Study on Tanshinone Extracted from Salvia miltiorrhiza Bge. Dregs

[Objective]The aim was to extract tanshinone from Salvia miltiorrhiza Bge. dregs and to determine tanshinone components. [Method]Organic solvent method was adopted to extract tanshinone from S. miltiorrhiza dregs and TLC was used to determine the optimum extraction solvent. The components of tanshinone were measured with HPLC. [Result]Ether was the best solvent to extract tanshinone from S. miltiorrhiza dregs. After water immersion,dry dregs of S. miltiorrhiza and Panax notoginseng were extracted with ethanol to obtain fat-soluble extracts. Then with ether as the solvent for Soxhlet extraction,the yield of crude tanshinone was 2.17%. The HPLC detection showed that the contents of tanshinone Ⅱ A,methylene tanshinquinone,cryptotanshinone,tanshinone Ⅰ were 3.62%,1.02%,2.56%,2.75% respectively. [Conclusion]The components of tanshinone in S. miltiorrhiza dregs were basically the same as tanshinone in medicine S. miltiorrhiza. S. miltiorrhiza dregs could be used as a kind of tanshinone resource,which has the value of development and utilization.

Effects of Salvia miltiorrhiza Bge.f.alba on neuronal regeneration following cerebral ischemia/reperfusion

BACKGROUND： Subsequent to cerebral ischemic injury, endogenous neural stem cells are activated, but ischemia-induced neuronal loss is not compensated by ischemic injury-induced neural regeneration. Salvia （S.） miltiorrhiza Bge.f.alba （Baihua Danshen, a Chinese herbal medicine） could enhance learning and memory functions, as well as promote neural regeneration. OBJECTIVE： To observe the effects of S. miltiorrhiza Bge.f.alba on recovery from cerebral ischemia-reperfusion injury, and the influence on neuronal regeneration and differentiation. DESIGN, TIME AND SETTING： Randomized, controlled, animal experiments were performed at the Experimental Animal Center and Neurobiology Laboratory of Taishan Medical College in September of 2006. MATERIALS： S. miltiorrhiza Bge.f.alba was provided by Taishan Medical College Botanic Garden, Taian, China; dl-3n-butylphthalide （NBP） soft capsule was purchased from NBP Pharmaceutical, Shijiazhuang, China; mouse anti-bromodeoxyuridine antibody, rabbit anti-NF200 antibody, and bromodeoxyuridine were purchased from Sigma, Louis, MO, USA; Annexin V-fluorescein isothiocyanate/PI apoptosis kit was purchased from Nanjing Comissariado Biological Technology Development, Nanjing, China. METHODS： Adult Sprague Dawley rats were randomly assigned to sham surgery, model （cerebral ischemia and reperfusion, without administration）, S. miltiorrhiza Bge.f.alba, and NBP groups. Following establishment of the cerebral ischemia/reperfusion model, S. miltiorrhiza Bge.f.alba or NBP （1 mL/100 g） was respectively perfused at 30 minutes following cerebral ischemia/reperfusion. MAIN OUTCOME MEASURES： Alterations in cerebral blood flow before and after ischemia/reperfusion, NF200- and bromodeoxyuridine-double positive cells in striatum of affected tissues, as well as neuronal apoptosis rate at days 5 and 7 following cerebral ischemia/reperfusion. RESULTS： Subsequent to cerebral ischemia reperfusion, cerebral blood flow was reduced. Following treatment with S. miltiorrhiza Bge.f.alba, cerebral blood flow significantly increased （P 〈 0.05）. NBP treatment was inferior to S. miltiorrhiza Bge.f.alba with regard to stabilization of cerebral blood flow （P 〈 0.05）. S. miltiorrhiza Bge.f.alba significantly increased the number of newly formed neurons in rats following cerebral ischemia （P 〈 0.05） and significantly reduced neuronal apoptosis （P 〈 0.05）, with no significant difference compared with NBP treatment （P 〉 0.05）. CONCLUSION： S. miltiorrhiza Bge.f.alba significantly increased cerebral blood flow, reduced neuronal apoptosis, and promoted neuronal regeneration in rats with cerebral ischemia/reperfusion impairment.

