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1.
Acta Pharmaceutica Sinica ; (12): 2241-2247, 2021.
Article in Chinese | WPRIM | ID: wpr-887050

ABSTRACT

This study investigated the intervention effect and possible mechanism of ophiopogonin D (OPD) in protecting cardiomyocytes against ophiopogonin D' (OPD')-induced injury, and provided relevant experimental data for the clinical use of Ophiopogon japonicas. Cell counting kit-8 (CCK-8) assay was used to evaluate the effect of OPD and OPD' on H9c2 cell viability. The content of reaction oxygen species (ROS) in cells were detected by flow cytometry. The contents of Fe2+ in cells were detected by FerroOrange's fluorescence imaging. The content of glutathione (GSH) and glutathione peroxidase (GSH-Px) were detected by kits. The expression of transferrin receptor 1 (TFR1), cyclooxygenase 2 (COX2), NADPH oxidase 1 (NOX1), long-chain acyl-CoA synthetase 4 (ACSL4), cationic amino acid transporter 11 (SLC7A11), glutathione peroxidase 4 (GPX4), and ferritin heavy chain 1 (FTH1) was detected by Western blot. Results showed that OPD' (1 μmol·L-1) significantly induced the expression of ferroptosis-related proteins, the contents of Fe2+, ROS, and GSH-Px were increased, and the content of GSH were decreased. In addition, different concentrations of OPD (0.5, 1, and 2 μmol·L-1) could partially reverse the myocardial cell injury caused by OPD', and the best effect was obtained when the dose range was 1-2 μmol·L-1. The experimental results show that OPD can interfere with the ferroptosis caused by OPD', and then have a protective effect on H9c2 cells.

2.
China Journal of Chinese Materia Medica ; (24): 1365-1369, 2017.
Article in Chinese | WPRIM | ID: wpr-350176

ABSTRACT

Ginsenoside Rb₁ (Rb₁), which is one of the main ingredients derived from Panax ginseng, has been found to have extensive pharmacological activities including antioxidant, anti-inflammatory, anticancer properties. In this study, the effect of Rb₁ on doxorubicin-induced myocardial autophagy was studied with H9c2 as the study object. CCK-8 method, transmission electron microscope observation, fluorescence staining observation and Western blot were used to detect changes in H9c2 cell proliferation and autophagy after treatment. According to the results, doxorubicin could cause cell viability decrease, significant increase in the LC3-Ⅱ/LC3-I ratio and down-regulation of the expression of p62. Pretreatment with ginsenoside Rb₁ inhibited cell viability decrease and increase in doxorubicin-induced autophagic structure and LC3-Ⅱ/LC3-I ratio, and down-regulation of the expression of p62. In conclusion, doxorubicin could induce H9c2 cell death and induce autophagy, and ginsenoside Rb₁ showed a protective effect on DOX-induced cardiotoxicity, which may be correlated with suppression of DOX-induced autophagy.

3.
China Journal of Chinese Materia Medica ; (24): 593-599, 2017.
Article in Chinese | WPRIM | ID: wpr-275491

ABSTRACT

To investigate the effect of clinical dose of Realgar-Indigo Naturais formula (RIF) and large-dose of Realgar on main drug-metabolizing enzymes CYP450s of rat liver, as well as its regulatory effect on mRNA expression. Wistar rats were administrated orally with tested drugs for 14 days. A Cocktail method combined with HPLC-MS/MS was used in the determination of 4 cytochrome P450 isozymes (CYP1A2, CYP2B, CYP3A and CYP2C) in liver of the rats, and the mRNA expression levels of the above subtypes were detected by real-time fluorescent quantitative PCR. The results showed that RIF can significantly induce CYP1A2 and CYP2B enzyme activity, and inhibit CYP3A enzyme activity. This result was consistent with the mRNA expression. However, its single compound showed weaker or even contrary phenomenon. Different doses of Realgar also showed significant inconsistencies on CYP450 enzymes activity and mRNA expression. These phenomena may be relevant with RIF compatibility synergies or toxicity reduction. The results can also prompt drug interactions when RIF is combined with other medicines in application.

