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 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.

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.

Experimental Study on the Preventive Mechanism of Salviae Miltiorrhizae Against Atherosclerosis in Rabbits Models

Summary: The preventive mechanism of salviae miltiorrhizae (SM) against experimental atherosclerosis (AS) in rabbits models was investigated. The experimental AS rabbit models were reproduced by feeding the high cholesterol diet. The changes of atherosclerotic plaques in normal group, model group and SM treated group were observed. The levels of serum TG, TC, HDL-C and LDL-C were determined. The immunohistochemistry was used to detect the expression of Bcl-2, Bax and IL-6 proteins in atherosclerotic plaques. The results showed that the level of serum TG in SM treated group was significantly lower than in model group (P<0.01). Immunohistochemistry revealed that the expression of Bcl-2, Bax and IL-6 in model group was significantly higher than in normal group. In the SM group, the expression of Bcl-2 protein was up-regulated and that of Bax was down-regulated. It was suggested that SM could inhibit formation of AS in experimental rabbits. To decrease the expression of Bax and increase the expression of Bcl-2 protein may be one of the mechanisms of SM against atherosclerosis.

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.

Clinical study of adjuvant therapy on ischemic stroke with salviae miltiorrhizae and ligustrazine

Objective: To study the effect of adjuvant therapy with salviae miltiorrhizae and ligustrazine injection on neural function injury, degree of oxidative stress reaction, chemotactic factor and adhesion molecule in patients with ischemic stroke. <br> Methods: Patients with ischemic stroke admitted in our hospital from August 2013 to October 2015 were retrospectively analyzed and divided into conventional treatment (CT) group receiving conventional treatment and traditional Chinese medicine auxiliary (TCMA) group receiving adjuvant therapy with salviae miltiorrhizae and ligustrazine injection. After the treatment of the first month and the third month, serum was collected to detect the contents of nerve injury molecules, oxidative stress parameters, chemotactic factor and adhesion molecule. <br> Results: After the 4-week treatment, the contents of serum neuron specific enolase, S100 calcium-binding protein B, heart-type fatty acid binding protein, malondialdehyde, advanced oxidation protein products, 8-hydroxy-2'-deoxyguanosine, monocyte chemoattractant protein-1,CD40,CD40 ligand, vascular endothelial cadherin, soluble intercellular adhesion molecule 1 and soluble vascular adhesion molecule 1 in patients ofTCMA group were all significantly lower than those of CT group, and the contents of superoxidase dismutase, glutathione peroxidase and catalase were significantly higher than those of CT group. After the 8-week treatment, the contents of serum neuron specific enolase, S100 calcium-binding protein B, heart-type fatty acid binding protein, malondialdehyde, advanced oxidation protein products, 8-hydroxy-2'-deoxyguanosine, monocyte chemoattractant protein-1,CD40, CD40 ligand, vascular endothelial cadherin, soluble intercellular adhesion molecule 1 and soluble vascular adhesion molecule 1 in patients ofTCMA group were all significantly lower than those of CT group and the contents of superoxidase dismutase, glutathione peroxidase and catalase were significantly higher than those of CT group. <br> Conclusions: Adjuvant therapy with salviae miltiorrhizae and ligustrazine injection can alleviate the neural function injury, inhibit oxidative stress reaction and the generation of chemotactic factor and adhesion molecule in patients with ischemic stroke, which is an effective medicine for treating ischemic stroke.

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.

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.

Hypoglycemic Effect of Radix Et Rhizoma Salviae Miltiorrhizae,Guixi Green Tea and Their Compatibility on Mice

