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The occurrence of benign prostate hyperplasia(BPH)was related to disrupted sex steroid hormones,and metformin(Met)had a clinical response to sex steroid hormone-related gynaecological disease.How-ever,whether Met exerts an antiproliferative effect on BPH via sex steroid hormones remains unclear.Here,our clinical study showed that along with prostatic epithelial cell(PEC)proliferation,sex steroid hormones were dysregulated in the serum and prostate of BPH patients.As the major contributor to dysregulated sex steroid hormones,elevated dihydrotestosterone(DHT)had a significant positive rela-tionship with the clinical characteristics of BPH patients.Activation of adenosine 5'-monophosphate(AMP)-activated protein kinase(AMPK)by Met restored dysregulated sex steroid hormone homeostasis and exerted antiproliferative effects against DHT-induced proliferation by inhibiting the formation of androgen receptor(AR)-mediated Yes-associated protein(YAP1)-TEA domain transcription factor(TEAD4)heterodimers.Met's anti-proliferative effects were blocked by AMPK inhibitor or YAP1 over-expression in DHT-cultured BPH-1 cells.Our findings indicated that Met would be a promising clinical therapeutic approach for BPH by inhibiting dysregulated steroid hormone-induced PEC proliferation.
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Total glucosides of Rhizoma Smilacis Glabrae (RSG) are selective immunosuppressants that exhibit primary efficacy in the treatment of rheumatoid arthritis through targeted inhibition of activated T cells. In this study, we aimed to investigate the potential application of RSG in the treatment of psoriasis and elucidate its mechanism of action and material basis. Our findings revealed significant improvements upon administration of RSG in an imiquimod (IMQ)-induced psoriasis model. These improvements were characterized by a remarkable increase in the number of tail scales in mice and a substantial amelioration of skin erythema, ulceration, and flaking. By transcriptome sequencing and T-cell flow sorting assay, we identified notable effects of RSG on the modulation of various cellular processes. Specifically, RSG prominently down-regulated the Th17/Treg ratio in damaged skin tissues and reduced the proportion of G2 phase cells. Furthermore, RSG exhibited a stimulatory effect on the proliferation and differentiation of epithelial cells. Of particular interest, we discovered that β-sitosterol, sitostenone, stigmasterol, smiglanin, and cinchonain Ib displayed potent inhibitory effects on the IL-17-mediated inflammatory response in HaCaT cells. In summary, our study highlights the therapeutic potential of RSG in the treatment of psoriasis, attributed to its ability to regulate the Th17/Treg balance. These findings contribute to the development of new indications for RSG and provide a solid theoretical foundation for further exploration in this field.
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Animaux , Souris , Lymphocytes T régulateurs , Psoriasis/traitement médicamenteux , Polyarthrite rhumatoïde , Dosage biologique , Glucosides/pharmacologieRésumé
ObjectiveTo establish a more accurate and sensitive liquid chromatography-ultraviolet (LC-UV) method for the determination of uric acid in rat serum, and compare the results with those of commercial kits, providing a new method for the accurate determination of uric acid in the rat hyperuricemia model induced by potassium oxonate.Methods A hyperuricemia model was established by intraperitoneal injection of potassium oxonate (300 mg/kg) into SPF-grade male SD rats, and the control group was administered an equal amount of 0.5% sodium carboxymethylcellulose solution. Blood samples were collected from the posterior orbital venous plexus and centrifuged to obtain serum samples. After precipitation with 0.1% trifluoroacetic acid-acetonitrile (containing the internal standard 3,4-dihydroxybenzylamine hydrobromide), the supernatant was injected for analysis. Uric acid was separated on a Waters XBridge HILIC column (150 mm×4.6 mm, 3.5 μm) using acetonitrile (containing 0.5% formic acid and 2 mmol/mL ammonium formate) as the organic phase and methanol solution (methanol∶water=1∶1, containing 0.5% formic acid with 2 mmol/L ammonium formate) as the aqueous phase for isocratic elution and detection at 290 nm. Serum samples treated with activated carbon were used as substitute matrices for the methodological verification. Serum uric acid levels in rats with potassium oxonate-induced hyperuricemia were measured using the established LC-UV method and commercially available kits (uricase and phosphotungstic acid methods), and the accuracies of the three methods were compared.Results Serum uric acid showed a good linear relationship (R>0.999) at mass concentration of 10–200 μg/mL in rats, the lower limit of quantification was 10 μg/mL, the accuracy ranged from -2.17% to 2.21%, the intra-batch precision ranged from 0.52% to 1.95%, the inter-batch precision ranged from 3.04% to 4.90%, and the extraction recovery ranged from 83.12% to 89.91%. In the rat model, the results obtained using the commercially available phosphotungstic acid method kit were significantly higher than those of the LC-UV method, and those obtained using the commercially available uricase method kit were significantly lower than those of the LC-UV method, but the LC-UV method showed the best recovery of the spiked sample (95.90%–99.96%).ConclusionThe LC-UV method developed in this study can determine the concentration of uric acid in rat serum with higher accuracy than commercially available kits and is recommended for the determination of serum uric acid in the rat model of hyperuricemia induced by potassium oxonate.
