Traditional Chinese medicine of Salvia miltiorrhiza Bunge: a review of phytochemistry, pharmacology and pharmacokinetics

Salvia miltiorrhiza Bunge,known as red sage or Danshen,is commonly applied in Chinese herbal therapeutics for various biological effects.In 25-220 C.E.,the earliest Chinese Materia Medica referred to as Shengong’s Classic of Materia Medica(Eastern Han Dynasty of China,200 C.E.)initially documented Salvia miltiorrhiza Bunge.Currently,Salvia miltiorrhiza Bunge is one of the most frequently prescribed traditional Chinese medicines in clinics for antidiabetic,antianxiety,antihypertensive,antidepressant,antilipidemic,antiarrhythmic,and anticancer effects.Undoubtedly,it is best known for improving and promoting blood circulation;thus,its recommendation for use in cardiovascular disorders.Regarding this article,important information was sourced both manually and electronically using patent compendium and databases,including Google Scholar,herbal plant monographs,Hindawi,PubMed,Embase,CNKI,pharmacopoeias,Springer,etc.Well-concise details and discussions on the botany,traditional medicinal uses,chemical constituents-phytochemistry,pharmacokinetics,pharmacology,and toxicology information are presented.In perspective,understanding the comprehensive information of the above-stated fields with regards to Salvia miltiorrhiza Bunge would yield an additional positive knowledge on areas that need urgent attention and identify gaps in research and safety for future consideration.

PRIMARY STUDY ON MECHANISM OF COLLAGEN SYNTHESIS INHIBITION IN CULTURED HUMAN FETAL HEPATOCYTES PRETREATED BY SALVIA MILTIORRHIZA BUNGE

Objective The effects of salvia miltiorrhiza bunge (SMB) on collagen synthesis in the human fetal hepatocytes culture were studied. Methods The collagen synthesis of hepatocytes were stimulated by the addition of carbon tetrachloride (CCl 4 ) to the culture medium, the concentration of type procollagen (PC) in the culture medium and the hydroxyproline (Hyp) in hepatocytes were determined, as well as the activity of se dependent glutathione peroxidase (Se GSH Px) and the concentration of malondiadehyde (MDA) in the culture medium. Results A significant decrease in PC, Hyp and MDA production, and the significant increase in Se GSH Px activity were observed in the cultures pretreated with 1 g L -1 SMB for 4 hours compared with the untreated cultures. Analysis of the Se GSH Px/MDA ratio in SMB pretreated group showed more marked increase compared to that of the untreated group ( P <0.01). There was a significant negative correlation between the ratio of Se GSH Px/MDA and the concentration of PC in SMB pretreated group ( r=-0.9017, P <0.01). Conclusion Our results indicate that SMB may suppress the collagen synthesis of cultured human fetal hepatocytes stimulated by CCl 4 , and its mechanism may be related to the increase in Se GSH Px/MDA ratio and the enhancement of hepatocytes antioxidation capability.

Published papers about Salvia miltiorrhiza Bunge as a Chinese medicinal material

Xu Cuihong, Shu Zhiming, Wang Yan, Miao Fang, Zhou ke. The accumulation rule of the main medicinal components in different organs of Salvia miltiorrhiza Bunge. and Salvia miltiorrhiza Bunge. f. alba. Lishizhen Medicine and Materia Medica Research, 2010, 21（09）： 2129-2132. Email： miaofangmf@ 163,com

EFFECTS OF SALVIA MILTIORRHIZA BUNGE （SMB） ON LIPID PEROXIDATION OF CULTURED HUMAN FATAL HEPATOCYTES

The role of salvia miltiorrhiza bunge (SMB) In protection of lipid peroxidation induced by CCl4 was studied in primary cultured human fetal hepatocytes. The results showed that SMB had significant effect inhibiting lipid peroxidation of the hepatocytes when exposed to CCl4 as compared with the non-protective co4trol. SMB could decrease the production of malondialdehyde (MDA) and the release of ALT, and increase the synthesis of albumin (ALB) and the activity of Se-dependent glutothione peroxidase (Se-GSH-Px). These results suggest that the suppression of lipid peroxidation may be largely due to the increased activity of Se-GSH-Px.