4.
China Journal of Chinese Materia Medica ; (24): 1313-1317, 2016.
Article in Chinese | WPRIM | ID: wpr-320860

ABSTRACT

3D in vitro toxicity testing model was developed by magnetic levitation method for culture of the human hepatoma cell line HepG2 and applied to evaluate the drug hepatotoxicity. After formation of stable 3D structure for HepG2 cells, their glycogen storage capacity under 2D and 3D culture conditions were detected by immunohistochemistry technology, and the mRNA expression levels of phase Ⅰ and Ⅱ drug metabolism enzymes, drug transporters, nuclear receptors and liver-specific marker albumin(ALB) were compared between 2D and 3D culture conditions by using RT-PCR method. Immunohistochemistry results showed that HepG2 cells had abundant glycogen storage capacity under 3D culture conditions, which was similar to human liver tissues. The mRNA expression levels of major drug metabolism enzymes, drug transporters, nuclear receptors and ALB in HepG2 cells under 3D culture conditions were up-regulated as compared with 2D culture conditions. For drug hepatotoxicity evaluation, the typical hepatotoxic drug acetaminophen(APAP), and most reported drugs Polygonum multiflorum Thunb.(Chinese name He-shou-wu) and Psoraleae corylifolia L.(Chinese name Bu-gu-zhi) were selected for single dose and repeated dose(7 d) exposure. In the repeated dose exposure test, 3D HepG2 cells showed higher sensitivity. This established 3D HepG2 cells model with magnetic levitation 3D culture techniques was more close to the human liver tissues both in morphology and functions, so it was a better 3D hepatotoxicity evaluation model.

5.
China Journal of Chinese Materia Medica ; (24): 1504-1510, 2016.
Article in Chinese | WPRIM | ID: wpr-320829

ABSTRACT

To study the effect of aqueous extract of Cassiae Semen on the activity, mRNA and protein expressions of cytochrome P450(CYP450) system in rat liver microsomes, microsomes of rat liver were prepared after the oral administration with aqueous extract of Cassiae Semen for 14 days. The enzyme activity was quantified by Cocktail method. Meanwhile, the mRNA and protein expressions of CYP1A2, CYP2B1, CYP2C11, CYP2D2, CYP2E1 and CYP3A1 in the livers were detected by RT-PCR and Western blot. The result of this experiment was that aqueous extract of Cassiae Semen obviously induced the enzyme activities of CYP1A2, CYP2B1, CYP2C11, CYP2D2, CYP2E1 and CYP3A1. Low dose of aqueous extract of Cassiae Semen significantly reduced the activity of CYP2D2, but the activity of CYP2D2 was significantly induced by middle dose and high dose of aqueous extract of Cassiae Semen. These subtypes were increased in a dose-dependent manner except for CYP3A1. The mRNA levels of CYP1A2, CYP2C11, CYP2D2 and CYP2E1 were also induced in rats treated with aqueous extract of Cassiae Semen, but with no significant effect in CYP2B1 and CYP3A1 mRNA expressions. The protein levels of CYP2C11 and CYP2E1 were also induced in rats treated with aqueous extract of Cassiae Semen, but with no significant difference. Since the enzyme activity, mRNA and protein expressions of CYP450, particularly CYP2C11and2E1subtypes, were induced or inhibited by aqueous extract of Cassiae Semen to varing degrees, suggesting the potential drug-drug interactions should be concerned.

