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Aim To investigate the role of mitochondrial translocator protein(TSPO)in the apoptosis of HepG2 cells induced by tanshinone IIA(Tan II A)and the involved mechanism. Methods Following the HepG2 cells treated with Tan ⅡA at 2.5, 5 and 10 μmol·L-1, the cell viability was determined by MTT assay, and intracellular ATP content was determined by luciferin-luciferase method. Oxygen utilization was measured polarographically with a Clark oxygen electrode. Cell apoptosis was determined by Hoechst 33342 staining and flow cytometry. The mitochondrial membrane potential was assessed with JC-1 staining. The intracellular distribution of TSPO was examined by TSPO immunostaining, and the expressions of TSPO, Cyto C, caspase-3, caspase-9 were determined by immunoblotting analysis. Results Tan II A inhibited the proliferation of HepG2 cells in a dose-and time-dependent manner. The treatment with Tan II A inhibited ATP production and oxygen utilization of mitochondria. In addition, Tan ⅡA enhanced TSPO expression and accumulation in nuclei and up-regulated the expression of Cyto C, caspase-3 and caspase-9. Conclusions Tan II A induces the apoptosis of HepG2 cells, which may be related to the TSPO-mediated mitochondrial dysfunction.
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Aim To explore the effect of tanshinone II A (Tan II A) on reverse cholesterol transport in atherosclerosis model mice and RAW264. 7 cells and the underlying mechanism. Methods Thirty-two male LDLR -/- mice were randomly divided into four groups. These mice were fed with normal diet or high fat diet for 12 weeks. The control group and model group were given normal saline. Tan II A group and atorvastatin group were given Tan II A solution and atorvastatin solution for 12 weeks. RAW264. 7 cells were induced with oxidized low-density lipoprotein (ox-LDL) 100 mg • L-
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Aim To establishan inflammatory model of mouse monocyte macrophages ( RAW264. 7) using li-popolysaccharide (LPS), and to investigate the antiinflammatory activity and mechanism of tanshinone II-A (Tan IIA). Methods Cell viability was determined by CCK-8 method. Cell migration was detected by Tr-answell apparatus. TNF-α, IL-6, IL-1 p, MCP-1 content in cell supernatant was analyzed using ELISA method. The protein expression of MMP-2, MMP-9, TLR4, κB-α, p-κB-α, NFκB and p-NFκB in RAW264.7 cells was investigated by Western blot. Results Tan IIA significantly inhibited .the secretion of TNF-α, IL-6, IL-1β and MCP-1 in LPS induced RAW264.7 cell culture medium , and significantly down-regulated the expression of matrix metalloprotein-ase-2 (MMP-2), MMP-9, TLR4, p-κB-α and p-NFκB , inhibited IκB-α phosphorylation and NFκB entry into the nucleus and activation. Conclusion Tan IIA can inhibit the release of inflammatory factors through the regulation of TLR4/IκB-α/NFκB signaling pathway.
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Liver diseases is one of serious public health problems due to their high global prevalence and poor long-term clinical outcomes. As a main active ingredient extracted from the traditional Chinese medicine Salvia miltiorrhiza, Tanshinone II A (Tan IIA) has been widely studied because of its various biological activities. Many experimental and clinical studies have demonstrated that Tan IIA can prevent and slow down the progression of various diseases through anti-inflammation, anti-oxidation, anti-cancer, anti-angiogenesis and other ways. Moreover, it can treat a variety of diseases including cardiovascular and cerebrovascular diseases, liver diseases, cancerand neurodegenerative diseases. However, Tan IIA is a highly hydrophobic compound with the disadvantages of poor oral absorption and bioavailability. To improve the current applications of Tan IIA, it is necessary to prepare new drug delivery systems such as nanoparticles, polymeric micelles, solid dispersions. In this article, an overview of Tan H A will be given with emphasis on the therapeutic activity and newly prepared dosage forms in liver diseases, including inflammation-related liver damage, liver fibrosis, nonalcoholic fatty liver disease(NAFLD)and hepatocellular carcinoma (HCC). This review may provide theoretical clues for clinical therapeutic applications of Tan IIA in liver diseases.
