The Effect of Soil Enzymes and Polysaccharides Secreted by the Roots of Salvia miltiorrhiza Bunge under Drought,High Temperature,and Nitrogen and Phosphorus Deficits

Root exudates serve as crucial mediators for information exchange between plants and soil,and are an important evolutionary mechanism for plants’adaptation to environmental changes.In this study,15 different abiotic stress models were established using various stress factors,including drought(D),high temperature(T),nitrogen deficiency(N),phosphorus deficiency(P),and their combinations.We investigated their effects on the seedling growth of Salvia miltiorrhiza Bunge and the activities of Solid-Urease(S-UE),Solid-Nitrite Reductase(S-NiR),Solid-Nitrate Reductase(S-NR),Solid-Phosphotransferase(S-PT),and Solid-Catalase(S-CAT),as well as the contents of polysaccharides in the culture medium.The results showed that the growth of S.miltiorrhiza was inhibited under 15 stress conditions.Among them,13 stress conditions increased the root-shoot ratio.These 15 stress conditions significantly reduced the activity of S-NR,two combinations significantly improved the activity of S-NIR,they were synergistic stresses of high temperature and nitrogen deficiency(TN),and synergistic stresses of drought and nitrogen deficiency(DN)(p<0.05).The activity of S-UE was significantly improved under N,D,T,synergistic stresses of drought and high temperature(DT),DN,synergistic stresses of drought and phosphorus deficiency(DP),and synergistic stresses of high temperature,nitrogen,and phosphorus deficiency(TNP)stress conditions(p<0.05).Most stress combinations reduced the activity of S-PT,but D and T significantly improved it.(p<0.05).The N,DN,and TN stress conditions significantly reduced S-CAT activity.The P,DT,and synergistic stresses of drought,high temperature,and phosphorus deficiency(DTP)significantly decreased the total polysaccharide content of the soil(p<0.05).The research suggested that abiotic stress hindered the growth of S.miltiorrhiza and altered the behavior of root secretion.Roots regulated the secretion of several substances in response to various abiotic stresses,including soil nitrogen cycle enzymes,phosphorus transport-related enzymes,and antioxidant enzymes.In conclusion,plants regulate the utilization of rhizosphere substances in response to abiotic stresses by modulating the exudation of soil enzymes and polysaccharides by the root system.At the same time,soil carbon sequestration was affected by the adverse environment,which restricted the input of organic matter into the soil.

Study on Optimization of Ethanol Reflux Extraction of Phenolic Acids from Salvia miltiorrhiza

The extraction technology of phenolic acid compounds from Salvia miltiorrhiza by ethanol reflux was studied. In this experiment, salvianolic acid B standard was used to make the standard curve. Single factor experiment and orthogonal experiment were used to study the extraction of different ethanol concentrations, reflux times and material-to-liquid ratios. The OD value of salvianolic acid compounds was measured with a spectrophotometer. The extraction rate of phenolic acid compounds under different extraction conditions was calculated through a regression equation, so as to obtain the optimal conditions for the ethanol reflux extraction process of Salvia miltiorrhiza. The experimental data can provide a reference for the ethanol reflux extraction process of salvianolic acids in the industry. According to the experiment, the extraction rate of phenolic acids in Salvia miltiorrhiza was the highest when the ethanol concentration was 60%, the reflux time was 1.5 hours, and the ratio of material-to-liquid was 1:10.

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.

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.

Investigation on Genetic Characteristics of Salvia miltiorrhiza Bge. by Using cpSSR Technology