6.
China Journal of Chinese Materia Medica ; (24): 933-937, 2015.
Article in Chinese | WPRIM | ID: wpr-330207

ABSTRACT

To study the effect of Siwu decoction on the function and expression of P-glycoprotein (P-gp) in Caco-2 cells. The Real-time quantitative poly-merase chain reaction (Q-PCR) was used to analyze the mRNA expression of MDR1 gene in Caco-2 cells. Flow cytometer was used to study the effect of Siwu decoction on the uptake of Rhodamine 123 in Caco-2 cells, in order to evaluate the efflux function of P-gp. Western blotting method was used to detect the effect of Siwu decoction on the P-gp protein expression of Caco-2 cells. Compared with the blank control group, after Caco-2 incubation with Siwu decoction at concentrations of 3.3, 5.0, 10.0 g x L(-1) for 24, 48, 72 h, the mRNA expression of MDR1 was up-regulated, suggesting the effect of Siwu decoction in inducing the expression of MDR1. After the administration with Siwu decoction in Caco-2 cells for 48 h, the uptake of Rhodamine 123 in Caco-2 cells decreased by respectively 16.6%, 22.1% (P < 0.05) and 45.4% (P < 0.01), indicating that the long-term administration of Siwu decoction can enhance the P-gp efflux function of Caco-2 cells. After the incubation of Caco-2 cells with Siwu decoction for 48 h, the P-gp protein expression on Caco-2 cell emebranes, demonstrating the effect of Siwu decoction in inducing the protein expression of P-gp.


Subject(s)
Humans , ATP Binding Cassette Transporter, Subfamily B , Genetics , Metabolism , ATP Binding Cassette Transporter, Subfamily B, Member 1 , Genetics , Metabolism , Caco-2 Cells , Drugs, Chinese Herbal , Pharmacology , Up-Regulation
7.
China Journal of Chinese Materia Medica ; (24): 2737-2742, 2015.
Article in Chinese | WPRIM | ID: wpr-337898

ABSTRACT

To research the influence of Reduning injection on the activity and mRNA expression of cytochrome P450 (CYP450) system in rat liver microsomes. Rat liver microsomes were prepared after a seven-days continuous administration of Reduning injection. An HPLC-MS method was applied to determine the specific metabolites of CYP450 probe substrates in rat liver microsomal incubations. The activity of CYP450 isozymes were represented by the formation of metabolites. The Real-time quantitative polymerase chain reaction (Q-PCR) was applied to determine the mRNA expression levels of CYP450. Reduning injection significantly reduced the activity of CYP2B1, 2C12, 2C13 (P < 0.01), but did not affect CYPlA2; low dose and high dose of Reduning injection had an inhibition trend on the activity of CYP2D2, but did not statistically differ from control group; low dose of Reduning injection significantly induced the activity of CYP3A1 (P < 0.01), high dose of Reduning injection had an induce trend on the activity of CYP3A1, but did not statistically differ from control. At the mRNA level, low and high dose of Reduning injection had an induce trend on the expression of CYP1A2, 2C11, 2D1, 2E1, 3A1, but did not statistically differ from control. Reduning injection significantly induced the activity of CYP2B1. Reduning injection significantly induced the activity of CYP3A1 in mRNA expression and enzyme activity levels, which may result adverse drug reaction after being combined with macrolides antibiotics. Reduning injection significantly reduced the activity of CYP2B1, 2C12, 2C13, 2D2 in enzyme activity levels, when combined with other drugs, it should be fully taken into account of the possible drug-drug interaction in order to avoid adverse side effects.


Subject(s)
Animals , Male , Rats , Cytochrome P-450 Enzyme System , Metabolism , Drugs, Chinese Herbal , Pharmacology , Injections , Isoenzymes , Metabolism , Microsomes, Liver , Rats, Sprague-Dawley
8.
China Journal of Chinese Materia Medica ; (24): 2743-2747, 2015.
Article in Chinese | WPRIM | ID: wpr-337897

ABSTRACT

To research the effect of Ginseng Radix et Rhizoma and Aconiti Lateralis Radix Praeparata compatibility on cardiac toxicity in rats by UPLC-Q-TOF/MS, and explore the endogenous markers and molecule mechanism. Different compatibility of Shenfu decoction were given to male Wistar rats at dosage of 20 g · kg(-1) for 7 days, collected the serum, and analyze the endogenous metabolites effected by Shenfu formulation by principal component analysis and partial least-squares analysis. Results showed that content of glutathione, phosphatidylcholine and citric acid decreased in mixed-decoction group, while ascorbic acid, uric acid, D-galactose, tryptophan, L-phenylalanine increased. The results showed cardiac toxicity of Aconiti Lateralis Radix Praeparata in Shenfu mixed-decoction. Shenfu co-decoction group showed a similar or weaker trend compared with control group, but most of them do not have a statistically significant. The results indicated the scientific basis of Shenfu compatibility by comparison of co-decoction group with mixed-decoction group. Shenfu compatibility can reduce cardiac toxicity induced by Aconiti Lateralis Radix Praeparata, and citric acid, glutathione, phosphatidyl choline, uric acid might be regarded as potential markers of cardiotoxicity.