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Cholesterol is an important lipid component in the body, which not only participates in the formation of cell membranes, but also is the raw material for the synthesis of bile acids and steroid hormones. Low density lipoprotein receptor (LDLR) is involved in cholesterol metabolism and plays an important role in maintaining the cholesterol homeostasis of organism cells. The expression of LDLR is precisely regulated by transcription, post-transcription and post-translation, and the imbalance of ldlr expression will lead to the occurrence and development of many diseases. In this paper, the molecular regulation mechanism of LDLR, the damage of target organs caused by the imbalance of LDLR expression and the research and development progress of drugs targeting LDLR are reviewed, which provides theoretical basis for further understanding of the progress of diseases related to lipid metabolism disorder and new insights for developing drugs targeting LDLR with more effective and less side effects.
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Estrogen receptors are steroid receptors, widely distributed in skeletal muscle, liver and other tissues.Currently, there are 5 known estrogen receptors.Different estrogen receptors are distributed in different places and have different functions.By mediating different estrogen receptors, estrogen plays an important role in anti-inflammation, promoting proliferation and inhibiting muscle atrophy.Skeletal muscle is the main tissue in the human body, accounting for about 40% of body weight.Skeletal muscle not only plays the role of exercise and support, but also plays an important role in maintaining the body′s metabolism.In recent years, estrogen receptors have received extensive attention in skeletal muscle diseases.Estrogen receptors are considered as potential targets for the treatment of skeletal muscle diseases such as Duchenne muscular dystrophy(DMD)and myotubular myopathy(MTM). This article reviews the research progress of estrogen receptors in skeletal muscle diseases.
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@#Duchene muscular dystrophy (DMD) is a serious progressive muscular dystrophy.Reports in recent years about abnormal lipid in DMD patients have increased, yet little attention has been paid to liver lipid.This study aimed to explore the effect of dystrophin gene defect on liver lipid synthesis.7-week-old mdx male mice were used as DMD model.The conditions of liver function, liver lipid accumulation and liver lipid synthesis were determined through liver tissue morphological examination, blood biochemical examination, and detection of hepatic gene and protein expression.The results showed that lipid droplets in liver of mdx mice increased significantly.The contents of total cholesterol and triglyceride in liver, aspartate aminotransferase and alanine aminotransferase in serum increased.The gene and protein expression of hepatic lipid synthesis-related enzymes such as fatty acid synthase, acetyl CoA carboxylase, and sterol regulatory element binding protein 1-c were up-regulated.These results showed accumulation of liver lipid in 7-week-old mdx male mice.
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@#To investigate the ameliorative effect of psoralen (PSO) on carbon tetrachloride (CCl4)-induced acute liver injury in mice and its potential mechanism, female C57BL/6J mice aged 6-8 weeks were continuously administrated with psoralen or positive control drug diallyl sulfide (DAS) intragastrically for 4 days.On day 4, except that the control group were treated with vehicle control, other groups were all given carbon tetrachloride intraperitoneally to establish a carbon tetrachloride acute liver injury model.Serum biochemical indicators alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels were detected; liver pathological changes were observed by HE staining; cytochrome P450 2E1 (CYP2E1) protein levels were detected by Western blot; the protein level of CYP2E1 were detected by immunohistochemistry (IHC) staining; and the gene levels of CYP2E1, tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) were detected by RT-PCR.Compared with the model group, psoralen could improve the inflammatory cell infiltration and hepatocyte necrosis caused by carbon tetrachloride, significantly reducing the serum ALT and AST levels, down-regulating the inflammatory factors TNF-α and IL-6 levels, and inhibiting CYP2E1 protein expression.The results show that psoralen can ameliorate the acute liver injury induced by carbon tetrachloride in mice, with the possible mechanism inhibiting the protein expression of CYP2E1.