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Objective: To observe the inhibitory effect of tanshinone II A (Tan II A) on the proliferation of liver cancer HepG2 cells and its inductive effect on the migration and apoptosis of HepG2 cells, and to explore the possible mechanism Methods: The human liver cancer HepG2 cells were cultured in vitro. The HepG2 cells were divided into blank group and different concentrations of Tan II A groups. The cells in different concentrations of Tan II A groups were added with Tan II A at the final concentrations of 0. 5, 1. 0, 2. 0, 5.0 and 10. 0 mg middot; L-1 Tan II A and cultured for 24 h. The morphology of HepG2 cells in various groups was observed under inverted microscope. The inhibitory rates of proliferation of HepG2 cells in various groups were detected by MTT assay. The migration of HepG2 cells in various groups were evaluated by cell scratch assay. The expression levels of nuclear factor-κB (NF-κB) and matrix metalloproteinase-9 (MMP-9) mRNA were detected by RT-PCR method. Flow cytometry was used to detect the percentages of HepG2 cells at different cell cycles in various groups, and the apoptotic rates of HepG2 cells in various groups were detected by TUNEL method. Results: The morphology of HepG2 cells in different concentrations of Tan II A groups were changed. Compared with blank group, the cells in 1. 0 and 2. 0 Tan II A groups showed shrinkage, scattered connection and poor growth, and the inhibitory rates of proliferation of HepG2 cells in 1. 0, 2. 0, 5.0, and 10. 0 mg middot; L-1 Tan II A groups were significantly increased (P< 0. 05) in a dose-dependent manner. The cell scratch assay results showed that with the increasing of Tan E A concentration, the migration number of the cells in 2. 0 and 5. 0 mg middot; L-1 Tan II A groups were decreased significantly. Compared with 0. 5 mg middot; L-1Tan II A group, the expression levels of MMP-9 and NF-κB mRNA in the HepG2 cells in 1. 0, 2. 0, and 5. 0 mg middot; L-1Tan II A groups were decresed (P<0. 05 or P<0. 01). The flow cytometry results showed that compared with blank group, the percentages of HepG2 cells in S phase in 1. 0, 2. 0, 5. 0, and 10. 0 mg middot; L-1Tan II A groups were decreased (P<0. 05), and the percentages of HepG2 cells in G0/G1 and G2 phases were increased (P<0. 05); the TUNEL results showed that compared with blank group, the apoptotic rates of HepG2 cells in 0. 5, 1. 0, 2. 0, 5. 0, and 10. 0 mg middot; L-1Tan II A groups werer increased (P< 0. 05 or P<0. 01). Conclusion: Different concentrations of Tan II A could significantly inhibit the proliferation and migration of human liver cancer HepG2 cells, and induce the apoptosis in a dose-dependent manner; its mechnasim may be related to the inhibition of the expressions of NF-κB and MMP-9 mRNA.