[Objective] This study aimed to analyze the genetic characteristics of Salvia miltiorrhiza by using cpSSR （chloroplast microsatellites） molecular marker method. [Method] Twelve pairs of SSR primers with ideal cpSSR amplification result, good repeatability and clear amplification bands were selected for cpSSR detection and analysis of S. miltiorrhiza samples from 31 sampling locations in 25 counties of 8 provinces in China. [Result] S. miltiorrhiza shows an overall medium level in the cytoplasmic inheritance （cpSSR） and varying degrees of differences in the regions. Based on Shannon’s Information index （I） and Nei’s genetic diversity index, the cytoplasmic genetic diversity in different provinces shows a decreasing order of Shanxi, Henan, Shandong, Hebei, Sichuan, Jiangsu, Shaanxi, Anhui. Genetic variations of S. miltiorrhiza in eight provinces were found mainly among populations. Gene flow among populations with in a province is less than that among different provinces. [Conclusion] Comprehensive analysis shows that S. miltiorrhiza cultivated in genuine producing areas and traditional main producing areas are mainly introduced from the local before introduction of some foreign germplasm resources during cultivation. The exchange of S. miltiorrhiza germplasm resources exists very early among genuine producing areas such as Sichuan, Shandong and Henan, while S. miltiorrhiza germplasm resources in new producing areas are mainly introduced from genuine producing areas. However, no geographical correlation was observed in the genetic differentiation, which further reveals that there is a wide range of genetic exchange among S. miltiorrhiza germplasm resources in China, resulting from the artificial introduction from allopatry.

Microwave digestion polarography for determining seven trace elements in Salvia Miltiorrhiza Root and compound Salvia Miltiorrhiza Root injection simultaneously

The sensitive second derivative waves of Cu(Ⅱ),Pb(Ⅱ),Cd(Ⅱ),Ni(Ⅱ),Zn(Ⅱ),Fe(Ⅱ)and Mn(Ⅱ),forming in the substrate solution(pH=9.26)consisting of ammonia,ammonium chlorid,gelatin and ascorbic acid were researched.The peak potentials of the polarographic waves of the seven ions are at about 0.48,0.63,0.79,1.04,1.28,1.44 and 1.60 V respectively.Combining with microwave technique,a new method for the simultaneous determination of the seven trace elements in Chinese traditional medicine was developed.The method is easy to operate,rapid,simple and convenient.When the signal-to-noise rate equals 3,the detection limits of Cu,Pb,Cd,Ni,Zn,Fe and Mn are 4.2×10 3,5.3×10 3,2.1×10 3,5.8×10 4,3.0×10 3,7.7×10 4 and 1.1×10 3 μg/mL respectively.Well linear relationships exist between the concentrations and the peak currents when Cu,Pb,Cd,Ni,Zn,Fe and Mn concentrations are within 8.5×10 3 10,9.7×10 3 10,4.5×10 3 10,1.2×10 3 10,6.4×10 3 10,1.5×10 3 10 and 2.8×10 3 10 μg/mL,respectively.The method has been used to the simultaneous determination of the seven trace elements in Salvia Miltiorrhiza Root and compound Salvia Miltiorrhiza Root injection,the relative standard deviations(RSDs)of the Cu,Pb,Cd,Ni,Zn,Fe and Mn in the two medicines are 3.9% and 5.8%,4.0% and 4.1%,4.3% and 5.7%,4.9% and 5.3%,4.4% and 4.7%,3.5% and 4.0%,0.51% and 2.8%,respectively;the comparisons of the determination results with the values obtained by the standard method indicate that the presented method has very well veracity.

Dynamic of Root Dry Weight of Salvia miltiorrhiza Bunge

ObjectiveThis study was to investigate the dynamic of root dry weight of Salvia miltiorrhiza to provide a basis for optimal management in field cultivation. MethodThe Salvia miltiorrhiza Bunge seedlings were transplanted at three different dates November 26, 2009 （T 1 ）, March 9, 2010（T 2 ） and March 27, 2010（T 3 ）, and at three different densities 20 cm × 25 cm （D 1 ）, 25 cm × 25 cm （D 2 ）, 25 cm × 30 cm （D 3 ）. So a total of nine treatments were finally set. The samples were collected regularly for measuring the root dry weight and plotting the dynamic curves. Normalized root weight was fitted with CURVE EXPERT 1.3. ResultLogistic equation was the most superior equation, and estimated respectively two inflection points and length of fast growing time of S. miltiorrhiza. ConclusionThe fast-growing time of S. miltiorrhiza root was in 1 900 ℃·d to 2 200 ℃·d GDD.