[Objectives]To explore the hypoglycemic effect of Radix Et Rhizoma Salviae Miltiorrhizae,Guixi green tea and their compatibility on mice,so as to provide more options for the adjuvant treatment of diabetes and improve the comprehensive utilization value of Guixi green tea.[Methods]After being fed adaptively for 7 d,60 SPF-grade male mice were intraperitoneally injected with alloxouracil(180 mg/kg)to induce hyperglycemia model,from which 35 mice with a successful model were selected.They were randomly divided into 5 groups(Danshen group,green tea group,compatible group,positive control group and blank control group),with 7 animals in each group.The mice in all groups were administered according to the dose of 200 mg/kg once a day for 15 d.After the last administration,their body weight was measured on the 15 th day after fasting for 12 h,and their fasting blood glucose was measured by tail clipping.[Results]After modeling,the mice in each group showed typical hyperglycemia symptoms,namely obvious polydipsia,polyphagia,polyuria and weight loss.Compared with the blank control group,Danshen group and green tea group had obvious hypoglycemic effect(P<0.05),and compatible group had significant hypoglycemic effect(P<0.01).Compared with the blank control group,the weight loss of diabetic mice could also be inhibited in green tea group and compatible group(P<0.05).[Conclusions]Danshen,Guixi green tea and their compatibility had the effect of lowering blood sugar,among which the compatible group had the most obvious effect.

Effects of Lycii Fructus and Salviae Miltiorrhizae on the Syndrome of Deficiency with Blood Stasis in RCS(rdy-/-, p-/-) Rats with Retinitis Pigmentosa: An Intervention Study

Objective To investigate the effects of Lycii Fructus(LF,Gou Qi Zi,枸杞子)and Salviae Miltiorrhizae Radix Ex Rhizoma(SM,Dan Shen,丹参)on the syndrome of deficiency with blood stasis in the RCS(rdy-/-,p-/-)rats with retinitis pigmentosa(RP).Methods A total of 32 RCS(rdy-/-,p-/-)rats were divided into 4 groups(equal amounts of female and male rats in each group):model group treated with 0.9%normal saline,LF group treated with LF formula granules,SM group treated with SM formula granules,and LF and SM(L·S)group treated with LF and SM formula granules.Eight RCS(rdy+/+,p+/+)rats(4 males and 4 females)were treated with 0.9%normal saline to serve as blank group.The contents of E2,PG,P-Selectin,plasma viscosity,whole blood relative index of the high shear rate and fibrinogen content in plasma,and the content of cAMP and cGMP in retinal homogenate were detected.The retina was evaluated by hematoxylin-eosin staining.Results The contents of E2,PG,P-Selectin,plasma viscosity,whole blood relative index of the high shear rate,and fibrinogen content in the plasma of L·S group significantly differed from those of model group(P<0.01),but were similar to those of blank group.The contents of cAMP and cGMP in the retinal homogenate of L·S group significantly differed from those in model group(P<0.01)but were similar to those in blank group(P>0.05).Conclusions LF and SM can effectively treat retinitis pigmentosa by ameliorating the syndrome of deficiency with blood stasis.

EFFECTS OF LIMONENE,SALVIA MILTIORRHIZA AND TURMERIC DERIVATIVES ON H-RAS ONCOGENE EXPRESSION AND GAP JUNCTION INTERCELLULAR COMMUNICATION IN HUMAN SOLID TUMOR CELL LINES

Objective: To study gap junction intercellular communication (GJIC), H ras oncogene expression and ras oncogene product (P 21 ras protein) expression in four human solid tumor cell lines, W1-38,CACO 2,A549 and PaCa, and the effects of four compounds, Salvia miltiorrhiza derivative (SMD), d Limonene, Turmeric derivative I (TD I) and Turmeric derivative II (TD II), on them. Methods: The abilities of the four solid tumor cell lines to transfer dye to adjacent cells were examined by the scrape loading/dye transfer technique, and the H ras oncogene expression by Northern blotting and P 21 ras protein expression by Western blotting. Results: The results showed the loss of intercellular coupling in PaCa cells, slight GJIC in A549 and CACO 2 cells, and a good GJIC in W1-38 cells. The four compounds could improve the GJIC of PaCa to different extents. The amount of total and membrane associated P 21 ras in PaCa cells were decreased after treatment with SMD, d Limonene and TD I (2.5 μg/ml) for 48 h. Concomitantly, the growth of PaCa cells decreased in soft agar and had enhanced GJIC. The relative potency was found to be:d Limonene>SMD >TD I=TD II. There was no significant effect of the four compounds on H ras oncogene expression. Conclusion: It was suggested that there was an excellent correlation between loss of Lucifer Yellow dye transfer and ras gene mutation rate in the four solid tumor cell lines (ras gene mutation rate inversely correlated with average cell number coupled, r=0.98) i.e., the high ras gene mutation was closely correlated with loss of GJIC in these malignant human tumor cells; The antitumor effect of the monoterpene d Limonene and the phenol compound, SMD, might be related to inhibition of P 21 ras membrane association and enhancement of GJIC, whilst that of the others may be by a different mechanism; The inhibition of P 21 ras membrane association was directly related to the enhancement of gap junction intercellular com munication.