Visualizing the spatial distribution and alteration of metabolites in continuously cropped Salvia miltiorrhiza Bge using MALDI-MSI

Salvia miltiorrhiza Bge(SMB)has long been used in traditional Chinese medicine to treat cardiovascular and cerebrovascular diseases.Growing clinical usage has led to a huge demand for artificial planting of SMB.Thus,continuous cropping of SMB is an important challenge that needs to be addressed.Continuous cropping can alter the metabolic profile of plants,resulting in poor growth and low yield.In this study,we tried to image the spatial location and variation of endogenous metabolites in continuously cropped SMB using matrix-assisted laser desorption/ionization mass spectrometry imaging(MALDIMSI).Spatially resolved expressions of tanshinones,salvianolic acids,polyamines,phenolic acids,amino acids,and oligosaccharides in normal and continuously cropped SMB roots were compared.The expressions of dihydrotanshinone Ⅰ,tanshinone Ⅱ A,dehydromiltirone,miltirone,dehydrotanshinone ⅡA,spermine,salvianolic acid B/E,tetrasaccharide,and pentasaccharide in continuously cropped SMB roots were much lower than those in normal roots.There was little difference in the expressions of caffeic acid and salvianolic acid A in normal and continuously cropped SMB roots.Ferulic acid was more widely distributed in xylem of normal SMB but strongly expressed in xylem,phloem,and cambium of continuously cropped SMB.The spatially resolved metabolite information enhances our understanding of the metabolic signature of continuously cropped SMB and also provides insights into the metabolic effects of continuous cropping in other plants.

Effect of Salvia Miltiorrhiza Bge on Left Ventricular Hypertrophy and the Expression of Tumor Necrosis Factor-α in Spontaneously Hypertensive Rats

The effects of salvia miltiorrhiza Bge (SMB) on left ventricular hypertrophy (LVH) and the expression of tumor necrosis factor-α (TNF-α) in the left ventricle of spontaneously hypertensive rats and the action mechanism were investigated. Normal Wistar-kyoto (WKY) rats were used as negative control, and spontaneously hypertensive rats (SHR) were randomly assigned to receive pla- cebo or SMB. SMB (1 g/kg·d) was injected intraperitoneally for 12 weeks. Systolic blood pressure (SBP) and left ventricular mass index (LVMI) were measured. HE, VG and immunohistochemical staining combined with computed morphometry were employed to evaluate the cardiomyocyte size, diameter, the collagen volume fraction (CVF), perivascular circumferential area (PVCA), and tumor necrosis factor-α (TNF-α) expression in the left ventricular tissue. The results showed, as compared with WKY rats, the SBP, LVMI, cardiomyocyte size, diameter, CVF, PCVA, and TNF-α expression were increased markedly in the 20-week-old spontaneously hypertensive rats. SMB decreased LVMI (P<0.01), size of cardiomyocytes (P<0.01), collagen volume fraction (P<0.01), perivascular circum- ferential area (P<0.01), and TNF-α expression (P<0.01), but had no effect on SBP (P>0.05). It was suggested that chronic administration of SMB could inhibit and reverse the development of LVH in spontaneously hypertensive rats independent of BP. TNF-α may be involved in the reversal mecha- nism of LVH by SMB.

Salvia miltiorrhiza extract may exert an anti-obesity effect in rats with high-fat diet-induced obesity by modulating gut microbiome and lipid metabolism

