nox4 lead to an increase in bioactive NO. This in turn could mediate some of the beneficial effects of Danshen. Ursolic acid is a prototypical compound responsible for this effect of the plant.

Relaxant effects of danshen aqueous extract and its constituent danshensu on rat coronary artery are mediated by inhibition of calcium channels
Francis Fu Yuen Lam, a, , John Hok Keung Yeunga, Kam Ming Chana and Penelope Mei Yu Ora
Department of Pharmacology, Faculty of Medicine, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong
In this study, we have investigated the actions of danshensu, an active, water-extractable component of the medicinal herb danshen (Salvia miltiorrhiza), on rat isolated coronary artery rings precontracted with 1 μM 5-hydroxytryptamine (5-HT) and its action compared to the water-extractable fraction of the herb. Extraction of the water-soluble fraction from danshen (S. miltiorrhiza) provided yield of 17.5% (35 g/200 g). The amount of danshensu determined in the crude danshen herb and in its aqueous fraction was 0.45 mg/g (0.045%) and 3.28 mg/g (0.33%). The danshen aqueous extract was 13 times less potent than danshensu in relaxing 5-HT-precontracted coronary artery rings; IC50 values were 930.3 ± 133.5 μg/ml and 71.5 ± 11.0 μg/ml. Removal of the endothelium did not significantly affect their vasodilator potencies; IC50 values were 842.1 ± 123.8 μg/ml and 84.8 ± 8.8 μg/ml. On the other hand, a potassium channel inhibitor tetraethylammonium (TEA, 10 mM) shifted their concentration–response curves by 1.7 and 2.2 folds. The possible involvement of Ca2+ channels was investigated in artery rings incubated with Ca2+-free buffer and primed with 1 μM 5-HT or 60 mM KCl for 5 min prior to addition of CaCl2 to elicit contraction. In 5-HT-primed preparations, the CaCl2-induced vasoconstriction was abolished by 2 mg/ml danshen aqueous extract and 200 μg/ml danshensu, whereas, in KCl-primed preparations, 10 mg/ml danshen aqueous extract and 600 μg/ml danshensu were required to abrogate the vasoconstriction. These findings suggest the vasorelaxant actions of danshen aqueous extract and danshensu were produced by inhibition of Ca2+ influx in the vascular smooth muscle cells. The opening of K+ channels had a minor contribution to the response, but endothelium-dependent mechanisms were not involved.

Danshen: A versatile Chinese herbal drug for the treatment of coronary heart disease
Tsung O. Chenga,
aGeorge Washington University, Washington, DC 20037

Danshen: What every cardiologist should know about this Chinese herbal drug
Tsung O. Chenga,
aGeorge Washington University, Washington, DC 20037, United States

Danshen: A Popular Chinese Cardiac Herbal Drug
Tsung O. Cheng MD, , FACC
Journal of the American College of Cardiology, Volume 47, Issue 7, 4 April 2006, Pages 1499-1500
John H.K. Vogel

Discriminating the Genuineness of Chinese Medicines Using Least Squares Support Vector Machines
Ke Yua and Yiyu Cheng, a,
a.Pharmaceutical Informatics Institute, Zhejiang University, Hangzhou 310027, China
A method for the rapid identification of the genuineness of Chinese medicines based on near infrared (NIR) spectroscopy and least squares support vector machines (LSSVM) was proposed. In this study, NIR spectra of the powdered Danshen (Radix Salviae Miltiorrhizae) were collected, and the nonlinear classifier based on LSSVM algorithm was developed to discriminate the genuineness of these herbs. The result obtained by the proposed method was compared with those from the traditional support vector machines (SVM) and BP-ANN methods. It was shown that the generalization performance of the classifier based on LSSVM was much better than that of BP-ANN, and the computation time of LSSVM was much shorter than that of the traditional SVM. The proposed method could be applied to the rapid and accurate identification of the quality of the natural products.

