To evaluate whether Shenfu injection (SFI) protects against cardiac myocyte injury induced by Fupian injection (FPI) in vitro.

Objective E0703 is derived from estradiol and has anti-radiation effects on irradiated mice, the effects of E0703 on cell viability,cell cycle and the related mechanism in irradiated AHH-1 cells were investigated in this study.Methods Cell viability,cell apoptosis and cell cycle were determined by Cell Counting Kit-8.Annexin V-FITC double staining kit and Flow Cytometry,respectively.Level of mRNA and protein were detected by RT-PCR and Western blot,respectively.Results 3.0 Gy 60 Co γ-rays induced significant cell cycle arrest,necrosis,apoptosis and reduction of viability.Pretreatment with E0703 for 12 h before irradiation could increase cell viability and regulate cell cycle.but had no obvious effect on cell necrosis and apoptosis.The expression of cell cycle arrest related molecules Rb and MDM2 were increased after 3.0 Gy irradiation,but decreased significantly with pretreatment of E0703.Moreover.the phosphorylation of Rb was also increased.Conclusions The radioprotective function of E0703 might be exerted through influencing on cell cycle and promoting cell proliferation instead of apoptosis or necrosis.

OBJECTIVE Tostudythehepatotoxicityofveratrinehydrochloride(VH)anditsmecha-nismoninductionofapoptosisinvitro.METHODS HepG2cellswereexposedtoVH0.1-0.6g·L-1 for 24 h,cell viability was examined by CCK-8 assay,and the morphologic changes in HepG2 cells were quantified.After the treatment with VH 0.1 -0.5 g·L-1 for 24 h,cell membrane injury was examined by detecting the release rate of lactate dehydrogenase (LDH).The effect on reactive oxygen species (ROS),mitochondrial membrane potential and apoptosis was detected by flow cytometry.The mRNA expression of p53,Bax,cytochrome c,caspase 9,caspase 3 was evaluated by real-time PCR. RESULTS HepG2cellviabilitywassignificantlyreducedfollowingexposuretoVH0.1-0.5g·L-1. The IC50 value was 0.4 g·L-1 .The 95%confidence limit was 0.2558-0.6965 g·L-1 .The LDH release rate,ROS and apoptosis rate of HepG2 cells were significantly increased after exposure to VH 0.1 -0.5 g·L-1 for 24 h (P<0.05,P<0.01 ),and the mitochondrial membrane potential markedly declined (P<0.05,P<0.01 ).The expression of p53,Bax,cytochrome c,caspase 9 and caspase 3 was increased(P<0.05,P<0.01).CONCLUSION VHhascytotoxicpotential.Damagetocell me mbrane and mitochondria and initiation of apoptosis-related genes of caspase 9 and caspase 3 mRNA expression may be the mechanis m of apoptosis.

Aim To investigate the induction effect of ginsenoside Rc, Re, Rf and Rg1 on CYP1A1, and further validate the role of aryl hydrocarbon receptor in CYP1A1 expression. Methods Dual luciferase re-porter gene system was performed. Four kinds of gin-senoside were screened for aryl hydrocarbon receptor activation by reporter assays, and TCDD as the positive control. Further with different concentrations of ginsen-oside Rc, Re, Rf and Rg1 treated on LS174T cells, RNA and total protein were extracted to detect the reg-ulating effect of ginsenosides on CYP1 A1 mRNA and protein expression with Real-time PCR and Western blot technology respectively. Results Reporter gene screening showed that the ginsenoside Rc, Re, Rf and Rg1 could activate AhR and had potential effects on the induction of CYP1A1 enzyme. Meanwhile, dose-de-pendent induction of the gene expression were observed in response to ginsenoside Rc, Re, Rf and Rg1 and the levels of CYP1 A1 protein expression were increased by ginsenoside Rc, Re, Rf and Rg1 in varying de-grees. Conclusion Ginsenoside Rc, Re, Rf and Rg1 can up-regulate the gene and protein expression of CYP1 A1 possibly via the AhR-mediated CYP1 A1 path-way.

Aim To investigate the effects of Fufangdanshen on TNF-? stimulated vascular smooth muscle cells(VSMC) and discover potential molecular mechanism to cure atherosclerosis at molecular level.Methods TNF-? stimulated VSMC was used as dysfunction cell model;VSMC proliferation was detected with MTT and FACS;proteomic protocol involving 2-DE,image analysis and spectrometry detection were used to detect regulated protein by Fufangdanshen.Results Fufangdanshen inhibited expression of 16 proteins but promoted expression of 24 proteins in TNF-? stimulated VSMC.The expression of CaMKK and N-ras were decreased by Fufangdanshen,which farther decreased matrix metalloproteinase production;while cell cycle related protein p21,p53 were increased by Fufangdanshen in TNF-? treated VSMC.Conclusion The inhibition of proliferation and migration of VSMC through decreased matrix metalloproteinase production and increased cell cycle related protein p21,p53 might be the mechanism of Fufangdanshen to cure atherosclerosis.