Subject(s)
Animals , Male , Rats , Biomarkers , Cardiotoxicity , Drugs, Chinese Herbal , Toxicity , Glutathione , Blood , Least-Squares Analysis , Metabolomics , Methods , Principal Component Analysis , Rats, Wistar
9.
China Journal of Chinese Materia Medica ; (24): 2748-2752, 2015.
Article in Chinese | WPRIM | ID: wpr-337896

ABSTRACT

Dioscin has a wide range of biological effects and broad application prospects. However the studies concerning the toxicology and mechanism of dioscin is small. This article is to study the hepatotoxicity of dioscin and the effect of dioscin treatment on expression of aryl hydrocarbon receptor (AhR) mRNA and CYP1A mRNA and protein in HepG2 cells in vitro. Dioscin 0.5-32 µmol · L(-1) exposed to HepG2 cells for 12 h, cell viability was examined by CCK-8 assay and the release rate of lactate dehydrogenase (LDH) was to evaluate cell membrane damage. HepG2 cells morphologic changes were quantified by inverted Microscope, and the effect on production of reactive oxygen species (ROS) was detected by flow cytometry. The mRNA expression of CYP1A and AhR was evaluated by RT-RCR. The protein expression of CYP1A1 was detected by western blot. The cell viability was significantly inhibited after HepG2 cells were exposed to dioscin 0.5-32 µmol · L(-1). Compared with the control, the LDH release rate and ROS were significantly increased. The expression of CYPlA and AhR mRNA was increased. The expression of CYP1Al protein was increased after dioscin treatment, and resveratrol, an AhR antagonist, could downregulate the expression of CYP1A1. It follows that large doses dioscin has potential hepatotoxicity. The possible mechanism may be dioscin can active aryl hydrocarbon receptor (AhR) and induce the expression of CYP1A.


Subject(s)
Humans , Cell Survival , Chemical and Drug Induced Liver Injury , Cytochrome P-450 CYP1A1 , Genetics , Diosgenin , Toxicity , Hep G2 Cells , L-Lactate Dehydrogenase , Bodily Secretions , RNA, Messenger , Reactive Oxygen Species , Metabolism , Receptors, Aryl Hydrocarbon , Genetics
10.
China Journal of Chinese Materia Medica ; (24): 3444-3449, 2015.
Article in Chinese | WPRIM | ID: wpr-237691

ABSTRACT

Pregnane X receptor (PXR) is key transcription factors which mainly regulate the expression of CYP3A genes. At the molecular level, PXR has been revealed the protection mechanism of the body against xenochemicals and a major mode of the drug-drug interactions. Besides playing an important role in drug metabolism and interactions, PXR and its target genes also play an important role in maintaining normal physiological function and homeostasis. Therefore, it is necessary to study the regulation of PXR and its related pharmacological effects of TCM and natural products, and to provide new clues for the new pharmacological pathway.


Subject(s)
Animals , Humans , Drug Evaluation, Preclinical , Drugs, Chinese Herbal , Pharmacology , Gene Expression , Receptors, Steroid , Genetics , Metabolism
11.
Chinese Journal of Natural Medicines (English Ed.) ; (6): 730-737, 2014.
Article in English | WPRIM | ID: wpr-812207