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Objective: To explore the influence of tanshinone 11 A on the expressions of molecular chape rone Cosmc. advanced oxidation protein products (AOPP) in the kidney tissue of the allergic purpura nephritis mice and the levels of extracellular signal-regulated kinase/mitogen activated protein kinase (ERK/MAPK) signaling pathway proteins, and to clarify mechanism of tanshinone 11 A in the treatment of allergic purpura nephritis. Methods: A total of 38 mice were divided into control group∗ model group (allergic purpura nephritis model) and tanshinone 11 A group (allergic purpura nephritis model • tanshinone 11 A treatment), with 12 in each group; the other 2 mice were used to verify the success of modeling. The urine red blood cell counts and urine protein levels of the mice in various groups were measured, and the pathomorphology of kidney tissue of the mice in various groups were observed by periodate schiff-reaction (PAS) staining. The AOPP protein levels in the kidney tissue of the mice were determined by enzyme linked immunosorbent assay (ELISA), and the ERK and phosphorylated ERK (p-ERk) protein expression levels were determined by Western blotting method. The expression levels of ERK. p-ERK and Cosmc mRNA in kidney tissue of the mice were determined by RT-PCR. Results: In control group, there was no hyperplasia of glomerular basement membrane and no glomerular sclerosis; in model group, the proliferation of glomerular basement membrane was obvious∗ the inflammatory cells were infiltrated, and glomerular sclerosis appeared; in tanshinone 11 A group, glomerular basement membrane hyperplasia and inflammatory infiltration were not obvious in the mice, there was only small amount of infiltrated inflammatory cells. Compared with control group, the urine red blood cell count, the 24 h urine protein level, the p ERK mRNA and AOPP protein levels of the mice in model group and tanshinone 11 A group were increased ( P<0. 05 or P<0. 01); the Cosmc mRNA expression levels were decreased (P<0. 01). Compared with model group, the urine red blood cell count. 24 h urine protein level, the p-ERK and AOPP protein levels in the kidney tissue of the mice in tanshinone 11 A group were decreased (P<0. 05 or P<0. 01). and the Cosmc mRNA expression level was increased (P
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Aim To elucidate the effect of rifampin and tanshinone II A on BSEP transport capacity using pravastatin as the BSEP substrate in sandwich-cultured rat hepatocytes (SCRH). Methods SCRH model was established. The doses of drugs were determined by MIT. A HPLC-MS/MS method was developed and was conducted method validation to detect the concentration of pravastatin. The effect of rifampin and tanshinone D A on the concentration of pravastatin in the bile duct was investigated. And the biliary excretion index ( BEI) was calculated. Results The SCRH model was successfully developed. The appropriate doses of rifampin, tanshinone DA, glibenclamide and pravastatin were determined. A stable and reliable HPLC-MS/MS method for the determination of pravastatin was established Compared with blank control group, rifampin reduced the concentration of pravastatin in the bile duct and the BEI of pravastatin. The high concentration of rifampin caused the steepest downward trends ( P < 0 . 0 1 ) . Compared with high concentration group of rifampin, the concentration of pravastatin in the bile duct and the BEI of pravastatin gradually increased after the combination of rifampin and tanshinone II A, and the effect of high concentration of tanshinone II A was the most significant ( P < 0. 0 1 ) . Conclusions Rifampin could inhibit the function of BSEP in SCRH. The combination of tanshinone D A and rifampin could reverse the inhibitor)' effect of BSEP transport capacity caused by rifampin.
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Cardiac dysfunction, a common consequence of sepsis, is the major contribution to morbidity and mortality in patients. Sodium tanshinone IIA sulfonate (STS) is a water-soluble derivative of Tanshinone IIA (TA), a main active component of Salvia miltiorrhiza Bunge, which has been widely used in China for the treatment of cardiovascular and cerebral system diseases. In the present study, the effect of STS on sepsis-induced cardiac dysfunction was investigated and its effect on survival rate of rats with sepsis was also evaluated. STS treatment could significantly decrease the serum levels of C-reactive protein (CRP), procalcitonin (PCT), cardiac troponin I (cTn-I), cardiac troponin T (cTn-T), and brain natriuretic peptide (BNP) in cecal ligation and puncture (CLP)-induced) septic rats and improve left ventricular function, particularly at 48 and 72 h after CLP. As the pathogenesis of septic myocardial dysfunction is attributable to dysregulated systemic inflammatory responses, several key cytokines, including tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), interleukin-6 (IL-6), interleukin-10 (IL-10) and high mobility group protein B1 (HMGB1), were detected to reveal the possible mechanism of attenuation of septic myocardial dysfunction after being treated by STS. Our study showed that STS, especially at a high dose (15 mg·kg), could efficiently suppress inflammatory responses in myocardium and reduce myocardial necrosis through markedly reducing production of myocardial TNF-α, IL-6 and HMGB1. STS significantly improved the 18-day survival rate of rats with sepsis from 0% to 30% (P < 0.05). Therefore, STS could suppress inflammatory responses and improve left ventricular function in rats with sepsis, suggesting that it may be developed for the treatment of sepsis.