Published papers about Salvia miltiorrhiza Bunge used in traditional Chinese medicine

Chu Kedan, Ji Guolu, Shi Hong, Xie Weirong, Zhang Yuqin, Zhang Xiaoqin. Study of the alcohol extraction process of Shenhu granules. Strait Pharmaceutical Journal, 2011, 23（03）： 21-24. Email： chukd5917@ 163.com

EFFECTS OF SALVIA MILTIORRHIZA BUNGE（SMB）ON LIPID PEROXIDATION AND PLASMA MEMBRANE FLUIDITY OF CULTU

In order to investigate the effect of salvia miltiorrhiza hunge(SMB)on the plasma membrane fluidity and the relationship between the lipid peroxidation and the Plasma membrane fluidityin cultured human fetdal hepatocytes,the plasma membrane fluidity,using 1,6-dipheny-1,3,5-hexatriene(DPH)as a fluorescence probe, malondialdehyde(MDA)production as well as alanine aminotransferase(ALT)release of human fetal hepatocytes cultured in Presence of carbon tetrachloride(CCl4)or SMB puls CCl4 were estimated. In the cultured hepatocytes injured by CCl4,significant increments of the MDA production and the ALT release,and significant decrease in the plasma membrane fluidity were observed.when the culture medium was supplied with SMB prior to the additionof CCl4,the CCl4 induced increments in MDA production and ALT release was suppressed signifi cantly and a concomitant raise of plasma membrane fluidity towards normal occurred.The resultssuggested that SMB could suppress the lipid peroxidation in bepatocytes,thereby normal membranefluidity might be retained.

Molecular mechanism of Salvia miltiorrhiza Bunge and Carthamus tinctorius L. in the treatment of myocardial ischemia based on network pharmacology: a comparative study

Based on network pharmacology and molecular docking technology,the similarities and differences of anti-myocardial ischemia mechanism between Salvia miltiorrhiza Bunge(SM)and Carthamus tinctorius L.(CT)were studied.Firstly,based on the traditional Chinese medicine systems pharmacology(TCMSP),the related compounds of SM and CT were obtained,and the potential targets of these compounds were collected by the target fishing method.Genecards database was used to obtain targets related to myocardial ischemia.The cross targets of CT,SM,and myocardial ischemia were then selected,and the protein-protein interaction(PPI)network was constructed based on the STRING database.The cross targets were imported into the Metascape database for Gene Oncology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)analysis.Cytoscape software was used to build the topological network diagram of the drug-compound-target path.Finally,the binding ability of the active ingredient and the key target was verified by molecular docking.65 active ingredients and 38 potential targets were screened from SM,and 22 active ingredients and 58 potential targets were excavated from CT.Important targets common to SM and CT were TNF,IL6,VEGFA,AKT1,etc.The common enrichment pathways involved are fluid shear stress and atherosclerosis,IL-17 signaling pathway,pathways in cancer,and toxoplasmosis.The findings suggested that the two traditional Chinese medicines exerted the effect of myocardial ischemia through the characteristics of multiple targets,multiple pathways,and multiple compounds.