BACKGROUND Studies have shown that a high-fat diet(HFD) can alter gut microbiota(GM)homeostasis and participate in lipid metabolism disorders associated with obesity.Therefore, regulating the construction of GM with the balance of lipid metabolism has become essential for treating obesity. Salvia miltiorrhiza extract(Sal), a common traditional Chinese medicine, has been proven effective against atherosclerosis, hyperlipidemia, obesity, and other dyslipidemia-related diseases.AIM To investigate the anti-obesity effects of Sal in rats with HFD-induced obesity, and explore the underlying mechanism by focusing on GM and lipid metabolism.METHODS Obesity was induced in rats with an HFD for 7 wk, and Sal(0.675 g/1.35 g/2.70g/kg/d) was administered to treat obese rats for 8 wk. The therapeutic effect was evaluated by body weight, body fat index, waistline, and serum lipid level. Lipid factors(cAMP, PKA, and HSL) in liver and fat homogenates were analyzed by ELISA. The effect of Sal on GM and lipid metabolism was assessed by 16S rRNAbased microbiota analysis and untargeted lipidomic analysis(LC-MS/MS),respectively.RESULTS Sal treatment markedly reduced weight, body fat index, serum triglycerides(TG), total cholesterol(TC), low-density lipoprotein, glucose, free fatty acid, hepatic lipid accumulation, and adipocyte vacuolation, and increased serum high-density lipoprotein(HDL-C) in rats with HFD-induced obesity. These effects were associated with increased concentrations of lipid factors such as c AMP, PKA, and HSL in the liver and adipose tissues, enhanced gut integrity, and improved lipid metabolism. GM analysis revealed that Sal could reverse HFD-induced dysbacteriosis by promoting the abundance of Actinobacteriota and Proteobacteria, and decreasing the growth of Firmicutes and Desulfobacterita. Furthermore, LC-MS/MS analysis indicated that Sal decreased TGs(TG18:2/18:2/20:4, TG16:0/18:2/22:6), DGs(DG14:0/22:6, DG22:6/22:6), CL(18:2/18:1/18:1/20:0), and increased ceramides(Cers;Cer d16:0/21:0, Cer d16:1/24:1),(O-acyl)-ω-hydroxy fatty acids(OAHFAs;OAHFA18:0/14:0) in the feces of rats. Spearman’s correlation analysis further indicated that TGs, DGs, and CL were negatively related to the abundance of Facklamia and Dubosiella, and positively correlated with Blautia and Quinella, while OAHFAs and Cers were the opposite.CONCLUSION Sal has an anti-obesity effect by regulating the GM and lipid metabolism.

Determination of inhibitory activity of Salvia miltiorrhiza extracts on xanthine oxidase with a paper-based analytical device

A novel paper-based analytical device(PAD)was prepared and applied to determine the xanthine oxidase(XOD)inhibitory activity of Salvia miltiorrhiza extracts(SME).First,polycaprolactone was 3D printed on filter paper and heated to form hydrophobic barriers.Then the modified paper was cut according to the specific design.Necessary reagents including XOD for the colorimetric assay were immobilized on two separate pieces of paper.By simply adding phosphate buffer,the reaction was performed on the double-layer PAD.Quantitative results were obtained by analyzing the color intensity with the specialized device system(consisting of a smartphone,a detection box and sandwich plates).The 3Dprinted detection box was small,with a size of 9.0 cm×7.0 cm×11.5 cm.Color component G performed well in terms of linearity and detection limits and thus was identified as the index.The reaction conditions were optimized using a definitive screening design.Moreover,a 10%glycerol solution was found to be a suitable stabilizer.When the stabilizer was added,the activity of XOD could be maintained for at least 15 days under 4℃ or-20℃ storage conditions.The inhibitory activity of SME was investigated and compared to that of allopurinol.The results obtained with the PAD showed agreement with those obtained with the microplate method.In conclusion,the proposed PAD method is simple,accurate and has a potential for point-of-care testing.It also holds promise for use in rapid quality testing of medicinal herbs,intermediate products,and preparations of traditional Chinese medicines.

