Isolation and Identification of Chemical Constituents from Rhizomes of Dendropanax proteus / 中国实验方剂学杂志

The chemical constituents from 95% ethanol extract of Dendropanax proteus rhizomes and their anti-inflammatory activities were investigated. These compounds in 95% ethanol extract of D. proteus rhizomes were isolated and purified by silica gel column chromatography, medium-pressure liquid chromatography, preparative liquid chromatography, etc. Their structures were elucidated based on the spectral data and physicochemical properties. All the compounds were tested for their ability to inhibit lipopolysaccharide (LPS) induced nitric oxide production in the murine microglia BV2 cell line. Nine compounds were isolated from the ethyl acetate fraction of 95% ethanol extract of D. proteus rhizomes, and identified as (-)-syringaresinol (1), (+)-(7S,8S)-1',4-dihydroxy-3,3',5'-trimethoxy-7',8,9'-trinor-8,4'-oxyneoligna-7,9-diol (2), erythro-guaiacylglycerol-β-O-4'-coniferyl ether (3), threo-guaiacylglycerol-β-O-4'-coniferyl ether (4), coniferyl alcohol (5), 7-O-ethylguaiacylglycerol (6), vanillin (7), syringaldehyde (8), and excoecanol B (9). Compounds 2 and 4 showed neuritis inhibitory activity against microglial inflammation factor, their half inhibitory concentrations (IC50) were 5.85, 7.29 μmol ·L-1, respectively. Compounds 1-6,8-9 are isolated from this plant for the first time, compounds 2 and 4 exhibit the potent inhibitory activity.

Mechanisms of Astragalus Polysaccharids Synergized with Doxorubicin against Drug Resistance in HL-60/A Cells / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To explore the synergistic anti-tumor mechanism of astragalus polysaccharids(APS) combined with doxorubicin in HL-60/A cells.</p><p><b>METHODS</b>The phenotype of HL-60/A was identified by using RT-PCR, Wester blot and CCK-8, the antitumor effect of astragalus polysaccharids combined with doxorubicin on HL-60/A cells was detected by CCK-8, the apoptosis of HL-60/A after treating with different drugs was detected by Annexin-V, the caspase casade activation was detected by Wester blot, the effect of astragalus polysaccharids on expression and function of multidrug resistance protein (MRP) was detected by using real-time quantitative PCR and Western blot.</p><p><b>RESULTS</b>HL-60/A cells displayed obvious characteristics of resistance to doxorubicin, whose resistance were about 27 times that of its sensitive cell line HL-60. In addition, it was further found that astragalus polysaccharids could obviously increase the cell growth inhibition, induce cell apoptosis and caspase cascade activation, decrease the expression of MRP and increase the drug concentration in HL-60/A when combined with doxorubicin.</p><p><b>CONCLUSION</b>Astragalus polysaccharids combined with doxorubicin can effectively overcome the drug resistance of HL-60/A. The mechanism may be associated with decreasing the expression of MRP, inhibiting drug efflux and increasing the intracellular drug concentration, then inducing HL-60/A cell apoptosis.</p>

OATP1B1 in drug-drug interactions between traditional Chinese medicine Danshensu and rosuvastatin / 药学学报

The study was designed to explore the drug-drug interactions mechanisms mediated by OATP1B1 between traditional Chinese medicine Danshensu and rosuvastatin. First, the changes of rosuvastatin pharma­cokinetics were investigated in presence of Danshensu in rats. Then, the primary rat hepatocytes model was established to explore the effects of Danshensu on the uptake of rosuvastatin by hepatocytes. Finally, HEK293T cells with overexpression of OATP1B1*1a and OATP1B1*5 were established using a lentiviral delivery system to explore the effects of Danshensu on the uptake of rosuvastatin. Rosuvastatin pharmacokinetic parameters of Cmax, AUC0-t, AUC0-∞ were increased about 123%, 194% and 195%, by Danshensu in rats, while the CLz/F value was decreased by 60%. Uptake of rosuvastatin in the primary rat hepatocytes was decreased by 3.13%, 41.15% and 74.62%, respectively in the presence of 20, 40 and 80 μmol·L-1 Danshensu. The IC50 parameters was (53.04 ± 2.43) μmol·L-1. The inhibitory effect of Danshensu on OATP1B1 mediated transport of rosuvas­tatin was related to the OATP1B1 gene type. In OATP1B1*5-HEK293T mutant cells, transport of rosuvastatin were reduced by (39.11 ± 4.94) % and (63.61 ± 3.94) %, respectively, by Danshensu at 1 and 10 μmol·L-1. While transport of rosuvastatin was reduced by (8.22 ± 2.40) % and (11.56 ± 3.04) % and in OATP1B1*1a cells, respectively. Danshensu significantly altered the pharmacokinetics of rosuvastatin in rats, which was related to competitive inhibition of transport by OATP1B1. Danshensu exhibited a significant activity in the inhibition of rosuvastatin transport by OATP1B1*5-HEK293T, but not by OATP1B1*1a, suggesting a dependence on OATP1B1 sequence.