Subject(s)
Animals , Female , Humans , Male , Rats , C-Reactive Protein , Genetics , Allergy and Immunology , Cecum , General Surgery , Drugs, Chinese Herbal , Chemistry , Heart , Interleukin-6 , Genetics , Allergy and Immunology , Ligation , Myocardium , Allergy and Immunology , Phenanthrenes , Chemistry , Punctures , Salvia miltiorrhiza , Chemistry , Sepsis , Drug Therapy , Allergy and Immunology , Troponin T , Genetics , Allergy and Immunology , Tumor Necrosis Factor-alpha , Genetics , Allergy and ImmunologyABSTRACT
<p><b>OBJECTIVE</b>To systematically evaluate the effectiveness and safety of Sodium Tanshinone II A Sulfonate Injection (STS) as one adjuvant therapy for treating unstable angina pectoris (UAP).</p><p><b>METHODS</b>Randomized controlled trials (RCTs) of UAP treated by STS were searched in the China National Knowledge Infrastructure Database (CNKI), VIP Database for Chinese Technical Periodicals (VIP), Wanfang Database, the Chinese Biomedical Literature Database (CBM), Web of Science, the Cochrane Library, Embase, and PubMed, which from inception to January, 2016. The Cochrane Risk Assessment Tool was used to evaluate the methodological quality of the RCTs. The Review Manager 5.3 software was used to conduct the metaanalysis.</p><p><b>RESULTS</b>The results showed that 17 RCTs involving 1,372 patients were included. The meta-analysis indicated that the combined use of STS and Western medicine (WM) in the treatment of UAP can obviously improve the total effective rate [risk ratio (RR)=1.31, 95% confidence interval (CI) (1.24,1.39), P<0.0001], and the total effective rate of electrocardiogram [RR=1.43, 95% CI (1.30,1.56), P<0.0001], decrease the level of CRP [mean difference (MD)=-3.06, 95%CI (-3.85,-2.27), P<0.00001], fibrinogen [MD=-1.03, 95% CI (-1.16,-0.89), P<0.00001], and whole blood high shear viscosity [MD=-0.70, 95% CI (-0.92,-0.49), P<0.00001]. Additionally, the occurrence of adverse drug reaction of the experimental group was significantly higher than that of the control group [RR=3.57, 95% CI (1.28, 9.94), P<0.05].</p><p><b>CONCLUSIONS</b>Compared with WM, the combined use of STS was more effective.</p>
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Cardiac dysfunction, a common consequence of sepsis, is the major contribution to morbidity and mortality in patients. Sodium tanshinone IIA sulfonate (STS) is a water-soluble derivative of Tanshinone IIA (TA), a main active component of Salvia miltiorrhiza Bunge, which has been widely used in China for the treatment of cardiovascular and cerebral system diseases. In the present study, the effect of STS on sepsis-induced cardiac dysfunction was investigated and its effect on survival rate of rats with sepsis was also evaluated. STS treatment could significantly decrease the serum levels of C-reactive protein (CRP), procalcitonin (PCT), cardiac troponin I (cTn-I), cardiac troponin T (cTn-T), and brain natriuretic peptide (BNP) in cecal ligation and puncture (CLP)-induced) septic rats and improve left ventricular function, particularly at 48 and 72 h after CLP. As the pathogenesis of septic myocardial dysfunction is attributable to dysregulated systemic inflammatory responses, several key cytokines, including tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), interleukin-6 (IL-6), interleukin-10 (IL-10) and high mobility group protein B1 (HMGB1), were detected to reveal the possible mechanism of attenuation of septic myocardial dysfunction after being treated by STS. Our study showed that STS, especially at a high dose (15 mg·kg), could efficiently suppress inflammatory responses in myocardium and reduce myocardial necrosis through markedly reducing production of myocardial TNF-α, IL-6 and HMGB1. STS significantly improved the 18-day survival rate of rats with sepsis from 0% to 30% (P < 0.05). Therefore, STS could suppress inflammatory responses and improve left ventricular function in rats with sepsis, suggesting that it may be developed for the treatment of sepsis.