Preliminary Study on Induction and Identification of Polyploidy of Shandong Salvia miltiorrhiza Buds Treated by Colchicine

[Objective] The aim was to study the feasibility of using colchicine to induce polyploidy from the buds of Shandong Salvia miltiorrhiza.[Method] The buds of Shandong Salvia miltiorrhiza were induced by colchicine.[Result] When the concentration of colchicine supplemented in culture medium was 12 mg/L,the induction rate of polyploidy was the highest（36.6%）,but its survival rate was only 16.7%.When the concentration of colchicine was 8 mg/L,the induction rate of polyploidy was 15.9%,and its survival rate was up to 40.0%.Polyploidy plant of Shandong Salvia miltiorrhiza was strong and had thick,large and dark leaves,and long stomata on the lower epidermis of leaves had small density.[Conclusion] Colchicine could induce buds effectively,and the polyploidy breeding material of Salvia miltiorrhiza was obtained,which laid a foundation for shortening breeding process.

山东丹参(Salvia miltiorrhiza)不同地理居群的遗传多样性

以山东泰安、临沂、莱芜、菏泽和潍坊5个居群的72份丹参株系为材料,利用ISSR引物进行群体遗传结构的研究。结果表明:8个ISSR引物在5个居群中共扩增出219个位点,平均可扩增出27条带,在种级水平及泰安、临沂、莱芜、菏泽和潍坊5个居群水平多态性位点百分比分别为98.63%、81.28%、66.67%、66.21%、51.14%和50.68%,种级水平的Nei基因多样性和Shannon信息指数大于各居群;5个居群Nei基因多样性和Shannon信息指数相比较,泰安>临沂>莱芜>菏泽>潍坊;根据基因分化系数,测得的基因流值Nm为4.2352;UPGMA聚类分析结果表明,莱芜居群和临沂居群遗传一致度最大,遗传关系最近,泰安居群与其它4个居群遗传关系最远;分析发现菏泽居群、泰安居群是相对独立的群体,但5个居群间存在部分基因交流。所有参数分析表明,泰安居群遗传多样性最丰富,故在制定原位种质保护计划时应优先考虑泰山周边地区的丹参。

Gastric mucosal injury due to hemorrhagic reperfusion and efficacy of Salvia miltiorrhizae extract F and cimetidine

AIM: To observe the gastric mucosal injury caused by hemorrhagic shock and reperfusion and to compare the effect between Salvia miltiorrhizae extract F (SEF) and cimetidine (CI) on it. METHODS: A model of hemorrhage/reperfusion injury was produced by Itoh method. Wistar rats were randomly divided into three groups: 0.9% sodium chloride treatment group (NS group), SEF treatment group (SEF group), and CI treatment group (CI group). Saline, SEF and CI were injected respectively. The index of gastric mucosal lesions (IGML) was expressed as the percentage of lesion area in the gastric mucosa. The degree of gastric mucosal lesions was categorized into grades 0, 1, 2, 3. Atom absorption method was used to measure the intracellular calcium content. Radioimmunoassay was used to measure the concentrations of prostaglandins. RESULTS: IGML (%) and grade 3 (%) were 23.18±6.82, 58.44±9.07 in NS group, 4.42±1.39, 20.32±6.95 in SEF group and 3.74±1.56, 23.12±5.09 in CI group, and the above parameters in SEF group and CI group decreased significantly (IGML: SEF vs NS, t=6.712, P=0.000<0.01; CI vs NS, t=6.943, P=0.000<0.01; grade 3: SEF vs HS, t=8.386, P=0.000; CI vs HS, t=8.411, P= 0.000), but the grade 0 and grade 1 damage in SEF group (22.05±5.96, 34.12±8.12) and CI group (18.54±4.82, 30.15±7.12) were markedly higher than those in NS group (3.01±1.01, 8.35±1.95; grade 0: SEF vs HS, t=8.434, P=0.000<0.01; CI vs NS, t=7.950, P=0.000<0.01; grade 1: SEF vs NS, t =8.422, P=0.000<0.01; CI vs NS, t=8.448, P=0.000<0.01). The intracellular calcium content (μg/mg) in SEF group (0.104±0.015) and CI group (0.102±0.010) was markedly lower than that in NS group (0.131±0.019, SEF vs NS, t=2.463, P=0.038<0.05; CI vs HS, t=3.056, P=0.017<0.05). The levels (pg/mg) of PGE_2, 6-keto-PGF_(1α) and 6-keto-PGF_(1α)/TXB_2 were 540±183, 714±124,17.38±5.93 in NS group and 581±168, 737±102, 19.04±8.03 in CI group, 760±192,1 248±158, 33.42±9.24 in SEF group, and the above parameters in SEF group markedly raised (PGE_2: SEF vs NS, t=2.282, P=0.046<0.05; SEF vs CI, t=2.265, P=0.047<0.05; 6-keto-PGF_(1α): SEF vs NS, t=6.583, P=0.000<0.000; SEF vs CI, t=6.708, P=0.000<0.01; 6-keto-PGF_(1α)/TXB_2: SEF vs NS, t=3.963, P=0.003<0.001; SEF vs Cl, t=3.243, P=0.009<0.01), whereas TXB_2 level in SEF group (45.37±7.54) was obviously lower than that in NS group (58.28±6.74, t=3.086, P=0.014<0.05) and CI group (54.32±6.89, t=2.265, P=0.047<0.05). No significant difference was shown between NS group and CI group (PGE_2: t=0.414, P=0.688>0.05; 6-keto-PGF_(1α): t=0.310, P=0.763>0.05; TXB_2: t=1.099, P=0.298>0.05; 6-keto-PGF_(1α)/TXB_2: t=0.372, P=0.718>0.05). CONCLUSION: Both SEF and CI could inhibit reperfusioninduced injury in gastric mucosa, but with different mechanisms. SEF could not only enhance the protective effect of gastric mucosa, but also abate the injury factors, while CI can only abate the injury factors.