Chinese herbs Danshen and Gegen modulate key early atherogenic events in vitro
Daniel P. Sievekinga, , Kam-Sang Woob, Kwok P. Fungb, Pia Lundmana, Shirley Nakhlaa and David S. Celermajera, c, d, ,
a.Heart Research Institute, 145 Missenden Rd, Camperdown, Sydney, NSW, Australia, 2050
b.The Chinese University of Hong Kong, Hong Kong
c.Royal Prince Alfred Hospital Sydney, NSW, Australia
d.Department of Medicine, University of Sydney, Australia
Danshen (Salvia miltiorrhiza) and Gegen (Radix puerariae) are two herbs used in traditional Chinese medicine, most commonly for their putative cardioprotective and anti-atherosclerotic effects. In this study, we investigated the effect of a preparation of these herbs on two key processes in the early stages of atherosclerosis; macrophage lipid loading and monocyte adhesion to endothelial cells.
Human monocyte derived macrophages (HMDMs) were treated with 0.1–1.0 mg/ml of the herbal mixture in aqueous buffers and loaded with acetylated LDL (AcLDL) (50 μg/ml) for 72 h, and analyzed for cholesterol (C) and cholesteryl esters (CE), via HPLC. Human endothelial cell monolayers were also treated with 0.1–1.0 mg/ml of the herbal mixture and monocyte adhesion measured. Cell adhesion molecules E-selectin, ICAM-1 and VCAM-1 were assessed via ELISA.
Compared to control conditions, the herbal mixture induced a significant dose-related decrease in the total cholesterol (free and esterified) in the HMDMs (p<0.001 by ANOVA). By contrast, the herbs also induced an increase in ICAM-1 expression (p<0.001) and monocyte adhesion at higher concentrations (p<0.05).
In conclusion, treatment of cells with this preparation of Danshen and Gegen, a commonly used Chinese health supplement, results in a dose-related suppression of AcLDL uptake by human macrophages, and an increase in the level of ICAM-1 expression and adhesion of monocytes to endothelial cells. These herbs therefore show the ability to modulate key early events in atherosclerosis.

Magnesium as NMDA receptor blocker in the traditional Chinese medicine Danshen
X. Suna, b, c, L.N. Chana, b and N.J. Suchera, b, d, ,
a.Department of Biology, Hong Kong University of Science & Technology, Clear Water Bay, Kowloon, Hong Kong SAR, China
bMolecular Neuroscience Center, Hong Kong University of Science & Technology, Clear Water Bay, Kowloon, Hong Kong SAR, China
c.Institute of Neuroscience, Nanjing Medical University, Nanjing, Jiangsu, China
d.Biotechnology Research Institute, Hong Kong University of Science & Technology, Clear Water Bay, Kowloon, Hong Kong SAR, China
Aqueous extracts of the traditional Chinese medicine Danshen, the dried roots of Salvia miltiorrhiza Bunge (Labiatae), blocked N-methyl-d-aspartate (NMDA) evoked currents in cerebrocortical neurons in vitro. The block of the NMDA-evoked currents was voltage dependent and showed the negative slope conductance reminiscent of the effect of Mg2+ ions. Atomic absorption spectrophotometry (AAS) revealed that aqueous Danshen extracts contained 9 mM magnesium. Fractionation of the extracts by high performance liquid chromatography followed by patch clamp recording and AAS indicated that magnesium ions were present in two distinct fractions. One fraction contained 5 mM magnesium and blocked NMDA-induced currents indicating that it contained mostly free Mg2+ ions, while a second fraction did not possess NMDA antagonist activity despite the presence of 4 mM magnesium suggesting that Mg2+ in this fraction was mostly chelated. Following removal of the free Mg2+ by ion exchange chromatography, the previously observed block of the NMDA-induced currents was abolished. These data demonstrate that Danshen contains both free and chelated Mg2+. Free Mg2+ ions account for the NMDA antagonist activity of Danshen in vitro.

