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Objective::To screen out the effective components of Salvia miltiorrhiza by establishing an in vitro model of pulmonary epithelial mesenchymal transformation. Method::Different concentrations of salvianolic acid A (10, 20, 40, 80, 160 μmol·L-1), salvianolic acid B (10, 20, 40, 80, 160 μmol·L-1), tanshinol (10, 20, 40, 80, 160 μmol·L-1), tanshinoneⅡA (10, 20, 40, 80, 160 μmol·L-1) and the blank group were applied to A549 cell, cell proliferation and cytotoxicity assay (MTS) were used to detect the proliferation effect of menthol on A549 cells.After screening the safe concentration of the active ingredients of salvia miltiorrhiza by MTS, cells were divided into blank group, model group, salvianolic acid A group, salvianolic acid B group, tanshinol group and tanshinoneⅡA.Then, the inhibitory effect of the active ingredients of salvia miltiorrhiza on the proliferation of A549 cells induced by TGF-β1 was detected by MTS. Enzyme linked immunosorbent assay (ELISA) method to detect salvia miltiorrhiza effective component of fiber protein(FN), collagen type I (COL-Ⅰ) expression. Based on the above results, the active components of salvia miltiorrhiza, which have best inhibition were screened out, and their effects on the expression of E-calcium-viscosity (E-Cad) protein were detected by Western blot. Result::Compared with blank group, salvianolic acid A 40 μmol·L-1, salvianolic acid B 160 μmol·L-1, tanshinol 160 μmol·L-1 had toxic effects on A549 cells (P<0.05). In the non-toxic concentration range, compared with the model group, salvianolic acid A 10, 20 μmol·L-1, salvianolic acid B 80 μmol·L-1 showed inhibition effect after 24 h culture (P<0.05). After 72 h culture, salvianolic acid A 5, 10, 20 μmol·L-1, salvianolic acid B 40, 80 μmol·L-1inhibition effect was very significant (P<0.01). ELISA results showed that with the blank group, model group cells the expression of FN and COL-Ⅰ increased significantly (P < 0.01). Compare with model group, salvianolic acid A 20 μmol·L-1, salvianolic acid B 80 μmol·L-1 inhibited FN and COL-Ⅰ(P<0.05). Western blot results showed that salicylic acid A and salicylic acid B had protective effects on E-Cad (P<0.01). Conclusion::Salvianolic acid A and salvianolic acid B have inhibitory effects on epithelial mesenchymal transformation by TGF-β1, which may be the main effective components of salvianolic acid in the treatment of pulmonary fibrosis.
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OBJECTIVE:To establish a method for simultaneous determination of tanshinol,caffeic acid,rosmarinic,salviano-lic acid B,salvianolic acid A,tanshinoneⅠ,cryptotanshinone,tanshinone ⅡA and ursolic acid in Compound xueshuantong cap-sules. METHODS:UPLC-MS/MS method was adopted. The determination was performed on ACQUITY UPLC? BEH C18 column with mobile phase consisted of acetonitrile-0.1%formic acid(gradient elution)at the flow rate of 0.2 mL/min. The column tempera-ture was 40 ℃,and the temperature of injector was 10 ℃. Analysis time was 7 min,and sample size was 5 μL. The electrospray ionization source(ESI)was used;ion source temperature was 150℃;capillary voltage was 3.5 kV;cone flow was 50 L/h;desol-vation temperature was 350 ℃;desolvation gas flow was 650 L/h;nebuliser pressure was 7 × 105 Pa;ion monitoring and multiple reaction monitoring (MRM) was performed. RESULTS:The linear ranges of tanshinol,caffeic acid,rosmarinic,salvianolic acid B,salvianolic acid A,tanshinoneⅠ,cryptotanshinone,tanshinone ⅡA and ursolic acid were 10.0-100.0 μg/mL (r=0.9998), 0.1-1.0 μg/mL(r=0.9998),4.0-40.0 μg/mL(r=0.9999),10.0-100.0 μg/mL(r=0.9999),15.0-150.0 μg/mL(r=0.9997), 8.0-80.0 μg/mL(r=0.9998),10.0-100.0 μg/mL(r=0.9997),50.0-500.0 μg/mL(r=0.9997)and 6.0-60.0 μg/mL(r=0.9998), respectively. The limits of quantitation were 40.0,9.6,38.0,88.0,130.0,39.0,4.4,3.2 and 10.0 ng/mL,separately. The limits of detection were 12.0,3.0,11.0,26.0,39.0,12.0,1.3,1.0 and 3.0 ng/mL,respectively. RSDs of precision,stability and repro-ducibility tests were all lower than 3%. The recoveries were 97.34%-103.20%(RSD=2.19%,n=6),97.22%-102.39%(RSD=2.03%,n=6),98.51%-101.70%(RSD=1.32%,n=6),97.86%-102.49%(RSD=2.09%,n=6),96.75%-103.12%(RSD=2.36%,n=6),98.43%-101.65%(RSD=1.25%,n=6), 97.59%-101.50%(RSD=1.50%,n=6), 96.45%-102.88%(RSD=2.58%,n=6),97.02%-103.11%(RSD=2.38%,n=6),separately. CONCLUSIONS:The method is simple and accurate,and can be used for simultaneous determination of 9 components in Compound xueshuantong capsules.