Effect of Salvia miltiorrhiza Injection Combined with Laparoscopic Reduction on Injury of Appendix in Patients with Intussusception

[Objectives] The aim was to analyze the effect of Salvia miltiorrhiza injection combined with laparoscopic reduction on the degree of appendix injury in intussusception.[Methods]Seventy-four children with intussusception who were treated in the Xuchang Municipal Hospital from August 2014 to August 2017 were randomly and evenly divided into two groups,control group and treatment group.The children in the control group were treated by laparoscopic reduction,and those in the treatment group were treated with S.miltiorrhiza injection combined with laparoscopic reduction.The clinical effect on intussusception in the children was compared between the two groups.[Results] Before treatment,no significant difference was founding serum NO level between the control and treatment groups(P > 0.05).On Day 1,3 and 5 of treatment,the serum NO levels of the treatment group[(74.16 ± 6.81),(61.98 ± 4.77) and(54.76 ± 4.36) μmol/L] were significantly lower than those of the control group[(89.67 ± 6.34),(84.75 ± 5.21) and(71.95 ± 4.58) μmol/L](P < 0.05).After treatment,the TNF-α and IL-6 levels of the treatment group[(184.32 ± 10.67) and(332.16 ± 34.17) pg/m L] were significantly lower than those of the control group[(268.26 ± 12.74) and(438.23 ± 35.26) pg/m L](P < 0.05);and the Chiu score of the treatment group(2.34 ± 1.12) was also significantly lower than that of the control group(4.85 ± 1.67)(P < 0.05).[Conclusions]For children with intussusception,S.miltiorrhiza injection combined with laparoscopic reduction is effective in reducing serum NO level and reducing the degree of appendix injury,which is worthy of clinical application.

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.

The Inhibitory Effect of Radix Salviae Miltiorrhizae on Hypoxic Structural Remodeling of Intra－acinarPulmonary Arteries

The inhibitory effect of Radix Salviae Miltiorrhizae (RSM) on hypoxic structural remodeling of intra-acinar pulmonary arteries (IAPA) was observed by light and electron microscopy and morphometry. It was found that RSM can not only dilate IAPA and relieve the hypoxic injuries to endothelia cells,but also inhibit the active muscularization of IAPA in the hypoxic animals, suggesting that RSM plays a very important role in inhibiting structural remodeling of IAPA and pulmonary hypertension.

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.

EFFECT OF RADIX SALVIAE MILTIORRHIZAE ON GROWTH OF ISOLATED CELLS FROM EMBRYONIC CHICKEN FRONTAL BONE CULTURED IN VITRO (A HISTOCHEMICAL STUDY) Ⅱ.THE DEVELOPMENT AND MATURATION OF OSTEOBLAST-LIKE CELLS