Molecular Identification of Salvia miltiorrhiza Bge. and Its Adulterants Based on ITS2 Sequences

[ Objectives] The study was conducted to investigate the molecular identification of Salvia miltiorrhiza Bge. and its adulterants by DNA barcoding andspecific primer PCR. [ Methods] With ITS2 sequenceas DNA barcode, the materials were amplified by PCR and sequenced, and the NJ phylogenetic tree was constructed. The secondary structure of ITS2 was predicted by database and its website established by Koetschan et al. , and the self-designed primers were used to carry out specific primer PCR identification. [Results] ITS2 sequence length was around 470 bp. The results of cluster analysis showed that S. miltiorrhiza Bge. and its adulterants were clustered on different branches and showed monophyly. The comparison of secondary structure showed that S. miltiorrhiza Bge. had little differences from S. przewalskii, while there were significant differences from A. lappa in the number, size and location of stem-loop and the rotation angle of the spiral arm from the central ring. The specific primers could distinguish the S. miltiorrhiza Bge. and its counterfeits by PCR technique. [Conclusions] DNA barcoding and specific primer PCR are effective in distinguishing S. miltiorrhiza Bge. and its adulterants, and it has an important application foreground in the identification of Chinese herbal medicines.

Effects of salvia miltiorrhiza bge on myocardial fibrosis in hypertensive rats

Objective: To study the effects of Salvia Miltiorrhiza Bge (SMB) on myocardial fibrosis in hypertension in rats. Methods: Normal Wistar kyoto rats served as negative control (group A) . The rats with hypertension were divided into 2 groups: the hypertension group (group B) which served as the positive control and the SMB-treated hypertension group (group C). The effects of SMB on the systolic pressure, the myocardial content of aldosterone (ALD) and nitric oxide (NO) and the activity of superoxide dismutase (SOD) in the myocardium were detected. Results: The systolic pressure of the rats of group C showed no marked change after the administration of SMB and showed no difference from that of the rats of group B. The content of collagen Ⅰ and Ⅲ of the left ventricle, the collagen Ⅰ /collagen Ⅲ ratio and the content-of ALD were lower in group C than in group B and higher in group C than in group A. The content of NO and the activity of SOD in the myocardium were higher in group C than in group B and lower in group C than in group A. Conclusion: It was concluded that SMB was able to prevent or reverse the progress of myocardial fibrosis resulting from hypertension. The mechanism of this effect of SMB might be due to the fact that SMB can decrease the collagen synthesis and ALD content and increase collagen decomposition, NO content and SOD activity in the myocardium.

Protective effect and possible mechanisms of Salvia miltiorrhiza Bge. for the treatment of diabetic nephropathy: A systematic review and meta-analysis of animal studies

Objective:To explore the protective effect and potential mechanisms of danshen root(Salvia miltiorrhiza Bge.,S.miltiorrhiza) and its extracts for the treatment of diabetic nephropathy(DN).Methods:Preclinical studies of S.miltiorrhiza and its extracts on DN were systematically searched in nine databases.The primary outcomes were blood glucose,kidney function,proteinuria,and renal histopathology.The secondary outcomes included the related mechanisms.The methodological quality of animal studies was assessed based on the risk of bias tool of the Systematic Review Centre for Laboratory Animal Experimentation(SYRCLE) for animal studies.Meta-analysis was performed using R software(version 4.1.2).Results:Twenty-nine animal experimental studies that met the eligibility criteria were included in this study.Compared to the control group,S.miltiorrhiza reduced the serum creatinine,blood urea nitrogen,24-h urine protein,24-h urine albumin,blood glucose,and kidney index(kidney weight/body weight),and alleviated renal pathological damage.In terms of the mechanism of action,compared to the control group,S.miltiorrhiza reduced the levels of transforming growth factor β1,collagen Ⅳ,malondialdehyde,tumor necrosis factor α,interleukin-6,and monocyte/macrophage(ED-1),and increased the levels of superoxide dismutase,glutathione peroxidase,nuclear factor E2-related factor 2,and heme oxygenase-1.Conclusion:The existing evidence shows that S.miltiorrhiza has beneficial effects on the animal model of DN,and its mechanism is mainly related to improving kidney fibrosis,oxidative stress,and inflammatory response.