ABSTRACT

AIM@#Radiation induces an important apoptosis response in irradiated organs. The objective of this study was to investigate the radioprotective effect of tetramethylpyrazine (TMP) on irradiated lymphocytes and discover the possible mechanism of protection.@*METHOD@#Lymphocytes were pretreated for 12 h with TMP (25-200 μmol·L(-1)) and then exposed to 4 Gy radiation. Cell apoptosis and the signaling pathway were analyzed.@*RESULTS@#Irradiation increased cell death, DNA fragmentation, activated caspase activation and cytochrome c translocation, downregulated B-cell lymphoma 2 (Bcl-2) and up-regulated Bcl-2-associated X protein (Bax). Pretreated with TMP significantly reversed this tendency. Several anti-apoptotic characteristics of TMP, including the ability to increase cell viability, inhibit caspase-9 activation, and upregulate Bcl-2 and down-regulate Bax in 4Gy-irradiated lymphocytes were determined. Signal pathway analysis showed TMP could translate nuclear factor-κB (NF-κB) from cytosol into the nucleus.@*CONCLUSION@#The results suggest that TMP had a radioprotective effect through the NF-κB pathway to inhibit apoptosis, and it may be an effective candidate for treating radiation diseases associated with cell apoptosis.


Subject(s)
Humans , Apoptosis , Radiation Effects , Cell Line , Cell Survival , Radiation Effects , DNA Fragmentation , Radiation Effects , Drugs, Chinese Herbal , Pharmacology , Lymphocytes , Cell Biology , Radiation Effects , NF-kappa B , Genetics , Metabolism , Proto-Oncogene Proteins c-bcl-2 , Genetics , Metabolism , Pyrazines , Pharmacology , Radiation-Protective Agents , Pharmacology , bcl-2-Associated X Protein , Genetics , Metabolism
12.
Chinese Journal of Integrated Traditional and Western Medicine ; (12): 606-610, 2014.
Article in Chinese | WPRIM | ID: wpr-312768

ABSTRACT

<p><b>OBJECTIVE</b>To screen active components in Compound Danshen (CD) based on pregnane X receptor-cytochrome P450 3A4 (PXR-CYP3A4).</p><p><b>METHODS</b>By using PXR-CYP3A stable transfection human hepatoblastoma G2 (HepG2) cell lines engineering cell strain combined reporter genes technology, active components that induce or inhibit PXR-CYP3A4 paths in CD were screened, and confirmed at the level of enzymic activities. The experiment was divided into the positive control group (RIF 10 micro mol/L), the DMSO group (DMSO 0.1%), each dose of treatment groups (ginsenoside Rc, Rf, Rb2, Rg2, F2, F1, tanshinone I , isoborneol 5, 10, 25, 50, 100, and 200 micro mol/L; each with six duplicates). Cells medium was removed 36, 48, and 60 h after treatment. The activity of CYP3A4 was then determined in the supernant and the fold induction was calculated.</p><p><b>RESULTS</b>Compared with the DMSO group, the fold induction increased when ginsenoside Rc, Rf, Rb2, Rg2, F2, F1, tanshinone I , and isoborneol 50 and 100 micro mol/L was respectively intervened for 36, 48, and 60 h (P <0.05). When cells were treated with isoborneol 200 micro mol/L for 48 and 60 h,the fold induction of ginsenoside Rb2, Rg2, and F1 was significantly higher than that of the RIF group (P <0.05). Enzymic activity results showed that ginsenoside Rc, Rf, Rb2, F2, and F1 could increase the enzyme activity of CYP3A4 at 48 h (P <0.05).</p><p><b>CONCLUSION</b>Ginsenoside Rc, Rf, Rb2, F2, F1, tanshinone I, and isoborneol in DC could induce CYP3A4 enzymes.</p>


Subject(s)
Humans , Cytochrome P-450 CYP3A , Metabolism , Abietanes , Drugs, Chinese Herbal , Chemistry , Genes, Reporter , Ginsenosides , Metabolism , Hep G2 Cells , Receptors, Steroid , Metabolism , Salvia miltiorrhiza , Transfection
13.
China Journal of Chinese Materia Medica ; (24): 3824-3828, 2014.
Article in Chinese | WPRIM | ID: wpr-310981