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Objective@#To investigate the protective effect and potential mechanism of tanshinol borneol ester (TBE) on homocysteine(Hcy) induced rat bone marrow mesenchymal stem cells (BMSCs) damage.@*Methods@#BMSCs were isolated and cultured in vitro by density gradient centrifugation and adherent culture method. BMSCs were divided into the control (normal isolation and culture), TBE-1(10 μmol/L TBE-1 solution with 100 μl), TBE-2 (10 μmol/L TBE-2 solution with 100 μl), Hcy (0.5 mmol/L Hcy solution with 100 μl), Hcy + TBE-1(0.5 mmol/L Hcy solution with 100 μl, and 10 μmol/L TBE-1 solution with 100 μl), Hcy + TBE-2 (0.5 mmol/L Hcy solution with 100 μl, and 10 μmol/L TBE-2 solution with 100 μl), Hcy+ TBE-1+ inhibitor group(0.5 mmol/L Hcy solution with 100 μl, 10 μmol/L TBE-1 solution with 100 μl, and 25 μmol/L LY294002(specific blocker of phosphatidylinositol 3 kinase) solution with 100 μl), Hcy+ TBE-2+ inhibitor group(0.5 mmol/L Hcy solution with 100 μl, 10 μmol/L TBE-2 solution with 100 μl, and 25 μmol/L LY294002 solution with 100 μl). Cell proliferation activity was detected by MTT assay. The T-SOD activity and malonaldehyde level of cells were measured by anthineoxidase method and TBA method, respectively, to evaluate cell oxidative and antioxidative activities. The ultrastructure of cells was observed under transmission electron microscope. The expression level of PKB and NF-κB of cells in various groups were detected with the immunocytochemical method.@*Results@#(1)Cell proliferation activity in TBE-1 group and TBE-2 group was significantly increased compared with the control group (both P<0.01), and was similar between TBE-1 group and TBE-2 groups after 1, 12, 24 and 48 hours treatment.(2)The T-SOD activity in TBE-1 group and TBE-2 group was significantly higher than in control group (both P<0.01), while it was significantly lower in Hcy group, Hcy+ TBE-1 group, and Hcy+ TBE-2 group than in control group(all P<0.01), and was similar between control group, Hcy+ TBE-1+ inhibitor group, and Hcy+ TBE-2+ inhibitor group(all P>0.05). The T-SOD activity was higher in Hcy+ TBE-1 group and Hcy+ TBE-2 group than in Hcy group(both P<0.01), and was higher in Hcy+ TBE-1+ inhibitor group than in Hcy+ TBE-1 group(P<0.05) and was higher in Hcy+ TBE-2+ inhibitor group than in Hcy+ TBE-2 group(P<0.05). The malonaldehyde level was lower in TBE-1 group and TBE-2 group than in control group(both P<0.01), was higher in Hcy group, Hcy+ TBE-1 group, Hcy+ TBE-2 group, Hcy+ TBE-1+ inhibitor group, and Hcy+ TBE-2+ inhibitor group than in control group(all P<0.01), was lower in Hcy+ TBE-1 group and Hcy+ TBE-2 group than in Hcy group(both P<0.01), was higher in Hcy+ TBE-1+ inhibitor group than in Hcy+ TBE-1 group(P<0.05), was higher in Hcy+ TBE-2+ inhibitor group than in Hcy+ TBE-2 group(P<0.05). (3)Under electron microscope, BMSCs showed profound swelling, senescence and apoptosis of cells increased significantly in Hcy group, Hcy+ TBE-1+ inhibitor group, and Hcy+ TBE-2+ inhibitor group when compared with control group. BMSCs in the TBE-1 and TBE-2 groups presented with abundant rough endoplasmic reticulum, and very active cell metabolism signs. Compared with