Protective effects of Danshensu from the aqueous extract of Salvia miltiorrhiza (Danshen) against homocysteine-induced endothelial dysfunction
K. Chana, S.H. Chuia, D.Y.L. Wonga, W.Y. Haa, C.L. Chana and R.N.S. Wonga, b, ,
a.Research and Development Division, School of Chinese Medicine, Hong Kong Baptist University, Hong Kong
b.Department of Biology, Science Faculty, Hong Kong Baptist University, Hong Kong
Homocysteine (Hcy) is a by-product of methionine metabolism. An imbalance of Hcy in the body may lead to hyperhomocysteinemia, a condition with elevated Hcy concentration in blood that may be one of the risk factors responsible for the development of several vascular diseases (thromboembolism, atherosclerosis, stroke, vascular diseases and dementia). Radix Salvia miltiorrhiza (Danshen), a well-known Chinese medicinal herb that can activate and improve blood microcirculation, is noticeable for its beneficial effect in treating cardiovascular diseases. The present study is to demonstrate the protective effect of Danshen extract against the homocysteine-induced adverse effect on human umbilical vein endothelial cell (HUVEC). Homocysteine (5 mM) not only decreased the cell viability but also caused the disruption of capillary-like structure formation in vitro. The protective effect of Danshen aqueous extract and its active compounds on endothelial cell function were demonstrated through an in vitro tube formation assay, which mimics the new blood vessel formation. To identify the active components in the aqueous extract of Danshen, the content was characterized by instrumental analysis using high performance liquid chromatography with diode array detector (DAD) and electrospray tandem mass spectrometry (ESI-MS/MS). Interestingly, Danshen extract and its pure compounds showed different effectiveness in protecting HUVEC against Hcy-induced injury according to the following descending order: Danshen aqueous extract, 3-(3,4-dihydroxy-phenyl)-2-hydroxy-propionic acid (Danshensu), protocatechuic acid, catechin and protocatechualdehyde. We believed that such findings might provide evidence in understanding the beneficial effects of Danshen on the cardiovascular system.

Study of the determination and pharmacokinetics of Compound Danshen Dripping Pills in human serum by column switching liquid chromatography electrospray ion trap mass spectrometry
Wei-Jing Peia, Xin-Feng Zhaob, Zhong-Min Zhuc, Chang-Zheng Lind, Wen-Ming Zhaoa and Xiao-Hui Zhengc, ,
a.Department of Biology Engineering, School of Life Science and Technology, Xi’an Jaotong, University, Xi’an, ShaanXi, China
b.Science School of Xi’an Jiaotong University, Xi’an, ShaanXi, China
c.Engineering and Research Center of Chinese Herb Modernization, Xi’an Jiaotong University, Xi’an, ShaanXi, China
d.Pharmacy Department of Second Hospital, Xi’an Jiaotong University, Xi’an, ShaanXi, China
Compound Danshen Dripping Pill (CDDP) is an important drug widely used for the treatment of cardiovascular diseases. An active component Danshensu (DS) of CDDP was separated by reversed-phase high performance liquid chromatography using column-switching system and analyzed by electrospray mass spectrometry. With this validated assay the pharmacokinetics of CDDP was studied in 10 healthy volunteers after a single oral administration of 250 mg. After trichloroacetic acid precipitation of serum proteins, the analytes were preconcentrated and black-flushed on a reversed-phase column for separation using a switching valve. The analytes were ionized using negative electrospray ionization (ESI) mode. The precursor ion of m/z 196.6 was used to quantify DS in serum. The linear calibration curve ranged from 1.25 to 175 μg/mL. The limit of quantification (LOQ) for DS was 0.15 μg/mL. The intra-day and inter-day precision (R.S.D.) was less than 7.4 and 7.9%, respectively.