Compound Astragalus and Salvia miltiorrhiza extract exerts anti-tumor activity by intervening the interaction of microRNA-145/microRNA-21 and Smad3C/3L phos-phorylation in hepatocellular carcinoma

OBJECTIVBE To investigate the intervention of compound Astragalus and Salvia miltiorrhiza extract(CASE) consisted of astragalosides,astragalus polysaccharides and salvianolic acids on the interaction of microRNA-145/microRNA-21(miR-145/miR-21) and Smad3 C/3 L phosphorylation(pSmad3 C/pSmad3 L) down-stream of transforming growth factor-β(TGF-β)/mitogen activated protein kinase(MAPK) signaling in hepatocellular carcinoma(HCC) progression by in vitro and in vivo experi.ments.METHODS In HepG2 cells and xenografts of nude mice,antagomir/agomir and plasmids of Smad3 C/3 L phosphorylation site mutation(Smad3 3 S-A/Smad3 EPSM) were used to intervene miR-145/miR-21 and pSmad3 C/pSmad3 L expression respectively,then incorporative CASE treatment.Cell proliferation,migration,apoptosis,tumor growth and histopathologic characteristics of xenografts,relevant proteins of TGF-β/Smad pathway and miR-145/miR-21 were evaluated.RESULTS CASE up-regulated miR-145 while down-regulated miR-21,inhibited cell proliferation,migration and tumor growth,accelerated cell apoptosis in HepG2 cells respectively transfected with Smad3 WT,Smad3 EPSM,Smad3 3 S-A plasmids in cultured dishes and xenografts of nude mice,the above effects were more evident in HepG2 cells with increased pSmad3 C.In TGF-β1-stimulated HepG2 cells and xenografts of nude mice,CASE antagonized the facilitating effects of miR-145 antagomir/miR-21 agomir on cell migration,proliferation,tumor growth and inhibiting effects of miR-145 antagomir/miR-21 agomir on cell apoptosis;CASE increased miR-145 down-regulated by miR-145 antagomir and decreased miR-21 up-regulated by miR-21 agomir,reduced protein level of pSmad3 L and their proteins including TβRⅡ,pERK1/2,pJNK1/2 and pp38 while elevated pSmad3 C expression.CONCLUSION These results suggest that pSmad3 C/pSmad3 L maybe interact with miR-145/miR-21 in HCC progression,which may be one of important molecular mechanisms of CASE's anti-HCC effects.

Tyrosine Aminotransferase Gene (SmTAT) Revealed Genetic Diversity and Phylogeny of Cultivated Danshen (Salvia miltiorrhiza) Populations

Chinese traditional medicine Danshen is the radix of the perennial herbs of Salvia miltiorrhiza Bunge, which has a variety of pharmacological effects and is traditionally and extensively applied clinically to treat cardiovascular disorders. In this research, the genomic genes for tyrosine aminotransferase (TAT) of 38 cultivated populations of Danshen in China were cloned and bioinformatic analyses were conducted to reveal its genetic diversity and phylogeny. The full-length SmTAT was 2296 - 2444 bp including 6 exons (encoding 411 amino acids) and 5 introns. Overall, the SmTAT genes in cultivated Danshen populations are highly conserved with a relative low level of genetic diversity. The spliced exons (1236 bp) had 23 SNP variations with a rate of 1.86%, of which 22 occurred in the white flower S. miltiorrhiza Bge.f.alba population (W-SCHY-W-1) and led to 5 amino acid variations. The entire 290 SNP variations with a rate of 24% in the 5 introns occurred exclusively in W-SCHY-W-1. Phylogenetic trees based on the full-length, combined introns, the spliced exons, and the deduced amino acid sequences of SmTAT all showed a two-clade basic structure with W-SCHY-W-1 uniquely standing alone. The SmTAT gene of the white flower population (W-SCHY-W-1) is unique and especially rich in variations. The first time clarified genomic SmTAT gene structure and genetic diversity in cultivated Danshen populations laid an excellent foundation for further studies on the biosynthesis of bioactives and the molecular breeding of Danshen as well as in plant tyrosine metabolism.