Hcy group, BMSCs edema, the number of aging and apoptotic cells, and cell injury severity were significantly less in TBE-1+ Hcy group and TBE-2+ Hcy. (4)The PKB level was higher in TBE-1 group and TBE-2 group than in control group(both P<0.01), was lower in Hcy group, Hcy+ TBE-2 group, Hcy+ TBE-1+ inhibitor group, and Hcy+ TBE-2+ inhibitor group than in control group(all P<0.01), was similar between control group and Hcy+ TBE-1 group(P>0.05), was higher in Hcy+ TBE-1 group and Hcy+ TBE-2 group than in Hcy group(both P<0.05), was lower in Hcy+ TBE-1+ inhibitor group than in Hcy+ TBE-1 group(P<0.05), and was lower in Hcy+ TBE-2+ inhibitor group than in Hcy+ TBE-2 group(P<0.05). The NF-κB level was higher in TBE-1 group and TBE-2 group than in control group(both P<0.01), was lower in Hcy group, Hcy+ TBE-1 group, Hcy+ TBE-2 group, Hcy+ TBE-1+ inhibitor group, and Hcy+ TBE-2+ inhibitor group than in control group(P<0.01 or 0.05), was higher in Hcy+ TBE-1 group and Hcy+ TBE-2 group than in Hcy group(both P<0.05), was lower in Hcy+ TBE-1+ inhibitor group than in Hcy+ TBE-1 group(P<0.05) and was lower in Hcy+ TBE-2+ inhibitor group than in Hcy+ TBE-2 group(P<0.05).@*Conclusion@#Tanshinol borneol ester can promote the proliferation of BMSC, and attenuate the homocysteine induced rat BMSCs damage possibly through activation of phosphatidylinositol 3 kinase/PKB signal transduction and its downstream signal pathway protein NF-κB.
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Objective To study the chemical constituents of Mallotus furetianus. Methods The constituents were isolated and purified by silica gel chromatography and HPLC repeatedly, and the structures were identified by spectra analysis and chemical methods. Results Sixteen compounds were isolated from the leaves of M. furetianus, the structures were identified as 3-hydroxy-4,5 (R)-dimethy1-2 (5H)-furanone (1), gallic acid (2), 3,4,8,9,10-pentahydroxydibenzo [b, d] pyran-6-one (3), epicatechin (4), catechin (5), kaempferol-3-O-robinobioside (6), apigenin (7), 2-hydroxysuccinic acid (8), 5-hydroxymethylfuroic acid (9), gallic acid methyl ester (10), caffeic acid (11), tanshinol (12), diolmycin B2 (13), oresbiusin A (14), 3,4-dihydroxyphenyl-β- D-glucopyranoside (15), and (2S)-pyroglutamic acid (16). Conclusion Compounds 7-16 are obtained from the genus of Mallotus for the first time, and compounds 4-16 are obtained from M. furetianus for the first time.
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Objective To establish the quality standard for YI SHI MINGMU granules.Methods TLC method was used to identify Lycium barbarum L and Salvia miltiorrhiza.HPLC method was used to quantitatively analyze the concentration of Tanshinol sodium.The analysis was carried out on a column of Kromasil C18(4.6 mm×150 mm,5 μm)with a mobile phase of methanol and 0.5 % acetic acid at the flow rate of 0.4 ml/min.Column temperature was 30 ℃.The detection wavelength was 280 nm.Results The linearity range of Tanshinol sodium was 2.00~60.00 μg/ml ,r2=0.999 7(n=6),with the average recoveries of 105.62%, RSD=1.60%.Conclusion This method is accurate, simple, reliable and reproducible.It can be used for the quality control of YI SHI MINGMU granules.