ABSTRACT

To study the effect of Panax notoginseng saponins (PNS) on liver drug metabolic enzyme activity, mRNA and protein expressions in rats. Male Wistar rats were randomly divided into nine groups. After administration of the test drugs, their liver microsomes, liver total RNA and total protein were extracted to detect the regulating effect of PNS on liver drug metabolic enzyme activity-related subtype enzymatic activity, mRNA and protein expression by substrate probe, quantitative PCR and Western Blot technology. The result of this experiment was that PNS could significantly induce CYP1A2 and CYP2E1 enzyme activity, mRNA expression, CYP2E1 protein expression level. PNS significantly induced CYP3A mRNA expression, but with no significant effect in CYP3A enzyme activity level. PNS had no significant effect CYP1A1 and CYP2B mRNA expressions and enzyme activity levels. PNS had selective regulations on different P450 subtypes, and the major subtypes were CYP1A2 and CYP2E1. In clinical practice, particularly in the combination with CYP1A2 and CYP2E1 metabolism-related drugs, full consideration shall be given to the possible drug interactions in order to avoid potential toxic and side effects. Meanwhile, whether the induction effect of CYP2E1 gets involved in ginsenoside's effect incavenging free radicals deserves further studies.


Subject(s)
Animals , Male , Cytochrome P-450 Enzyme System , Genetics , Metabolism , Drugs, Chinese Herbal , Pharmacology , Liver , Microsomes, Liver , Panax notoginseng , Chemistry , Rats, Wistar , Saponins , Pharmacology
14.
China Journal of Chinese Materia Medica ; (24): 3720-3725, 2013.
Article in Chinese | WPRIM | ID: wpr-291296

ABSTRACT

To study the effect of Siwu decoction (SWD) compound and its combined administration on hepatic P450 enzymatic activity and mRNA expression in rats. Rats were orally administered with SWD and water decoction combined with other medicines for two weeks, and then sacrificed. Their livers were perfused with normal saline to prepare liver micrisomes. Mixed probe and liver microsome in vitro incubation method were adopted to detect the effect of SWD on hepatic cytochrome P450. The real-time quantitative polymerase chain reaction (Q-PCR) was used to detect the effect of SWD on the expression of hepatic cytochrome P450. Compared with the control group, the SWD compound group showed higher CYP1A2 enzymatic activity (P < 0.05); Rehmanniae-paeoniae, angelicae-paeoniae, angelicae-rhizome, paeoniae-rhizome groups had lower CYP1A2 and CYP2C19 enzymatic activities (P < 0.05); And the compound group, the single component group and the combination group showed lower CYP2B6 enzymatic activities (P < 0.05). The compound could up-regulated the mRNA expression of CYP2B1 (P < 0.05); And the four single components could down-regulated the mRNA expression of CYP2B1 (P < 0.05). SWD compound had the effect in inducing CYP1A2 enzymatic activity. The rehmanniae-paeoniae group and the angelicae-paeoniae group had identical enzymatic activity with the control group, but significant down-regulation in CYP1A2 enzymatic activity after being combined with paeoniae. The compound and its combined administration showed the inhibitory effect on CYP2B6 enzymatic activity, particularly being combined with angelicae. The compound showed identical effect with the four single components in terms of CYP1A2 mRNA expression and enzymatic activity.


Subject(s)
Animals , Rats , Cytochrome P-450 Enzyme System , Genetics , Metabolism , Drugs, Chinese Herbal , Pharmacology , Liver , Microsomes, Liver , RNA, Messenger , Genetics , Metabolism , Rats, Wistar
15.
Chinese Journal of Integrated Traditional and Western Medicine ; (12): 686-690, 2013.
Article in Chinese | WPRIM | ID: wpr-287487

ABSTRACT

<p><b>OBJECTIVE</b>To study the toxicity changes of different proportions of Radix Adenophora, Radix Glehniae combined with Veratrum nigrum L., thus providing acute toxicity data and investigating whether decoction factors were correlated with toxicity.</p><p><b>METHODS</b>The uniform design method was used by two factors and seven levels to investigate the toxicity changes in different proportions of Radix Adenophora, Radix Glehniae combined with Veratrum nigrum L. The decoction factors were also investigated.</p><p><b>RESULTS</b>The compatibility toxicity was affected mainly by Veratrum nigrum L. and the toxicity increased along with increased doses of Veratrum nigrum L. The toxicity of co-decoction was higher than mixed decoction in the same dosage of Radix Glehniae and Veratrum nigrum L. The promotion of the dissolution of the toxic component of Veratrum nigrum L. in co-decoction may be the cause of the higher toxicity.</p><p><b>CONCLUSION</b>Radix Adenophora and Radix Glehniae combined with Veratrum nigrum L. resulted in higher toxicity, which indicated that the incompatibility between Radix Adenophora, Radix Glehniae, and Veratrum nigrum L. In clinic practice, a prescription contained these drugs should be avoided.</p>