Subject(s)
Animals , Female , Humans , Male , Rats , C-Reactive Protein , Genetics , Allergy and Immunology , Cecum , General Surgery , Drugs, Chinese Herbal , Chemistry , Heart , Interleukin-6 , Genetics , Allergy and Immunology , Ligation , Myocardium , Allergy and Immunology , Phenanthrenes , Chemistry , Punctures , Salvia miltiorrhiza , Chemistry , Sepsis , Drug Therapy , Allergy and Immunology , Troponin T , Genetics , Allergy and Immunology , Tumor Necrosis Factor-alpha , Genetics , Allergy and ImmunologyABSTRACT
<p><b>OBJECTIVE</b>To examine whether sodium tanshinone II A sulfonate (STS), the main effective component of Salvia miltiorrhiza is effective in relieving the microcirculatory disturbance of small intestine by suppressing the production of reactive oxygen species (ROS) in rats with sepsis.</p><p><b>METHODS</b>A rat model of sepsis was induced by cecal ligation and puncture (CLP). Rats (n =40) were randomly divided into 4 groups: sham-operated group (sham, n =10), sepsis group (CLP, n =10), STS treatment group (STS, n =10) and ROS scavenger dimethylthiourea (DMTU, n =10) group. Animals in the STS group were injected with STS (1 mg/kg) for 10 min through the right external jugular vein after the CLP operation, and animals in the CLP group were given the same volume of normal saline after the CLP operation. Animals in the DMTU group were intraperitoneally injected with 5 mL/kg of 20% DMTU 1 h before CLP. The histopathologic changes in the intestinal tissues and changes of mesenteric microcirculation were observed. The levels of ROS in intestinal tissues from each group were qualitatively evaluated using a fluorescent microscope. The expressions of apoptosis signal-regulating kinase (ASK1), phosphorylated ASK1 (phospho-ASK1), p38 mitogen-activated protein kinases (p38 MAPK), phosphorylated p38 MAPK (phospho-p38 MAPK) and tissue factor (TF) were determined by Western blotting.</p><p><b>RESULTS</b>It was shown that there were obvious microcirculatory disturbance (P <0.05) and tissue injuries in intestinal tissues after CLP operation. The levels of ROS production, phospho-ASK1, phospho-p38 MAPK and TF were increased. Both STS and DMTU suppressed ROS, phospho-ASK1, phospho-p38 MAPK and TF production, and ameliorated the microcirculatory disturbance and tissues injury (P <0.01).</p><p><b>CONCLUSION</b>STS can ameliorate the microcirculatory disturbance of the small intestine by attenuating the production of ROS in rats with sepsis.</p>
Subject(s)
Animals , Male , Intestine, Small , Pathology , MAP Kinase Kinase Kinase 5 , Metabolism , Microcirculation , Phenanthrenes , Chemistry , Pharmacology , Therapeutic Uses , Phosphorylation , Rats, Wistar , Reactive Oxygen Species , Metabolism , Sepsis , Drug Therapy , Pathology , Thromboplastin , Metabolism , p38 Mitogen-Activated Protein Kinases , MetabolismABSTRACT
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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OBJECTIVE: To observe the effects and mechanisms of sodium tanshinone II A sulfonate (STS) on transforming growth factor-β1 (TGF-β1)-induced atrial fibroblasts differentiation. METHODS: In the culture of neonatal rat atrial fibroblasts, type I/III collagen in the cell culture supernatant were measured by enzyme-linked immunosorbent assay. The proliferation activity was measured by methylthiazolyl tetrazolium(MTT) assay. Atrial fibroblasts differentiation was determined by measuring the expression of α-SMA(α-smooth muscle actin) using Western blot and immunofluorescence. Extracellular signal-regulated kinase 1/2 (ERK1/2) activation was measured by Western blot. RESULTS: STS can decrease TGF-β1-induced elevations of type I/III collagen and proliferation activity; inhibit the expression of α-SMA and ERK1/2 activation. CONCLUSION: SIS can inhibit TGF-β1-induced atrial fibroblasts differentiation, the mechanism may be associated with depression of ERKl/2 signaling pathway.