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Objective To develop a method for simultaneous determination of three hydrophilic components and two lipophilic components in Radix et Rhizoma Salviae Miltiorrhizae. Methods The RP-HPLC method was performed by using a Welchrom C18 column(250 mm×4.6 mm,5 μm)with a mobile phase of acetonitrile (A)-0.1%phosphoric acid(B). The gradient elution program was as follows:0-15 min,10%→12%A;-35 min,12%→20%A;-45 min,20%→60%A;-65 min,60%→65%A;-80 min,65%→80%A;-90 min,10%A. The flow rate was kept at 1.0 mL?min-1 . The detection wavelength was set at 280 nm. The column temperature was 30 ℃ . Results A good linearity was obtained over 0.059 5-2.380 0 μg for tanshinol, 0.346 0-13. 840 0 μg for rosmarinic acid, 0. 656 0 - 26. 240 0 μg for salviamolic acid B, 0. 420 0 - 16.800 0 μg for cryptotanshinone and 0.414 0- 16.560 0 μg for tanshinoneIIA, respectively ( r = 0.999 9). The average recovery rates were between 98.69%-100.91% with RSD less than 1.2%(n = 6). Conclusion The method is rapid, accurate, credible and repeatable, and can provide basis for the quality control of Radix et Rhizoma Salviae Miltiorrhiza.
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Salvia miltiorrhiza is a traditional Chinese medicine for the treatment of cardiovascular diseases .Recently, increasing evidence demonstrates that the water-soluble compounds isolated from Salvia miltiorrhiza,including tanshinol and salvianolic acid B, exert a regulatory influence on bone metabolism .The under-lying mechanism of these compounds involves various pathways , such as Wnt/β-catenin, ERK, BMP, OPG/RANKL/RANK and FoxO mediated oxidative stress pathway .This paper reviews pre-vious effects and mechanism of polyphenolic acids in Salvia milt-iorrhiza , which may provide the base for the research and devel-opment of the new agents to treat osteoporosis .
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Objective To explore the effect of heating temperature and heating time on the dissolution characteristics of phenolic acids in water extract of Danshen Decoction. Methods The contents of tanshinol, protocatechuic aldehyde and salvianolic acid B in water extract of Danshen Decoction were determined by reverse-phase high performance liquid chromatography (RP-HPLC) . The changes of the contents were monitored by classic constant temperature acceleration method. Results Of the three components, the content of salvianolic acid was decreased with the increase of heating temperature and the prolongation of heating time, and the degradation characteristics of the salvianolic acid B met the first-order kinetics equation. Conclusion The water-soluble components have less stability in the water extracts of Danshen Decoction. Lignum Santali Albi and Fructus Amomi accelerate the degradation speed of Radix Salviae Miltiorrhizae, suggesting that we should pay attention to the influence of heating temperature and heating time on the content of salvianolic acid B in the extraction and concentration process.
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Aim To investigate the effect of tanshinol on bone mineral density and microstructure of proximal tibias in rats with bone loss induced by glucocorticoid. Methods Sixty 7-month-old female SPF SD rats were randomly divided into 6 groups with 1 0 rats per group:control group(saline:5 ml·kg -1 ·d -1 ),glucocorti-coid group (prednisone acetate:6 mg·kg -1 ·d -1 ), glucocorticoid +low dose of tanshinol group(1 2.5 mg ·kg -1 ·d -1 ),glucocorticoid +medium dose of tan-shinol group (25 mg·kg -1 ·d -1 ),glucocorticoid +high dose of tanshinol group (50 mg·kg -1 ·d -1 ), glucocorticoid +(positive control drug)calcitriol group (0.045 μg · kg -1 · d -1 ).Rats were gavaged with prednisone acetate continuously for 1 4 weeks to estab-lish the bone loss model.Meanwhile,tanshinol and calcitriol were orally administered to the rats which were treated with prednisone acetate for intervention. At the end of the experiment,the left proximal tibias were collected for Micro-CT scanning and three-dimen-sional reconstruction of cortical and trabecular bone re- spectively to observe the changes of bone microstruc-ture and test related parameters.Results Bone min-eral density was decreased and bone microstructure was destroyed in proximal tibias of rats after treatment with glucocorticoid.Both tanshinol (25 mg·kg -1 ·d -1 ) and calcitriol(0.045 μg·kg -1 ·d -1 )could increase bone mineral density and improve bone microstructure in proximal tibias without significant differences be-tween each other.Tanshinol (50 mg · kg -1 · d -1 ) could improve bone microstructure to some extent,but it had no significant effect on bone mineral density. Tanshinol(1 2.5 mg·kg -1 ·d -1 )had no significant effect on bone mineral density or microstructure.Con-clusion Oral administration of tanshinol (25 mg · kg -1 ·d -1 )to the rats treated with glucocorticoid can increase bone mineral density and improve bone micro-structure in proximal tibias.