Subject(s)
Animals , Female , Male , Mice , Drug Antagonism , Drug Incompatibility , Drugs, Chinese Herbal , Toxicity
16.
Journal of Integrative Medicine ; (12): 327-336, 2013.
Article in English | WPRIM | ID: wpr-308237

ABSTRACT

<p><b>OBJECTIVE</b>To evaluate whether Shenfu injection (SFI) protects against cardiac myocyte injury induced by Fupian injection (FPI) in vitro.</p><p><b>METHODS</b>H9c2 cells were separately treated with FPI, Renshen injection (RSI) and SFI. Cell viability, lactate dehydrogenase (LDH) release, spontaneous beating rate of primative cardical cells, caspase-3/7 activity, cell apoptosis, and cytochrome P450 2J3 (CYP2J3) mRNA expression were analyzed.</p><p><b>RESULTS</b>The viability of H9c2 cells treated with SFI (37 and 75 mg/mL) was significantly higher than that of H9c2 cells treated with FPI (25 and 50 mg/mL) (P<0.05, P<0.01, respectively). LDH activity of H9c2 cells treated with SFI (75 mg/mL) was significantly decreased (P<0.01) compared with that of H9c2 cells treated with FPI (50 mg/mL). SFI (150 mg/mL) significantly attenuated FPI (100 mg/mL)-induced spontaneous beating rate decrease in primary myocardial cells after 4-hour treatment. Compared with FPI (12 and 25 mg/mL), SFI (18 and 37 mg/mL) treatment could effectively reverse the change of caspase-3/7 activity (P<0.01 and P<0.01, respectively). Compared with FPI (6 and 25 mg/mL), apoptotic cells decreased significantly (P<0.05, P<0.01, respectively) when H9c2 cells were incubated with SFI (9 and 37 mg/mL). The expression of CYP2J3 mRNA was down-regulated by FPI, while RSI and SFI could up-regulate the expression of CYP2J3 (P<0.01), which suggested the potential mechanism of protection of RSI against cardiac myocyte damage induced by FPI treatment.</p><p><b>CONCLUSION</b>These observations indicate that SFI has the potential to exert cardioprotective effects against FPI toxicity. The effect was possibly correlated with the activation of CYP2J3.</p>


Subject(s)
Animals , Rats , Apoptosis , Caspases , Metabolism , Cell Survival , Cells, Cultured , Cytochrome P-450 Enzyme System , Genetics , Physiology , Drugs, Chinese Herbal , Pharmacology , L-Lactate Dehydrogenase , Bodily Secretions , Myocytes, Cardiac
17.
Acta Pharmaceutica Sinica ; (12): 144-148, 2013.
Article in Chinese | WPRIM | ID: wpr-235690

ABSTRACT

In order to study effects of ginseng on the metabolism of drug belong to CYP3A4 substrate, screening of pregnane X receptor activation from ginsenosides was performed by reporter assay. Based on PXR-CYP3A stable translation cell lines, 13 ginsenosides were screened for pregnane X receptor activation by reporter assays, and RIF as the positive control. The effect of ginsenosides Rg1 onCYP3A4 mRNA expression was also investigated by RT-PCR. The PXR-CYP3A stable translation cell lines had good response to RIF, and the EC50 is 2.51 micro mol x L(-1). When the condition of final concentration was 10 micromol x L(-1), ginsenoside F2 and protopanaxatriol had moderate inductive effects on PXR. Panaxotriol, Rg2, pseudoginsenoside F11, Rg1, ginsenoside and Rb3 had inhibitory effects on PXR. Ginsenoside Rf1, Rg3, Rh2 and protopanaxdiol had no obvious effects on PXR. Rg1 down-regulated CYP3A4 mRNA expression in a concentration-dependent manner. Activation of pregnane X receptor by ginsenosides may influence the metabolism of drug belong to CYP3A4 substrate, and cause ginseng-drug interactions.