An ultra-performance liquid chromatography coupled with time-of-flight mass spectrometry (UPLC/Q-TOF-MS)-based chemical analytic technology was used to evaluate the chemical constitution of Radix Aconiti Lateralis Preparata in the process of decocting, so as to provide a scientific basis for processing Radix Aconiti Lateralis Preparata.

In the present study, an ultra performance liquid chromatography coupled with time-of-fight mass spectrometry (UPLC-TOF/MS) based chemical profiling approach was used to evaluate chemical constitution between co-decoction and mixed decoction of ginseng and Radix Aconiti Praeparata. Two different kinds of decoctions, namely co-decoction of ginseng and Radix Aconiti Praeparata: water extract of mixed two herbs, and mixed decoction of ginseng and Radix Aconiti Praeparata: mixed water extract of each individual herbs, were prepared. Batches of these two kinds of decoction samples were subjected to UPLC-TOF/MS analysis. The datasets of t(R) m/z pairs, ion intensities and sample codes were processed with supervised partial least squared discriminant analysis (OPLS-DA) to holistically compare the difference between these two decoction samples. Significant difference between the two decoction samples was showed in the results of positive ion mode. The contents of hypaconitine and deoxyaconitine decreased, while that of benzoylmesaconine, benzoylhypaconine and dehydrated benzoylmesaconine increased in the samples of co-decoction of ginseng and Radix Aconiti Praeparata. The content of diester-diterpenoid alkaloids decreased, while that of monoester-diterpenoid alkaloids increased, which is probably the basis of toxicity-attenuated action when combined ginseng with Radix Aconiti Praeparata.

Aim To observe the cytochrome 450 effect of ginsenoside Re on H9c2 cells, in order to clarify the molecular mechanism of ginsenoside Re. Methods H 9 c 2 cells were separately treated with ginsenoside Re for 1, 5, 10, 50, 100 μmol·L-1 or 6, 24, 36, 48, 60 h. CYP2C11, 2J3, 4A1, 4A3, 4F4 and ANP mR-NA expressions were analyzed by Real time PCR, and CYP4 A1 , 2 J3 protein expressions were detected by Western blot. Results Compared with control group, ginsenoside Re could effectively upregulate CYP2 C11 , CYP2 J3 , ANP mRNA expression to 1. 6 , 1. 8 , 3. 2 fold, and downregulate CYP4A1, CYP4A3, CYP4F4 mRNA expression to 0. 4, 0. 15, 0. 3 fold. Ginsen-oside Re could decrease CYP4 A1 protein expression in a concentration-dependent manner, while ginsenoside Re could increase CYP2 J3 protein expression in a con-centration-dependent manner. Conclusion Ginsen-oside could regulate CYP450 enzyme and change ANP gene expression, which might be the molecular mecha-nism of ginsenoside Re.

Aim To study the induction effect of Ginkgolide B on CYP3A4,and further verify the role of pregnane X receptor in CYP3 A4 induction expres-sion. Methods With different concentrations of Ginkgolide B treatment on LS174T cells,the CYP3A4 mRNA expression was detected by Q-PCR assay,fur-ther PXR-CYP3 A4 stable translation HepG2 cell lines were used to test the effect of Ginkgolide B on activity of PXR by reporter gene screening assay.CYP3A4 protein expression was detected by Western blot.PXR was knocked down with transfected with siRNA, CYP3 A4 mRNA and protein were detected in the con-dition of PXR low expression.Results The results revealed that the level of CYP 3 A 4 gene and protein expression were significantly increased by Ginkgolide B,and there was no induction effect on PXR.Reporter gene screening showed that Ginkgolide B could en-hance the transcriptional activity of PXR in a concen-tration-dependent manner.Under conditions of low ex-pression of PXR ,Ginkgolide B could also increase ex-pression of CYP3A4,but the induction folds were low-er than those of normal PXR group.Conclusion Ginkgolide B can signicantly up-regulate CYP3 A4 ex-pression via the PXR-CYP3 A4 pathway,and it has no effects on PXR gene expression.

Aim To study the effect of ginsenoside F1 on the enzyme activity and expression of gene of CYP3 A4 through activation of pregnane X receptor ( PXR ) . Methods With different concentrations of ginsenoside F1 treated on LS174T cells, the expression of CYP3A4 mRNA was determined by Q-PCR, and the enzyme activity was measured by P450-GloTM CYP3A4 assay according to the manufacturer′s instructions, fur-ther PXR-CYP3 A4 stable translation HepG2 cell lines were used to test ginsenoside F1 activates PXR by re-porter gene screening assay. Results The results re-vealed that the levels of CYP3 A4 gene and protein ex-pression were significantly increased by ginsenoside F1 in a concentration-dependent manner. At the same time, reporter gene screening showed that ginsenoside F1 could also enhance the transcriptional activity of PXR. Conclusion Ginsenoside F1 can significantly up-regulate the gene expression and enzyme activity of CYP3A4 via the PXR-CYP3A4 pathway.