不同垄作方式和种植密度对丹参产量与主要成分含量的影响

为探究不同栽培方式和生长时期对丹参根与地上部产量和主要活性成分含量的影响,本研究以紫花丹参为试验材料,采用小垄单行[三种密度:H1(8万株·hm^(-2))、H2(10万株·hm^(-2))、H3(13.3万株·hm^(-2))]与大垄双行[三种密度:H4(8万株·hm^(-2))、H5(10万株·hm^(-2))、H6(13.3万株·hm^(-2))]两种垄作方式,动态取样,测定并分析不同垄作方式、种植密度和生长时期的单株重、产量、主要活性成分含量及积累规律。结果表明,大垄双行丹参单株干重、产量、根的主要活性成分含量平均值均高于小垄单行。根的单株干重和产量的最佳处理分别为H5(48.09 g)、H4(5986.69 kg·hm^(-2)),均在11月19日最高;地上部单株干重和产量的最佳处理均为H4(50.34 g、6712.75 kg·hm^(-2)),均在10月14日最高。根中主要活性成分总含量(总丹参酮+总黄酮+总酚酸)在两种垄作方式下均呈“升-降-升”的变化趋势,最佳处理为H4(295.4 mg·g^(-1)),在9月7日最高;叶、茎中(总黄酮+总酚酸)总含量的最佳处理分别为H4(238.42 mg·g^(-1))、H6(122.43 mg·g^(-1)),均在10月14日最高;丹参酮ⅡA、丹参酮I、隐丹参酮和丹酚酸B四种活性成分总含量的最佳处理为H6(63.23 mg·g^(-1))。综上,大垄双行栽培方式优于小垄单行。本研究结果为丹参的合理种植及根与地上部综合开发利用提供了理论依据。

胆汁酸诱导胃癌的发生机制及丹参提取物潜在作用的研究进展

胆汁反流可导致胃黏膜发生炎性反应、癌变。胆汁酸可以通过激活各种受体和信号通路来诱导胃癌的发生。传统中药丹参提取物可以起到预防胆汁酸诱导胃癌的作用。文章对近年来胆汁酸在胃癌发生中的作用机制进行分析,胃黏膜长时间暴露于胆汁酸中会导致黏膜损伤及炎性反应,导致慢性胃炎、肠化生、异型增生和胃癌;丹参提取物在预防和治疗胆汁酸诱导的胃癌中具有潜在优势,丹参素通过调节多种涉及炎症、氧化应激、DNA损伤和细胞凋亡的信号通路,可以预防胆汁反流引起胃炎和胃癌。

丹参提取物联合瑞芬太尼通过调控Nrf2信号通路减轻缺氧复氧诱导的心肌细胞损伤

为探讨丹参提取物联合瑞芬太尼在减轻缺氧/复氧诱导的心肌细胞损伤中的作用及其机制。本研究将H9c2心肌细胞分为对照组、模型组、丹参提取物组(Tan I组)、瑞芬太尼组(RPC)组和联合组。对照组细胞不做处理,模型组缺氧/复氧(HR)诱导的心肌细胞损伤模型,Tan I组在模型组的基础上给与1μmol/LTan I孵育24h,RPC组在模型组的基础上给与1μmol/LRPC孵育24h联合组在模型组的基础上同时给与1μmol/L Tan I和1μmol/L RPC孵育24 h。采用CCK-8测定各组细胞活力流式细胞术测定各组细胞凋亡率,采用Western blot测定各组细胞Nrf2和HO-1表达水平进一步转染Nrf2-siRNA分析Nrf2信号通路对Tan I联合RPC保护HR诱导H9c2心肌细胞损伤的影响。结果表明,HR诱导后H9c2心肌细胞的活力显著下降,药物处理后,Tan I组、RPC组和联合组H9c2细胞的细胞活力显著增加,且联合组的细胞活力改善与Tan I组和RPC组相比更显著;HR诱导后H9c2心肌细胞的凋亡率显著升高,药物处理后,Tan I组、RPC组和联合组H9c2细胞的细胞凋亡率显著下降,且联合组的细胞凋亡率下降水平与Tan I组和RPC组相比更显著;HR引起H9c2细胞Nrf2总蛋白表达和核Nrf2蛋白表达水平显著降低,各组给药后增加了总Nrf2蛋白和核Nrf2蛋白的表达,降低了细胞质中的Nrf2蛋白,且与Tan I组和RPC组相比联合组总Nrf2蛋白和核Nrf2蛋白的表达水平显著增高,细胞质中的Nrf2蛋白表达水平显著降低;转染Nrf2-siRNA后,Tan I联合RPC处理的H9c2心肌细胞中Nrf2和HO-1的表达均下降。丹参提取物联合瑞芬太尼可通过调控Nrf2信号通路减轻HR诱导的心肌细胞损伤。