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Diseases with excessive angiogenesis such as tumors need to be treated with anti-angiogenesis agents, while ischemic diseases need to be treated with pro-angiogenesis agents. Salviae Miltiorrhizae Radix et Rhizoma, the representative drug which promotes circulation and resolves clots, was widely used in treating tumors and ischemic diseases. This review focused on the current research on the angiogenic effect of Salviae Miltiorrhizae Radix et Rhizoma and its water-soluble or fat-soluble components. It was found that there are some controversy reports. Both pro-angiogenic and anti-angiogenic effects of Salviae Miltiorrhizae Radix et Rhizoma and its components have been reported. The major difference between tumor and ischemia is the maturity and stability of vessels. Angiogenesis is a complex process involving many signaling molecules. Therefore, the components of Salviae Miltiorrhizae Radix et Rhizoma will regulate the levels and functions of angiogenesis, and then vessels produce different maturity and stability. Overall, such controversy may be caused by differences in experimental conditions, the diversity of Salviae Miltiorrhizae Radix et Rhizoma components, the complexity of the angiogenic process, and the differences of drug distribution under different pathological states in vivo.
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OBJECTIVE:To establish a method for simultaneous determination of tanshinol,protocatechuic aldehyde,paeoni-florin,ferulic acid and salvianolic acid B in Shenshao oral liquid. METHODS:RP-HPLC was performed on the column of Eclipse XDB C18 with mobile phase of 0.5%phosphoric acid-methanol-acetonitrile(gradient elution)at the flow rate of 1.0 ml/min,the de-tection wavelength was 280,230 and 320 nm,column temperature was 30 ℃ and volume was 10 μl. RESULTS:Under the chro-matographic conditions,5 kinds of components could be completely separated,the linear range of tanshinol,protocatechuic alde-hyde,paeoniflorin,ferulic acid and salvianolic acid B were respectively 24-384 μg/ml(r=0.999 9),1.25-20 μg/ml(r=0.999 9), 40.5-648 μg/ml(r=0.999 8),1.5-24 μg/ml(r=0.999 9),145-2 320 μg/ml(r=0.999 9);RSDs of precision,stability and reproduc-ibility tests were no more than 2.2%;the average recovery was respectively 100.7%(RSD=1.23%,n=9),100.0%(RSD=2.19%,n=9),99.6%(RSD=0.87%,n=9),100.3%(RSD=1.11%,n=9) and 99.3%(RSD=2.46%,n=9). CONCLUSIONS:The method is specific with good precision and reproducibility,and can be used for the content determination of 5 components in Shenshao oral liquid.
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OBJECTIVE:To establish a method for the content determination of tanshinol,protocatechuic aldehyde,salvianolic acid B,paeoniflorin and ferulic acid in Xinning tablets. METHODS:Multi-target with multi-wavelength HPLC was performed on the column of Agilent Zorbax Eclipse XDB-C18 with the mobile phase of acetonitrile-methanol-0.5% H3PO4 aqueous solution(gradi-ent elution) at the flow rate of 1.0 ml/min. The column temperature was 30 ℃,volume was 10 μl and the detection wavelength was 280 nm for tanshinol,protocatechuic aldehyde and salvianolic acid B,230 nm for paeoniflorin and 320 nm for ferulic acid. RESULTS:The linear range was 66.25-1060.00 μg/ml for tanshinol(r=0.999 9),5.55-88.86 μg/ml for protocatechuic aldehyde (r=0.999 9),187.20-2 995.20 μg/ml for salvianolic acid B(r=0.999 7),23.71-379.39 μg/ml for paeoniflorin(r=0.999 9)and 0.20-3.12μg/ml for ferulic acid(r=0.999 7);the RSDs of precision,stability and repeatability tests were all less than 2%;the av-erage recoveries were respectively 98.85%(RSD=0.12%,n=6),97.95%(RSD=0.19%,n=6),99.18%(RSD=0.37%,n=6), 98.14%(RSD=0.25%,n=6) and 97.16%(RSD=1.36%,n=6). CONCLUSIONS:The method is simple and convenient with good separation effect,and can be used for the content determination of the 5 components in Xinning tablets.
