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.

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.

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.

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 Vollera, Hui Xua, Ying Yaoa, Rudolf Bauerb, Ulrich Forstermanna, 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.

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