丹参酚酸类化合物的合成生物学研究进展

丹参酚酸类化合物是丹参发挥保护心血管系统、抗肿瘤与抗纤维化等多种药理作用的一类关键物质基础。以合成丹酚酸A、丹酚酸B和迷迭香酸等丹参酚酸类化合物为目标的微生物细胞工厂的构建,可以实现丹参酚酸类化合物异源高效合成,缓解临床对丹参药材的用药压力。本文就丹参酚酸类化合物的合成生物学、生物合成关键催化酶及其代谢调控等研究现状进行了归纳分析,以期为丹参酚酸类化合物的高效合成与资源应用提供科学依据。

基于网络药理学探讨丹参-葛根药对治疗心肌纤维化的作用机制

目的采用网络药理学探讨丹参-葛根药对治疗心肌纤维化的活性成分及作用机制。方法从中药系统药理学分析平台获取丹参-葛根药对活性成分和作用靶点,通过GeneCards数据库获取心肌纤维化的相关靶点,使用Venny 2.1软件获取两者共同靶点;运用STRING数据库和Cytoscape 3.7.1软件构建共同靶点的蛋白-蛋白互作(PPI)网络并进行拓扑学分析;采用ClusterProfiler R功能包对共同靶点进行基因本体(GO)功能和KEGG通路富集分析;最后使用Cytoscape 3.7.1软件构建“活性成分-靶点-通路”网络并分析。结果筛选得到丹参-葛根药对候选活性成分30个,活性成分和心肌纤维化共同靶点41个。共同靶点PPI网络的平均点度值为19.7,平均介数为19.1,度值和介数均超过平均值的靶点共有14个。KEGG显著富集到73条通路,其中与心肌纤维化相关的通路有6条。“活性成分-靶点-通路”网络显示,丹参中的木犀草素、丹参酮IIA和葛根中的葛根素、β-谷甾醇等活性成分通过共同调控脂质与动脉粥样硬化、糖尿病并发症中的AGE-RAGE、血流剪切力与动脉粥样硬化、缺氧诱导因子-1(HIF-1)、肿瘤坏死因子(TNF)、白细胞介数-17(IL-17)等信号通路起到抗心肌纤维化的功效。结论揭示了丹参-葛根药对多成分、多靶点、多通路治疗心肌纤维化的作用特点,为进一步研究丹参-葛根药对治疗心肌纤维化的作用机制提供理论依据和新的思路。

Rheum palmatum L. and Salvia miltiorrhiza Bge. Alleviates Acute Pancreatitis by Regulating Th17 Cell Differentiation: An Integrated Network Pharmacology Analysis, Molecular Dynamics Simulation and Experimental Validation