OATP1B1 in drug-drug interactions between traditional Chinese medicine Danshensu and rosuvastatin / 药学学报

The study was designed to explore the drug-drug interactions mechanisms mediated by OATP1B1 between traditional Chinese medicine Danshensu and rosuvastatin. First, the changes of rosuvastatin pharmacokinetics were investigated in presence of Danshensu in rats. Then, the primary rat hepatocytes model was established to explore the effects of Danshensu on the uptake of rosuvastatin by hepatocytes. Finally, HEK293T cells with overexpression of OATP1B1*a and OATP1B1*5 were established using a lentiviral delivery system to explore the effects of Danshensu on the uptake of rosuvastatin. Rosuvastatin pharmacokinetic parameters of C(max0, AUCO(0-t), AUC(0-∞) were increased about 123%, 194% and 195%, by Danshensu in rats, while the CL z/F value was decreased by 60%. Uptake of rosuvastatin in the primary rat hepatocytes was decreased by 3.13%, 41.15% and 74.62%, respectively in the presence of 20, 40 and 80 μmol x L(-1) Danshensu. The IC50 parameters was (53.04 ± 2.43) μmol x L(-1). The inhibitory effect of Danshensu on OATP1B1 mediated transport of rosuvastatin was related to the OATP1B1 gene type. In OATP1B1*5-HEK293T mutant cells, transport of rosuvastatin were reduced by (39.11 ± 4.94)% and (63.61 ± 3.94)%, respectively, by Danshensu at 1 and 10 μmol x L(-1). While transport of rosuvastatin was reduced by (8.22 ± 2.40)% and (11.56 ± 3.04)% and in OATP1B1*1a cells, respectively. Danshensu significantly altered the pharmacokinetics of rosuvastatin in rats, which was related to competitive inhibition of transport by OATPJBI. Danshensu exhibited a significant activity in the inhibition of rosuvastatin transport by OATP1B1*5-HEK293T, but not by OATP1B1*1a, suggesting a dependence on OATP1B1 sequence.

Effect of Ursolic Acid on Breast Cancer Resistance Protein-mediated Transport of Rosuvastatin In Vivo and Vitro / 中国医学科学杂志(英文版)

<p><b>OBJECTIVE</b>To evaluate whether ursolic acid can inhibit breast cancer resistance protein (BCRP)-mediated transport of rosuvastatin in vivo and in vitro.</p><p><b>METHODS</b>Firstly, we explored the pharmacokinetics of 5-fluorouracil (5-FU, a substrate of BCRP) in rats in the presence or absence of ursolic acid. Secondly, we studied the pharmacokinetics of rosuvastatin in rats in the presence or absence of ursolic acid or Ko143 (inhibitor of BCRP). Finially, the concentration-dependent transport of rosuvastatin and the inhibitory effects of ursolic acid and Ko143 were examined in Madin-Darby Canine Kidney (MDCK) 2-BCRP421CC (wild type) cells and MDCK2-BCRP421AA (mutant type) cells.</p><p><b>RESULTS</b>As a result, significant changes in pharmacokinetics parameters of 5-FU were observed in rats following pretreatment with ursolic acid. Both ursolic acid and Ko143 could significantly affect the pharmacokinetics of rosuvastatin. The rosuvastatin transport in the BCRP overexpressing system was increased in a concentration-dependent manner. However, there was no statistical difference in BCRP-mediated transport of rosuvastatin betweent the wild type cells and mutant cells. The same as Ko143, ursolic acid inhibited BCRP-mediated transport of rosuvastatin in vitro.</p><p><b>CONCLUSION</b>Ursolic acid appears to be a potent modulator of BCRP that affects the pharmacokinetic of rosuvastatin in vivo and inhibits the transport of rosuvastatin in vitro.</p>