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.

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.

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.

Effects of purified herbal extract of Salvia miltiorrhiza on lipid profile in hyperlipidemic patients

Background and Objective Hyperlipidemia is one of the most potent and best substantiated risk factors for coronary heart disease （CHD）. Purified Salvia miltiorrhiza extract （PSME） had been shown to have hypolipidemic effect in animal experiments. This study aimed to evaluate its lipids modulating effects in hyperlipidemic patients. Methods In this single-blind, placebo cantrolled study, lipid profiles of 80 hyperlipidemic patients were checked at same conditions. They were divided into two equal groups randomly （each composing of 40 patients）. They were given PSME tablet （800 mg） three times per day, or placebo tablet. All patients were put on NCEP type II diet and six weeks later, lipid profiles were checked. Results In PSME group, total cholesterol decreased by 27.32 mg/dl （12.3% reduction）, LDL-cholesterol decreased by 23.13 mg/dl （16.8% reduction） and HDL-cholesterol increased by 9.06 mg/dl （11.1%）, all were statistically significant. Although triglyceride dropped by 12.12 mg/dl （5.1%） but this was not significant statistically （P=0.34）. There were no significant changes oflipids levels in the placebo group. Conclusions PSME has significant favorable effect on total cholesterol, LDL-cholesterol, and HDL-cholesterol and may be a potential agent for the treatment ofatherogenic dyslipidemia.

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.

Effect of Salvia Miltiorrhiza on Left Ventricular Hypertrophy and Cardiac Aldosterone in Spontaneously Hypertensive Rats

Chronic treatment with Salvia Miltiorrhiza preventing left ventricular hypertrophy (L VH) and its possible mechanism- inhibiting the action of cardiac aldosterone in spontaneously hypertensive rats (SHR) were investigated.Normotensive Wistar- kyoto (WKY ) rats and SHRs were used.Part of SHRs was treated with Salvia Miltiorrhiza for 12 weeks.Systolic blood pres- sure (SBP) and left ventricular mass index were measured.Sections of heart tissue were stained with HE method and Van Gieson method.Collagen volume fraction was determined in the leftven- tricle by automatically quantitative m orphometry.Cardiac aldosterone concentration was measured by radioimm unoassay.The results indicated thatcom pared with WKY rats,SHRs exhibited high- er SBP,left ventricular collagen volume fraction,and aldosterone concentration (all P<0 .0 5 ) . After the treatm ent with Salvia Miltiorrhiza,SBP,left ventricular collagen volum e fraction,and aldosterone concentration in SHR were decreased as compared with control group (P<0 .0 5 ) ex- cept SBP.It was concluded thatchronic treatment with Salvia Miltiorrhiza could preventleftven- tricular hypertrophy in SHR,significantly inhibit collagen compositions in left ventricle.The m echanism was probably related with the inhibition of the cardiac aldosterone action.

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.

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.