Objective: To identify the core targets of Rheum palmatum L. and Salvia miltiorrhiza Bge.,(Dahuang-Danshen, DH-DS) and the mechanism underlying its therapeutic efficacy in acute pancreatitis(AP)using a network pharmacology approach and validate the findings in animal experiments. Methods: Network pharmacology analysis was used to elucidate the mechanisms underlying the therapeutic effects of DH-DS in AP. The reliability of the results was verified by molecular docking simulation and molecular dynamics simulation.Finally, the results of network pharmacology enrichment analysis were verified by immunohistochemistry,Western blot analysis and real-time quantitative PCR, respectively. Results: Sixty-seven common targets of DH-DS in AP were identified and mitogen-activated protein kinase 3(MAPK3), Janus kinase 2(JAK2), signal transducer and activator of transcription 3(STAT3), protein c-Fos(FOS) were identified as core targets in the protein interaction(PPI) network analysis. Gene ontology analysis showed that cellular response to organic substance was the main functions of DH-DS in AP, and Kyoto Encyclopedia of Genes and Genomes analysis showed that the main pathway included Th17 cell differentiation. Molecular docking simulation confirmed that DH-DS binds with strong affinity to MAPK3, STAT3 and FOS. Molecular dynamics simulation revealed that FOS-isotanshinone Ⅱ and STAT3-dan-shexinkum d had good binding capacity. Animal experiments indicated that compared with the AP model group, DH-DS treatment effectively alleviated AP by inhibiting the expression of interleukin-1β, interleukin-6 and tumor necrosis factor-α, and blocking the activation of Th17 cell differentiation(P<0.01). Conclusion: DH-DS could inhibit the expression of inflammatory factors and protect pancreatic tissues,which would be functioned by regulating Th17 cell differentiation-related m RNA and protein expressions.

丹参-葛根不同配伍比例对体外抗氧化活性的影响

为探究不同配伍比例的丹参与葛根体外抗氧化能力,采用水提法提取丹参葛根13个不同配伍比例(分别为1∶0、5∶1、3∶1、2∶1、3∶2、4∶3、1∶1、3∶4、2∶3、1∶2、1∶3、1∶5、0∶1)的有效成分,利用紫外分光光度法测定丹参葛根水提物的DPPH及ABTS自由基清除能力,以评估不同配伍比例的丹参葛根抗氧化活性。结果表明:丹参较葛根具有更强的抗氧化能力;与丹参、葛根单味药相比,丹参与葛根配伍比例为1∶2、3∶1和4∶3时,其水提物表现出更强的清除DPPH及ABTS自由基能力。综上,丹参与葛根配伍使用能增强其体外抗氧化活性,可为丹参葛根配伍应用提供一定的科学依据。

丹参-三七药对治疗冠心病的药效和机制研究进展

近些年来,丹参和三七在临床上常被用来治疗冠心病如心肌缺血、动脉粥样硬化,以及与冠心病等危的糖尿病和高血压等。因此,丹参-三七药对及其组分配伍治疗冠心病的潜在机制受到广泛关注。大量文献表明丹参-三七及其组分配伍在治疗冠心病中具有调节脂质代谢和能量代谢、抗炎、抗血小板聚集、改善内皮细胞功能作用。总结了丹参-三七药对及其组分配伍在治疗冠心病中的药效和机制,以期为其后续研究提供更全面的理论依据。

基于离子液体原位泡腾萃取法在丹参及西洋参克百威残留分析中的应用

本研究建立了一种新颖的基于酸性离子液体的原位泡腾萃取技术的前处理方法。酸性离子液体1-丁基-3-甲基咪唑硫酸氢盐呈酸性,能与碳酸氢盐反应产生二氧化碳,起到泡腾萃取的效果,它还能与六氟磷酸铵发生离子交换作用而置换成疏水性离子液体1-丁基-3-甲基咪唑六氟磷酸盐,使其与水相分离,通过离心获取萃取相,最后通过高效液相色谱仪测定萃取相中农药的含量。考察了酸性离子液体、碳酸氢钠和氯化钠用量对萃取效率的影响,最终所得最佳条件为200 mg酸性离子液体、0.4 g碳酸氢钠和1.0 g氯化钠。在最优条件下,克百威的线性范围为0.3~4.0μg/mL,日内和日间的相对偏差(RSD)为4.6%和10.2%,丹参和西洋参在4.0μg/mL加标水平下,加标回收率为94.7%(西洋参)和74.6%(丹参),将此方法可用于不同基质中药材和饮片中农药残留的处理。