The anti-pulmonary fibrosis of Salvia miltiorrhiza Bunge: A systematic review

The characteristics of PF are diffuse alveolitis and disruption of alveolar structure leading to pulmonary interstitial fibrosis.At the same time,pulmonary fibrosis reduces lung volume and restricts ventilation,ultimately leading to hypoxemia and respiratory failure.In clinical guidelines,Nintedanib or Pirfenidone is often used for treatment.However,the two drugs,although they may slow the progression of the disease,cannot stop,or reverse fibrosis and generally result in a variety of toxic side effects.In 184-220 A.D.,it was already recorded in Supplementary Records of Famous Physicians that the function of SMB was to invigorate blood circulation and disperse blood stasis.In modern medical research,the active ingredients of SMB have likewise been found to be used in treatments such as anti-fibrosis,anti-inflammatory,antithrombotic,antioxidant,microcirculatory improvement,and antineoplastic.In this review,a comprehensive search of the former literature on SMB and pulmonary fibers was conducted using databases including PubMed,CNKI,the National Science and Technology Library,Hindawi,Chinese Science and Technology Journal Database,and the Scientific Network Database.Meanwhile,this review presents the mechanisms of the active ingredients in SMB which have anti-pulmonary fibrosis effects through the signal pathways,cytokines,inflammation response,oxidative stress,apoptosis,and matrix metalloproteinases.SMB could offer a direction for therapy for pulmonary fibrosis by identifying the possible active components and exploring the potential mechanisms.Those could provide a reference for further research and application of SMB in the treatment of pulmonary fibrosis.

Studies on the comparative pharmacokinetic of three phenolic acids of Salvia miltiorrhiza Bunge in normal and hyperlipidemic rats by liquid chromatography-tandem mass spectrometry

The hydrophilic phenolic acid is the main component of Salvia miltiorrhiza Bunge against cardiovascular diseases.A validated and sensitive liquid chromatography-tandem mass spectrometry(LC-MS/MS)method was established to simultaneously determinate three phenolic acids(danshensu,protocatechuic acid and rosmarinic acid)in rats to investigate their pharmacokinetic profiles firstly.The values for limit of quantification were in range of 0.25-1.25μg/L.Meanwhile,the suitable intra-day and inter-day precision as well as extraction recoveries were also obtained.And such an established method was finally successfully applied to compare the pharmacokinetic behaviors of three phenolic acids in normal and hyperlipidemic rats after oral administration of Salvia miltiorrhiza Bunge.The results showed there were significant differences in pharmacokinetic parameters between normal and hyperlipidemic groups.In especial,the plasma level of three phenolic acids in hyperlipidemic groups was significantly higher than normal ones.This comparative pharmacokinetic study provided direct evidence for improving clinical rational use of Salvia miltiorrhiza Bunge.

Chemical constituents and pharmacological activities of Salvia miltiorrhiza

Salvia miltiorrhiza Bunge(S.miltiorrhiza),a perennial plant of the genus Salvia,is widely used in traditional folklore medicine.Previous chemical research on this plant contains diterpenoid quinones,phenolic acids,polysaccharides and other compounds.The pharmacological investigation of S.miltiorrhiza has shown that it has various pharmacological activities,such as cardiovascular system protection,anti-inflammatory,anti-oxidant,anti-tumor,liver protection,and neuroprotection activities.This research tends to give an overview of the main chemical constituents and pharmacological effects of S.miltiorrhiza,aiming to reveal its potential value and provide reference for its further development.

Preliminary study on Salvia miltiorrhiza bung endophytic fungus

Objective To select the strains which can produce tanshinone ⅡA like its host plant Salvia miltiorrhiza bung.Methods A total of 50 strains of endophytic fungi were isolated from healthy,living and symptomless tissues of Salvia miltiorrhiza bung,among which 29 strains were obtained from the root,14 from the stem,3 from the leaf,3 from the flower and 1 from the seed.Their antimicrobial activities against nine different bacteria,including both Gram-negative and Gram-positive bacteria,were measured by Oxford plate agar diffusion bioassay.Results Our data showed that all but four strains had significant antibacterial activities on at least one indicator bacterium to some extent,and five strains(DR1,DR4,DR16,DR18 and DF2)manifested quite prominent antibacterial activities against certain pathogenic bacteria.In some degree,it might indicate that this endophytic fungus isolated from the tissues of Salvia miltiorrhiza bung has a potential value as a natural antibacterial medicine as well.Thin layer chromatography(TLC)and high-performance liquid chromatography(HPLC)were carried out to test selected strains,both inside and outside of the cell to see if any strain can produce tanshinone ⅡA.The result showed that extracts from three strains,labeled as DR12(outside cell),DR21(inside cell)and DF3(inside cell),had a component with the same Rf value in TLC assay as that of authentic tanshinone ⅡA.The extract from DR12(outside cell)and DR21(inside cell)had a peak at retention time identical to that of authentic tanshinone ⅡA in HPLC.Conclusion The fungi appear to produce the bioactive compound tanshinone ⅡA,and they could be used to produce tanshinone ⅡA by fermentation.It provides a new way to synthesize this natural medicine.