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Salvia miltiorrhiza is dry root or rhizome of the labia-tae plant Salvia miltiorrhiza, whose water-soluble ingredients are recognized as its bioactive components in cardiovascular disea-ses. Based on researches at home and abroad in recent years, this article summarizes pharmacological effect resisting myocardi-al ischemia of Salvia miltiorrhiza ’ s water soluble compounds in multi-level from the prevention, treatment and anti-reperfusion damage, to provide theoretical basis for the research and devel-opment of the active monomers in Salvia miltiorrhiza and com-pound preparations.
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OBJECTIVE:To establish a method for the contents determination of tanshinol,protocatechuic aldehyde,ferulic ac-id and salvianolic acid B in Yixinshu capsule. METHODS:Dual-wavelength HPLC was performed on the column of Eclipse XDB-C18 with mobile phase of 0.5%phosphoric acid-methanol-acetonitrile(gradient elution)at the flow rate of 1.0 ml/min,the de-tection wavelength was 280 nm(tanshinol,protocatechuic aldehyde,salvianolic acid B)and 320 nm(ferulic acid),column tempera-ture was 30℃and volume was 10 μl. RESULTS:The linear range of tanshinol,protocatechuic aldehyde,ferulic acid and salvianolic acid B were respectively 9-144μg/ml(r=0.999 9),0.5-8μg/ml(r=0.999 9),0.65-10.4μg/ml(r=0.999 9)and 221.25-3 540μg/ml (r=0.999 9);RSDs of precision,stability and reproducibility tests were no more than 1.90%;the average recovery was respective-ly 100.8%(RSD=1.65%,n=9),100.1%(RSD=2.87%,n=9),100.1%(RSD=3.01%,n=9) and 99.4%(RSD=2.05%,n=9). CONCLUSIONS:The method is simple and reproducible,and can be used for the quality control of Yixinshu capsule.
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Objective To develop an RP-HPLC method for determination of tanshinol, protocatechualdehyde, paeoniflorin, puerarin, ferulic acid, tanshinone II A and astragaloside in blood-invigorating and stasis-removing prescription (BSP). Methods The analysiswas performed with a column of Waters Symmetry Shield™ RP C18(150 mm × 4. 6 mm, 3. 5 μm), and the mobile phase consisted of methanol-0. 25% acetic acid. The flow rate was 0. 8 mL/min and the column temperature was 30°C. UV was employed to determine the contents of tanshinol, protocatechualdehyde, paeoniflorin, puerarin, ferulic acid and tanshinone II A, and the detection wavelength was set at 280 nm. Evaporative light scattering detection (ELSD) was employed to determine the contents of astragaloside. The temperature of drift tube was 90°C and the gas flow was 2. 8 L/min (compressed air). Results The linearity was obtained over 0. 01-0. 80 μg (r = 0. 999 8) for tanshinol, 0. 005-0. 4 μg (r=0. 999 7) for protocatechualdehyde, 0. 05-4 μg (r = 0. 999 8) for paeoniflorin, 0. 005-0. 4 μg (r = 0. 999 7) for puerarin, 0.006-0.5 μg (r=1) for ferulic acid, 0. 005-0. 4 μg (r=1) for tanshinone II A, and 0. 031-2. 46 μg (r= 0. 999 3) for astragaloside. The recoverieswere all between 97. 0%-101. 0%, and RSDs were all less than 2%. The contents (mean) of tanshinol, protocatechualdehyde, paeoniflorin, puerarin, ferulic acid, tanshinone H A and astragaloside in three batches of samples were 1. 15, 0. 13, 4. 48, 0. 80, 0. 72, 0. 31 and 3. 12 mg/g, respectively. Conclusion The method in our study is convenient, accurate and sensitive, and it provides a reference for the determination of active ingredients in BSP.