Cardiovascular pharmacotherapy and herbal medicines: the risk of drug interaction
Angelo A. Izzo, a, , Giulia Di Carloa, Francesca Borrellia and Edzard Ernstb
a.Department of Experimental Pharmacology, University of Naples “Federico II”, via D. Montesano 49, 80131 Naples, Italy
b.Complementary Medicine, Penninsula Medical School, Universities of Exeter and Plymouth, 25 Victoria Park Road, EX2 4NT, UK
Use of herbal medicines among patients under cardiovascular pharmacotherapy is widespread. In this paper, we have reviewed the literature to determine the possible interactions between herbal medicines and cardiovascular drugs. The Medline database was searched for clinical articles published between January 1996 and February 2003. Forty-three case reports and eight clinical trials were identified. Warfarin was the most common cardiovascular drug involved. It was found to interact with boldo, curbicin, fenugreek, garlic, danshen, devil's claw, don quai, ginkgo, papaya, lycium, mango, PC-SPES (resulting in over-anticoagulation) and with ginseng, green tea, soy and St. John's wort (causing decreased anticoagulant effect). Gum guar, St. John's wort, Siberian ginseng and wheat bran were found to decrease plasma digoxin concentration; aspirin interactions include spontaneous hyphema when associated with ginkgo and increased bioavailability if combined with tamarind. Decreased plasma concentration of simvastatin or lovastatin was observed after co-administration with St. John's wort and wheat bran, respectively. Other adverse events include hypertension after co-administration of ginkgo and a diuretic thiazide, hypokalemia after liquorice and antihypertensives and anticoagulation after phenprocoumon and St. John's wort. Interaction between herbal medicine and cardiovascular drugs is a potentially important safety issue. Patients taking anticoagulants are at the highest risk.

A taste of Chinese medicine!
Mohammad Bashar Izzat, FRCS(CTh), , a, Anthony P. C. Yim MDa and M. Hazem El-Zufari MDa
a.Division of Cardiothoracic Surgery, Department of Surgery, Prince of Wales Hospital, The Chinese University of Hong Kong, Hong Kong
We report a case of profound anticoagulation caused by interaction between warfarin and danshen, a widely used Chinese herbal medicine, in a patient who had undergone mitral valve replacement. Patients taking warfarin should be warned not to take this herb. In addition, physicians should be alert to the possibility of an interaction with herbal medicine when anticoagulation control becomes difficult and no other causes are apparent.

Herb-drug interactions
Adriane Fugh-Berman MDa,
aGeorge Washington University School of Medicine and Health Sciences, Department of Health Care Sciences, 2150 Pennsylvania Avenue, NW 2B-417, Washington, DC 20037, USA
The Lancet, Volume 355, Issue 9208, 18 March 2000, Page 1020
Concurrent use of herbs may mimic, magnify, or oppose the effect of drugs. Plausible cases of herb-drug interactions include: bleeding when warfarin is combined with ginkgo (Ginkgo biloba), garlic (Allium sativum), dong qual (Angelica sinensis), or danshen (Salvia miltiorrhiza); mild serotonin syndrome in patients who mix St John's wort (Hypericum perforatum) with serotonin-reuptake inhibitors; decreased bioavailability of digoxin, theophylline, cyclosporin, and phenprocoumon when these drugs are combined with St John's wort; induction of mania in depressed patients who mix antidepressants and Panax ginseng; exacerbation of extrapyramidal effects with neuroleptic drugs and betel nut (Areca catechu); increased risk of hypertension when tricyclic antidepressants are combined with yohimbine (Pausinystalla yohimbe); potentiation of oral and topical corticosteroids by liquorice (Glycyrrhiza glabra); decreased blood concentrations of prednisolone when taken with the Chinese herbal product xaio chai hu tang (sho-saiko-to); and decreased concentrations of phenytoin when combined with the Ayurvedic syrup shankhapushpi. Anthranoid-containing plants (including senna [Cassia senna] and cascara [Rhamnus purshiana]) and soluble fibres (including guar gum and psyllium) can decrease the absorption of drugs. Many reports of herb-drug interactions are sketchy and lack laboratory analysis of suspect preparations. Health-care practitioners should caution patients against mixing herbs and pharmaceutical drugs.

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