Challenges and strategies in progress of drug delivery system for traditional Chinese medicine Salviae Miltiorrhizae Radix et Rhizoma(Danshen)

Traditional Chinese medicines(TCMs), with a history of thousands of years, are widely used clinically with effective treatment. However, the drug delivery systems(DDSs) for TCMs remains major challenges due to the characteristics of multi-components including alkaloids, flavones, anthraquinones, glycosides,proteins, volatile oils and other types. Therefore, the novel preparations and technology of modern pharmaceutics is introduced to improve TCM therapeutic effects due to instability and low bioavailability of active ingredients. Salviae Miltiorrhizae Radix et Rhizoma, the radix and rhizomes of Salvia miltiorrhiza Bunge(Danshen in Chinese), is a well known Chinese herbal medicine for protecting the cardiovascular system, with active ingredients mainly including lipophilic tanshinones and hydrophilic salvianolic acids.In this review, this drug is taken as an example to present challenges and strategies in progress of DDSs for TCMs. This review would also summary the characteristics of active ingredients in it including physicochemical properties and pharmacological effects. The purpose of this review is to provide inspirations and ideas for the DDSs designed from TCMs by summarizing the advances on DDSs for both singleand multi-component from Danshen.

Content of rare earth elements in Salvia miltiorrhiza bunge from different areas

Salvia miltiorrhiza bunge(SMB) has been widely used as a traditional Chinese medicine for a long time,and today more and more attention has been paid to the effective components in Chinese traditional medicine,especially to rare earth elements(REEs).14 kinds of REEs in SMB from different areas were analyzed by the methods of ICP-MS.The results showed that the total concentration of REEs(as REO,the same below) ranged from 8.43(as dry weight,the same below) to 37.30 mg/kg,and the concentrations of La,Ce and Nd were higher than 2 mg/kg except Nd in SMB from Beijing.So the total concentration of REEs in SMB was much higher than that in rice,corn and barley,which could be the mechanism of curative effect of SMB on cardiovascular cerebrovascular system,digestive system,respiratory system,countershock and antibiotic.The character of other elements and the content of REEs in soil from different areas should be responsible for the difference,but the allocation mechanism of REEs in SMB should be further studied.

Inhibition of Proliferation of Prostate Cancer Cell Line DU-145 in vitro and in vivo Using Salvia miltiorrhiza Bunge

Objective:To investigate the antiproliferative activity of Salvia miltiorrhiza Bunge.(SM)on the castration-resistant prostate cancer(CRPC)cell line DU-145 in vitro and in vivo.Methods:Prostate cancer cell line(DU-145)and normal prostate cell line(RWPE-1)were treated with SM at different concentrations(3.125,12.5,25 and 50μg/mL)to investigate the antiproliferative effects.DNA laddering analysis was performed to investigate the apoptosis of DU-145 cells.Molecular mechanism was investigated by Western blot analysis of p53,Bcl-2,prostate specific antigen(PSA),and androgen receptor(AR).Six-week-old male BALB/c nude mice were randomly divided into normal control group(n=101)and treated group(n=101)which administered 500 mg/kg SM for 2 weeks.Tumor volumes were measured.Results:Treatment with SM resulted in a dose-dependent decrease in cell number of DU-145 cells in comparison with RWPE-1.DNA laddering analysis indicated the apoptosis of DU-145 cells.Treatment with SM increased the expression of p53 and reduced the expression of Bcl-2 proteins.The levels of PSA were considerably reduced in SM-treated group compared to the controls,and a decrease in AR expression was observed when cells were treated with SM in the same pattern as a reduction in PSA.In the tumour xenograft study,SM given once a day for 2 weeks significantly inhibited tumour growth.Conclusion:SM might contribute to the anticancer actions such as induction of apoptosis and inhibition of proliferation of prostate cancer cells.