The cytotoxicity of indirubin derivative PHII-7 against human breast cancer MCF-7 cells and its mechanisms / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To observe the cytotoxicity of indirubin derivative PHII-7 against human breast cancer MCF-7 cells and to study its primary mechanisms.</p><p><b>METHODS</b>The proliferation of MCF-7 cells was detected using MTT colorimetry. Annexin V/PI double staining was applied to detect the apoptosis rate of MCF-7 cells. The distribution of cell cycles was detected using PI staining and flow cytometry (FCM). The levels of reactive oxygen species (ROS) in MCF-7 cells were detected by DCFH-DA staining. The mRNA and protein levels of c-fos were detected using RT-PCR and Westem blot analysis.</p><p><b>RESULTS</b>PHII-7 at different concentrations inhibited the proliferation of MCF-7 cells in a concentration-dependent manner, with the inhibitory rate ranging from 43.13% to 90.90% (P < 0.05). The inhibition was strengthened along with increased concentrations. PHII-7 at different concentrations could induce the apoptosis of MCF-7 cells. The early apoptosis rate was 1.43% +/- 0.02%, 9.14% +/- 0.36%, and 45.79% +/- 8.46%, respectively with the action of 1.25, 2.50, and 5.00 micromol/L PHII-7, respectively, showing dose-dependent manner. FCM analysis found that the proportion of MCF-7 cells in the G0/G1 phase and the S phase decreased after treatment with PHII-7, and the ratio of MCF-7 cells in the G2/M phase obviously increased (P < 0.01). The intra-cellular ROS level was significantly elevated 2 h after pretreatment with PHII-7. The levels of the protooncogene c-fos mRNA and protein were down-regulated in a dose-dependent manner after action of PHII-7.</p><p><b>CONCLUSIONS</b>PHII-7 exerted obvious in vitro cytotoxic effects on MCF-7 cells. Its mechanisms might be associated with arresting the cell cycle, regulating the redox equilibrium, and down-regulating the expression of the protooncogene.</p>

Effects of rhBNP on left ventricular remodeling in rats with acute myocardial infarction / 中华心血管病杂志

<p><b>OBJECTIVE</b>To assess the effects of rhBNP on left ventricular (LV) remodeling in rats with acute myocardial infarction (AMI).</p><p><b>METHODS</b>AMI was induced by ligating coronary artery in male Sprague Dawley rats. Two days after surgery, AMI rats received intravenous infusion of rhBNP (15 microg/kg or 5 microg/kg once daily, n = 15 each) or saline (placebo control, n = 15) through Jugular Vein. Sham-operated rats (n = 15) served as normal control. Four weeks later, hemodynamic measurements were performed, left ventricular weight (LVW), ratio of left ventricular weight to body weight (LVW/BW), left ventricular diameter (LVD) and infarct size were determined. Plasma angiotensin II and myocardial angiotensin II levels were also measured.</p><p><b>RESULTS</b>Compared with sham-operated rats, LVW, LVW/BW, LVD and myocardial angiotensin II level were significantly increased, while the LV systolic pressure (LVSP), +/- dp/dt were significantly reduced in saline treated AMI rats (all P < 0.05). LVW/BW, MI size, LVD and myocardial angiotensin II in rhBNP treated AMI rats were significantly lower [LVW: (492.6 +/- 34.0) mg, (498.8 +/- 47.8) mg, (570.0 +/- 24.2) mg, P < 0.01; LVW/BW: 2.0 +/- 0.2, 2.0 +/- 0.2, 2.3 +/- 0.1, P < 0.01; LVD: (25.3 +/- 2.9)%, (31.4 +/- 3.0)%, (46.4 +/- 3.0)%, P < 0.01; myocardial angiotensin II: (881.3 +/- 62.7) pg/L, (1186.0 +/- 94.5) pg/L, (2436.7 +/- 280.3) pg/L, P < 0.05], while LVSP and +/- dp/dt in rhBNP treatment groups were significantly increased than saline treated AMI rats (P < 0.05 or P < 0.01).</p><p><b>CONCLUSION</b>RhBNP is effective in attenuating left ventricular remodeling after AMI in rats.</p>