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OBJECTIVE: To explore the effect of tanshinone II A (Tan II A) on the myocardial apoptosis in rats with sepsis. METHODS: The sepsis rat model was established by cecal ligation and puncture operation (CLP), and the Tan II A was given for 12 consecutive hours. Then, the following indices were measured such as the activities of Na+-K+-ATPase and SOD, the contents of TNF-α, IL-1β, calcium, MDA, apoptotic index and the protein level of Bcl-2, Bax, Fas, caspase-3, calcincurin in myocardial. RESULTS: CD Compared with sham group, CLP treatment can result in decreased myocardial activities of Na+-K+-ATPase and SOD, elevated the contents of IL-1β, calcium, MDA (P<0.05); Tan II A treatment can improve the above aberrant indices. (2) Compared with sham group, CLP treatment can elevated the myocardial apoptotic index (P<0.05), and Tan II A treatment can decrease the elevation of apoptotic index by CLP treatment (P<0.05); (3)Compared with sham group, the protein level of Bax, Fas, caspase-3 and calcincurin significantly increased, and protein level of Bcl-2 decreased (P<0.05) in CLP group, and Tan II A treatment can improve exceptional expression of the above proteins (P<0.05). CONCLUSION: Tanshinone II A shows a protective effect on the myocardial apoptosis in septic rats maybe by depressing inflammatory infiltration and oxidative stress reaction, relieving calcium overload and partly improving the exceptional expression of the proteins such as Bax, Bcl-2, Fas, caspases-3 and calcincurin.
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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 prepare tanshinone II A solid dispersion using spray freeze drying (SFD) method. METHODS: Dissolution profiles of tanshinone II A from different solid dispersions prepared with various excipients were plotted. Based on the dissolution rate, F68 was selected as the excipient. The optimized formula was characterized in terms of morphology, surface area and crystallinity. RESULTS: Compared to raw material, the specific surface of tanshinone II A solid dispersion prepared by SFD method with drug-F68 ratio of 1:9 (w/w) increased by 3 folds. The majority of the tanshinone II A in the solid dispersion transformed from crystalline form to amorphous state. As a result, the dissolution rate of Tanshinone II A significantly enhanced, and 70% of tanshinone II A in the solid dispersion were dissolved in 10 min. CONCLUSION: SFD method has been successfully utilized to prepare solid dispersion of tanshinone II A with enhanced dissolution rate than that of the raw material, which is a prerequisite for increasing its oral bioavailability in vivo. Copyright 2012 by the Chinese Pharmaceutical Association.