Electrochemical and spectroelectrochemical studies of four tanshinones from Salvia miltiorrhiza Bunge

The spectra of four tanshinones in the potentiostatic reduction process, including tanshinone I, dihydrotanshinone I, tanshinone IIA and cryptotanshinone, were investigated using spectroelectrochemical cell and UV spectrophotometer. Their cyclic voltammograms （CVs） were recorded with a glassy carbon electrode （GCE）. The experiment results show that the antioxidant activity of these tanshinones, in the structure, where A, B and C rings connect through a single double bond, is weaker than that where A ring does not have double bond. Moreover, the increasing angle strain in the reduction process could enhance the antioxidant activity. In summary, the rank of antioxidant activities of these tanshinones, from weak to strong, is tanshinone I, dihydrotanshinone I, tanshinone IIA and cryptotanshinone.

植物源性丹酚酸B对递增负荷训练大鼠骨骼肌铁死亡通路的影响

研究旨在探究植物源性丹酚酸B对递增负荷训练大鼠骨骼肌铁死亡通路的影响及其潜在的调控机制。本研究从草本植物丹参中提取分离得到纯化的丹酚酸B,按照实验设计喂食给大鼠,随后对大鼠进行递增负荷训练模拟了运动性疲劳状态,通过分析血清标志物和组织生化指标,研究了丹酚酸B在运动性疲劳过程中对氧化应激和铁代谢的影响。研究发现,丹酚酸B可剂量依赖性地减轻递增负荷训练引起的大鼠骨骼肌损伤,其中血清LDH和CK水平分别降低了50%和45%。丹酚酸B显著降低了骨骼肌中MDA水平,减轻了氧化应激。在铁代谢方面,丹酚酸B升高了骨骼肌中GSH水平,并上调了GPX4的转录和蛋白表达水平,抑制了铁死亡。丹酚酸B对铁代谢相关基因的转录和蛋白表达产生明显的影响,降低了Tf、TfR、DMT1的表达水平,提高了FTH1和FPN1的表达水平。结果显示,丹酚酸B通过调控铁代谢通路,参与抗运动性疲劳过程,为丹酚酸B作为天然抗氧化剂和铁死亡抑制剂的潜在机制提供了新见解。

bHLH transcription factor SmbHLH92 negatively regulates biosynthesis of phenolic acids and tanshinones in Salvia miltiorrhiza

Objective:Salvia miltiorrhiza is a valuable herbal medicine with tanshinone and phenolic acid as the main biological active ingredients.The biosynthetic regulation of these bioactive compounds is controlled by a set of transcription factors(TFs).The basic helix-loop-helix(bHLH)transcription factor plays an important role in various physiological and biochemical processes in plants.However,research on bHLH TFs regulating phenolic acid or tanshinone biosynthesis in S.miltiorrhiza is limited.Methods:qRT-PCR was used for gene expression analysis.The subcellular localization of SmbHLH92 was detected by SmbHLHg2-GFP transient transformation into tobacco leaves,and its fluorescence was observed using a confocal laser scanning microscope.The transcriptional activity of SmbHLH92 was confirmed in the AH109 yeast strain.RNA interference hairy roots of SmbHLH92-RNAi transgenic lines were obtained through Agrobacterium-mediated genetic transformation.Ultra performance liquid chromatography(UPLC)was used to detect the changes of phenolic acids and tanshinones.Results:SmbHLH92 is a bHLH transcription factor that is highly expressed in the root and phloem of S.miltiorrhiza.The subcellular localization and transcriptional activity of SmbHLH92 indicated that SmbHLH92 was located in the nucleus and may be a transcription factor.RNA interference(RNAi)of SmbHLH92 in hairy roots of S.miltiorrhiza significantly increased the accumulation of phenolic acid and tanshinone.Quantitative RT-PCR(RT-qPCR)analysis showed the transcription level of genes encoding the key enzymes involved in the phenolic acid and tanshinone biosynthetic pathways was increased in the hairy roots of the SmbHLH92-RNAi transgenic line,comparing with the control line.Conclusion:These data indicate that SmbHLH92 is a negative regulator involved in the regulation of phenolic acid and tanshinone biosynthesis in S.miltiorrhiza.