Subject(s)
Humans , Cytochrome P-450 CYP3A , Genetics , Metabolism , Drug Interactions , Ginsenosides , Pharmacology , Hep G2 Cells , RNA, Messenger , Metabolism , Receptors, Steroid , Genetics , Sapogenins , Pharmacology , Transfection
18.
Acta Pharmaceutica Sinica ; (12): 728-733, 2013.
Article in Chinese | WPRIM | ID: wpr-235603

ABSTRACT

The paper is to report the study of the effect of Shenfu injection on the enzyme activity of liver CYP450 and its mRNA level of rat liver. Microsome of rat liver was prepared after intravenous administration of Shenfu injection for 7 days. The enzyme activity was quantified by Cocktail method. Meanwhile, the mRNA expression of CYP1A2, CYP2B1/2, CYP2C11 and CYP3A1 in the liver was detected by RT-PCR. Shenfu injection obviously induced the enzyme activities of CYP2B and CYP2C. Meantime Shenfu injection decreased the enzyme activities of CYP1A2 and CYP3A. The mRNA levels of CYP2B and CYP2C were also induced in rats treated with Shenfu injection. But it obviously inhibited the mRNA level of CYP1A2 and CYP3A. Since the enzyme activity and mRNA level were obviously changed after administration, the potential effect of drug-drug interaction should be concerned.


Subject(s)
Animals , Male , Rats , Aconitum , Chemistry , Aryl Hydrocarbon Hydroxylases , Genetics , Metabolism , Cytochrome P-450 CYP1A2 , Genetics , Metabolism , Cytochrome P-450 CYP2B1 , Genetics , Metabolism , Cytochrome P-450 CYP3A , Genetics , Metabolism , Cytochrome P-450 Enzyme System , Genetics , Metabolism , Cytochrome P450 Family 2 , Drug Combinations , Drugs, Chinese Herbal , Pharmacology , Injections , Microsomes, Liver , Panax , Chemistry , Plants, Medicinal , Chemistry , RNA, Messenger , Metabolism , Rats, Sprague-Dawley , Steroid 16-alpha-Hydroxylase , Genetics , Metabolism
19.
Acta Pharmaceutica Sinica ; (12): 1488-1492, 2011.
Article in Chinese | WPRIM | ID: wpr-323096

ABSTRACT

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.


Subject(s)
Aconitine , Aconitum , Chemistry , Alkaloids , Chromatography, High Pressure Liquid , Methods , Drug Combinations , Drugs, Chinese Herbal , Chemistry , Panax , Chemistry , Plants, Medicinal , Chemistry , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization , Methods
20.
China Journal of Chinese Materia Medica ; (24): 420-423, 2008.
Article in Chinese | WPRIM | ID: wpr-284468

ABSTRACT

<p><b>OBJECTIVE</b>To research the effects of Siwu tang on serum protein of blood deficiency using proteomic technique and further explore its potential molecular mechanism to cure blood deficiency.</p><p><b>METHOD</b>The sera of normal, blood deficiency and cured group were collected. Proteomic protocol involving the high resolution two-dimensional polyacryamide gel electrophoresis, the computer-assisted image analysis, and the mass spectrometry was used to detect regulated protein by Siwu tang.</p><p><b>RESULT</b>Compared with normal group, there were 15 proteins changed, in which 11 increased and 4 decreased expressed proteins in sera could be recovered by Siwu tang. The up-regulated proteins involved haptoglobin, clusterin, complement component C4B and GTP binding protein 2, while the down-regulated proteins involved transthyretin and heamoglobin beta.</p><p><b>CONCLUSION</b>Siwu tang could regulate serum protein, which include immunology, apoptosis, DNA injury repair, and blood ingredients. This might be the mechanism of Siwu tang to cure blood deficiency.</p>


Subject(s)
Humans , Blood Proteins , Drugs, Chinese Herbal , Pharmacology , Hematologic Diseases , Blood , Drug Therapy , Proteomics , Methods
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