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OBJECTIVE: To establish an RP-HPLC method for the determination of tanshinol in human plasma.METHODS: The determination was performed on Kromasil-C18(150 mm?4.6 mm,5 ?m).The mobile phase consisted of acetonitrile-potassium dihydrogen phosphate(95∶5,adjusted pH to 2.8 with phosphoric acid) at a flow rate of 1.0 mL?min-1.The detection wavelength was set at 281 nm;the column temperature was set at 25 ℃;and the internal standard was p-aminobenzoic acid.RESULTS: The linear range of tanshinol was 2.72～1 561.58 ng? mL-1(r=0.999 4) and the lowest detection limit was 1.0 ng?mL-1(S/N=4);both the intra-day RSD and inter-day RSD were less than 6%.The methodological recovery was 95.6%.CONCLUSION: The method is sensitive,convenient and accurate,and it is suitable for the determination and pharmacokinetic study of tanshinol in human plasma.
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OBJECTIVE:To consummate the quality control criteria of Lemai granule.METHODS:Qualitation identifica?tion of Radix paeoniae rubra and nutgrass galingale rhizome in Lemai granule were performed by TLC,and the content of tanshinol-the chief component in Radix Salviae miltiorrhizae was determined by HPLC.RESULTS:The results showed that the color spectrum identification of Radix paeoniae rubra and nutgrass galingale rhizome in Lemai granule were positive.Good linear relationship was achieved between tanshinol sodium and peak area when the concentration range of which was0.203?g～2.030?g(r=0.9998).The average recovery rate was98.93%(RSD=1.04%).CONCLUSION:The established method in this research can be used as the quality control for Lemai granule.
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Objective To investigate the effects of tanshinol on the proliferation,apoptosis and NF-?B activation in rat hepatic stellate cells(HSCs) after IL-1? inducement,and to elucidate the anti-fibrotic molecular mechanisms of tanshinol.Methods The rat HSCs was isolated with collagenase in situ liver recirculation perfusion and cultured in vitro.The cells were divided into 5 groups: normal control,IL-1? treatment group(10 ng/ml),and tanshinol group 1,2 and 3.The later 3 groups were pretreated with tanshinol at the concentrations of 0.062 5,0.125 and 0.25 mmol/L respectively followed by 10 ng/ml IL-1? treatment 1 h later.MTT colorimetric assay was used to detect the proliferation of HSCs.AO/EB immunoflurorescence microscopy and combination Annexin-V-FITC/PI double-labelimmunofluorescence with flow cytometer were employed to examine the apoptosis of HSCs.Synthesis and secretion of collagen Ⅲ were detected by the quantitative immunocytochemical assay and ELISA respectively.The amounts of cytoplasm p-I?B? and NF-?B p65,and nuclear NF-?B p65 in HSCs were determined by Western blotting.Immunocytochemical staining and Western blotting were used to observe nuclear translocation of NF-?B p65.Results IL-1? increased the proliferation of HSCs(P
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Objective To study the pharmacokinetics of Danshensu (tanshinol) in Danqi Injection(DQI) in dogs. Methods HPLC-UV detection was used to determine the Danshensu level in biological samples. The Lichrospher C18 column(4.6?250 mm,5?m) was used as an analytical column with a mobile phase consisted of acetonitrile-1 %acetic acid (8∶92),the flow rate being 1.0mL?min-1 and the wave-length being 280 nm. Results After intravenous injection of DQI in dogs,the plasma concentration-time curve of DQI in dogs fitted well to a two-compartment model,with the characteristics of fast absorption and slow elimination.Its pharmacokinetics parameters are as follows:?=1.533 h-1,?=0.5111 h-1,T(peak)=0.25 h,T1/2?=0.573 h. Conclusion The pharmacokinetics of DQI in dogs fits the two-compartment model.In the pharmacokinetics of Danshensu,elimination course is the main course with the slow and lasting characteristics.