Transcriptional activity and subcellular location of SmWRKY42-like and its response to gibberellin and ethylene treatments in Salvia miltiorrhiza hairy roots

Objective: The exogenous gibberellin(GA) and ethylene(ET) treatment can improve the medicinal ingredients of Salvia miltiorrhiza. Interestingly, many reports pointed out that WRKY transcription factors played an important regulatory role in these treatment responses. However, whether the Sm WRKY mediate these treatment signalings in S. miltiorrhiza remains largely elusive.Methods: qRT-PCR was used for SmWRKY42-like in response to exogenous GA and ethephon(Eth) treatment. The subcellular location of SmWRKY42-like was transiently transformed into onion epidermal cells by particle bombardment. The self-activating activity of SmWRKY42-like was verified in AH109 yeast strain.Results: Sm WRKY42-like was a WRKY family gene in S. miltiorrhiza. The subcellular localization and transcriptional activity results of the SmWRKY42-like protein indicated that SmWRKY42-like mainly enriched in nucleus and might be a transcription factor in S. miltiorrhiza. In the meantime, the SmWRKY42-like gene significantly responded to exogenous GA and Eth treatment.Conclusion: These results collectively indicated the SmWRKY42-like gene functions, as an important hormone-responsive gene, might play a potentially role in ET and GA signaling pathways.

丹参活性成分丹酚酸B对运动性心肌损伤的保护作用及机制

本研究旨在探索从丹参中提取的最丰富的水溶性酚类化合物丹酚酸B在运动性心肌损伤中的保护作用以及其机制。结果显示,丹酚酸B剂量依赖性地改善了大鼠心功能,减轻了心肌损伤,并降低了血清中的心肌损伤标志物的水平。此外,丹酚酸B还抑制了心肌细胞凋亡和氧化应激反应,并调节了心肌组织中铁稳态。试验证实,丹酚酸B通过抑制铁死亡途径发挥了心肌保护作用。本研究结果表明,丹酚酸B可能成为一种天然药物,用于预防和治疗运动性心肌损伤。

丹参茉莉酸羧甲基转移酶的原核蛋白表达分析

茉莉酸羧甲基转移酶基因(jasmonic acid carboxyl methyltransferase,JMT)是植物茉莉酸甲酯(methyl jasmonate,MeJA)生物合成过程中的关键酶基因,在植物体内可依赖S-腺苷-蛋氨酸(S-adenosyladenylL-methionine,SAM)作为甲基供体,催化茉莉酸(jasmonic acid,JA)形成茉莉酸甲酯。本研究将丹参茉莉酸羧甲基转移酶基因cDNA构建到原核表达载体PET-28a(+)上后转化到大肠杆菌BL21(DE3)并加入异丙基-β-D-硫代半乳糖苷(IPTG)诱导表达。SDS-PAGE凝胶电泳检测结果表明该蛋白的分子量大小约44 kD,与预测的蛋白分子质量大小一致,从而证实丹参JMT基因在大肠杆菌中表达成功。丹参JMT基因在原核细胞表达将对此酶基因的蛋白纯化、体外酶活力检测以及基因功能等研究奠定一定的基础。