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OBJECTIVE: To observe the effects and mechanisms of sodium tanshinone II A sulfonate (STS) on angiotensin II (Ang II)-induced cardiomyocyte oxidative stress. METHODS: In the primary culture of neonatal rat cardiomyocytes, the content of reactive oxygen species (ROS) was measured by 2, 7-dichlorofluorescein diacetate (DCFH-DA). 8-hydroxydeoxyguanosine level in the supernatant was measured by ELISA. As indexes of cardiomyocyte oxidative stress, the cellular contents of MDA and SOD, cell vialibity and LDH release were measured. NADPH oxidase (NOX) activity was measured by chromatometry. The expressions of p47phox was assessed using Western blot. RESULTS: STS can decrease Ang II-induced elevations of ROS level and oxidative stress, and inhibit the expression of p47phox and NOX activity. CONCLUSION: The inhibitory effects of STS on Ang II-induced cardiomyocyte oxidative stress may be associated with depressing NOX signaling pathway via down regulation of p47phox expression. Copyright 2012 by the Chinese Pharmaceutical Association.
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Objective: To select the strains which can produce tanshinone II A 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 II A. 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 II A. The extract from DR12 (outside cell) and DR21 (inside cell) had a peak at retention time identical to that of authentic tanshinone II A in HPLC. Conclusion: The fungi appear to produce the bioactive compound tanshinone II A, and they could be used to produce tanshinone II A by fermentation. It provides a new way to synthesize this natural medicine.
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Objective: To develop a new method for the simultaneous determination of two hydrophilic components and two lipophilic components of Radix et Rhizoma Salviae Miltiorrhizae. Methods: The HPLC-DAD method was employed using a column of Agilent Zorbax TC C18 (4.6 mm × 250 mm, 5 μm) with a mobile phase of methanol -2% acetic acid. The gradient elution program was as follow:0-15 min, 30% B-40% B; 15-20 min, 40% B-60% B; 20-25 min, 60% B-90% B; 25-40 min, 90% B. The detection wavelength was set at 281 nm and the temperature was 35°C. Results: The linearity was obtained over 3.76-120.20 μg · ml-1 (r=0.999 9) for rosmarinic acid, 34.20-109 4.5 μg · ml-1 (r=0.999 9) for salviamolic acid B, 0.64-20.32 μg · ml-1 (r=0.999 9) for clyptotanshinon, and 1.02-32.72 μg · ml-1 (r=0.999 6) for tanshinone II A. The RSDs of precision and stability of the sample were both less than 1% in 48 hours. The average recovery was between 99.72%-100.63%. Conclusion: The present method is simple and has satisfactory efficacy; it can simultaneously determine multiple hydrophilic and lipophilic bioactive components in Salvia miltiorrhiza from different areas.
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Objective: To investigate the influence of tanshinone II A on learning and memory ability, expression of inducible nitric oxide synthase (iNOS), matrix metalloproteinase II (MMP-2) and release of free radicals in the hippocampus of rat Alzheimer's disease (AD) model, and to explore the related mechanism. Methods: Rat AD models were established by direct β-amyloid protein (Aβ) injection method. For tanshinone II A (Tan II A) intervention test, the learning and memory ability of the AD rats were observed, the expressions of iNOS, MMP-2 at gene and protein levels were determined and their correlation were analyzed, and the release of free radical in the hippocampus region of AD rats was quantified. Results: The expressions of iNOS and MMP-2 in the hippocampus region of AD rats were significantly higher than that in the control group (P<0. 05), and the expressions of iNOS and MMP-2 were positively correlated at both mRNA and protein levels (P<0. 05). Meanwhile, nitric oxide (NO), ONOO- and reactive oxygen species (ROS) release in the hippocampus of AD rats were significantly higher than those in the control rats (P<0. 05). The learning and memory ability was obviously decreased in AD model rats. Tan II A intervention (50 mg/kg) significantly reduced the AD-induced release of NO, ONOO- , and ROS, and expression of iNOS (P<0. 05) and MMP-2 protein (P<0. 01) in the hippocampus region; it also greatly improved the learning and memory ability of AD model rats. Conclusion: Tan II A can effectively alleviated the AD symptoms, possibly by inhibiting AD-induced iNOS and MMP-2 expressions, reducing toxic free radical, and suppressing oxidative injury in AD rats.