The isolated osteoblast-like cells from embryonic chicken frontal bone werecultured in vitro and histochemical methods adopted to observe the effect of RadixSalviac Miltiorrhizae (RSM) on proliferation, differentiation, and osteogenic capacity ofthese cells. It was found that: 1. The mitosis and proliferation of the osteoblast-like cellscould be accelerated by RSM, resulting in increased density of the cells in RSM groupas compared with the control. 2. After 48 h, the pseudopodia stretched out and drew backactively in osteoblast-like cells in RSM group. Small particles produced in the cells weresecreted through exocytosis to the extracellular medium. However, in the control group,the capacity to form and secrete these particles was limited. These particles showed posi-tive Alcian blue staining in Alcian blue-Sirius red reaction, so they were acidmucopolysaccharide particles. 3. The osteoblast-like cells could secrete vesicular particles 3micra in diameter. These vesicular particles could be stained with Alcian blue in earlystage, then they could be stained with Sirius red, and finally by Alizarin red S. Thesevesicular particles could aggregate and fuse around the cell colonies, forming bonenodules and bone flakes. The quantity and volume of the bone nodules and flakes inRSM group were larger than in the control group. 4. The bone nodules and flakes couldbe labeled vitally with tetracycline, and show strong yellow fluorescence under thefluorescence microscope. Therefore, these substances were the newly formed bone sub-stances.

EFFECTS OF SALVIA MILTIORRHIZA BUNGE(SMB)ON LIPID PEROXIDATION AND PLASMA MEMBRANE FLUIDITY OF CULTURED HUMAN FETAL HEPATOCYTES INJURED BY CARBON TETRACHLORIDE

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.

Effects of Salvia Miltiorrhiza Bung on the formation of osteoclastlike cells in mouse bone marrow culture

Objective:To studytheeffectsof SalviaMiltiorrhizaBung(SMB)on theformationof osteoclast-likecellsin vitro.Methods:Mousebonemarrowcellswereculturedwithα-MEMandexposedto SMB(1.0g/L),PGE 2 (1×10 -7 mol/L)or1.25(OH) 2 D 3 (1×10 -8 mol/L)only,andSMB+PGE 2 ,SMB+1.25(OH) 2 D 3 for8d respectively.Tatrate-resistant acidphosphatase(TRACP)-positivecellsweredetectedwithBurstonemethod.Results:Thenumberof TRACPpositive cells(mononuclearcellsonly)ingroupof SMBwas29.8%of thatof thecontrol,thatingroupof combinationof SMB withPGE 2 was35.2%of thatof groupPGE 2 alone,andthatingroupof SMBplus1.25(OH) 2 D 3 was39.3%of thatof group1.25(OH) 2 D 3 alone.SMBinhibitedtheTRACP-positivecellsformationonlyat theconcentrationof1.0g/L,whichwere notonlyinmononuclearcellsbutalsoinmultinuclearcells.In addition,TRACP-positivecellsinthegroupsof1.25(OH) 2 D 3 or PGE 2 were6.68and6.04timesrespectivelyas thatof thecontrols.Conclu sion:SMBhastheinhibitingeffectson osteoclast-likecellsformation,bothosteoclastsandtheirprecursors.

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.

A comparative study of Salvia miltiorrhiza Radix & Rhizoma raw material and granule products using chromatographic analysis and antioxidant activity

Objective: Granules of herbal extracts are a popular medicinal preparation consumed in traditional Chinese medicine clinical practice. However, their quality and efficacy evaluation are lacking. This study aimed to compare the quality and anti-oxidant activity of Dan Shen(Salvia miltiorrhiza Radix & Rhizoma)granule extracts with their herbal extracts.Methods: Chromatographic method was used to determine the content of 7 marker compounds in the water extracts of the herb compared to that of 12 granule extracts. Agglomerative hierarchical clustering(AHC) and principal component analysis(PCA) distinguished the herbal and granule extracts based on the content of the marker compounds. The antioxidant activities of herbal and granule extracts were evaluated by 2, 2'-azino-bis(3-ethylbenz-thiazoline-6-sulfonic acid)(ABTS), organic chemical compound 2,2-diphenyl-1-picrylhydrazyl(DPPH) and ferric ion reducing antioxidant power(FRAP) assays.Results: The herbal extracts group showed significantly higher contents of salvianolic acid B, sodium danshensu and cryptotanshinone compared with that of the granule group. This corresponded to significantly higher ABTS, DPPH and FRAP(P <.05) activities of the herbal extracts. The AHC and PCA analysis distinguished granule extracts from most herbal extracts predominantly by the content of salvianolic acid B.Conclusion: The results confirm the need for the assessment of granule products so that healthcare practitioners and consumers are better informed of their quality and efficacy.