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Previously, we proposed a new perspective of triptolide (TP)-associated hepatotoxicity: liver hypersensitivity upon lipopolysaccharide (LPS) stimulation. However, the mechanisms for TP/LPS-induced hepatotoxicity remained elusive. The present study aimed to clarify the role of LPS in TP/LPS-induced hepatotoxicity and the mechanism by which TP induces liver hypersensitivity upon LPS stimulation. TNF- inhibitor, etanercept, was injected intraperitoneally into mice to investigate whether induction of TNF- by LPS participated in the liver injury induced by TP/LPS co-treatment. Mice and hepatocytes pretreated with TP were stimulated with recombinant TNF- to assess the function of TNF- in TP/LPS co-treatment. Additionally, time-dependent NF-B activation and NF-B-mediated pro-survival signals were measured and . Finally, overexpression of cellular FLICE-inhibitory protein (FLIP), the most potent NF-B-mediated pro-survival protein, was measured and to assess its function in TP/LPS-induced hepatotoxicity. Etanercept counteracted the toxic reactions induced by TP/LPS. TP-treatment sensitized mice and hepatocytes to TNF-, revealing the role of TNF- in TP/LPS-induced hepatotoxicity. Mechanistic studies revealed that TP inhibited NF-B dependent pro-survival signals, especially FLIP, induced by LPS/TNF-. Moreover, overexpression of FLIP alleviated TP/LPS-induced hepatotoxicity and TP/TNF--induced apoptosis . Mice and hepatocytes treated with TP were sensitive to TNF-, which was released from LPS-stimulated immune cells. These and other results show that the TP-induced inhibition of NF-B-dependent transcriptional activity and FLIP production are responsible for liver hypersensitivity.
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@#The occurrence of cholestatic liver injury is accompanied by the alterations of hepatocyte polarization and bile acid homeostasis. Located in epithelial cells, tight junctions(TJs)are a special barrier structure which are important in maintaining permeability and bile acid homeostasis. Based on the fully analysis and discussion of TJs, the latest therapeutic drugs for cholestasis were summaried, which may provide new perspectives and potential therapeutic agents for the treatment of cholestatic liver injury.
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To observe the effects of emodin ( Emo) on acute fatty liver in mice induced by DL-ethionine ( DL-Eth) or tetracyclin ( Tetra) and its potential mechanism, ICR mice of acute fatty liver model induced by DL-Eth were orally administered with Emo or positive control, ursodeoxycholic acid ( UDCA) for 7 days. On day 7, except that the control and Emo groups were treated with vehicle control, animals were orally administered with DL-Eth to induce acute fatty liver model. ICR mice of acute fatty liver model induced by Tetra were orally administered with Emo or positive control, Dong Bao Gan Tai ( DB) or total flavonoid C-glycosides from Abrus mollis extract ( AME) for 7 days. From day 4, except that the control group was treated with vehicle control, animals were injec-ted with Tetra intraperitoneally for 4 days to induce acute fatty liver model. Liver histopathological analyses were determined by HE staining. Serum aspartate transaminase ( AST) , alanine transaminase ( ALT) , serum triglyceride ( TG) , hepatic TG and hepatic total cholesteol ( TC) were detected. The expression of phosphorylated AMP-activa-ted kinase ( p-AMPK) , phosphorylated acetyl-CoA carboxylase ( p-ACC) , SREBP1 and fatty acid synthase ( FAS) were determined by Western blot. The expression of fatty acid translocase ( CD36 ) , peroxisome proliferator activa-ted receptor alpha ( PPARα) and microsomal triglyceride transfer protein ( MTTP ) in liver were determined by RT-PCR. Compared with model groups, Emo could improve hepatocyte swelling and hepatic steatosis induced by DL-Eth or Tetra. Serum AST, ALT, serum TG, hepatic TG and hepatic TC were decreased by Emo significantly. DL-Eth-induced increase of fatty acid synthetase-associated protein was down-regulated by Emo. Fatty acid uptake was down-regulated by Emo; fatty acid oxidation and secretion were increased by Emo. Emo might be effective in preventing acute fatty liver in mice induced by DL-Eth or Tetra.
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Aim To investigate the hypoglycemic pathway of terpenes from Cornus officinalis(TCF) from three aspects of insulin dependence, α-glucosidase inhibition, insulin sensitizing.Methods Insulin-deficient diabetes mellitus(DM) model was induced by tail vein injection of streptozotocin(STZ) into SD rats at the dose of 50mg·kg-1 body weight.Rats were randomly divided into seven groups: control group(CON), model group(Model), metformin group(Met) 0.1g·kg-1, shenqi jiangtang granules(Shenqi) group 1.0 g·kg-1, three dose groups of TCF: 0.10, 0.05, 0.025 g·kg-1.Body weight and blood glucose were measured every week.After four weeks, glycosylated hemoglobin (HbA1c), glycosylated serum protein (GSP) were determined.Normal ICR mice were divided into seven groups: CON, Model, Met group 0.2g·kg-1, acarbose group(Acar) 0.1 g·kg-1, Shenqi group 1.5g·kg-1, three dose groups of TCF: 0.20g·kg-1;0.10g·kg-1;0.05 g·kg-1.After 10 days of administration, intraperitoneal injections of glucose and gavage starch tolerance tests were employed.Normal SD rats were divided into six groups: CON, rosiglitazone group 0.02 g·kg-1, glipizide group 0.02 g·kg-1, three dose groups of TCF: 0.10, 0.05, 0.025 g·kg-1.After seven days of administration, intraperitoneal glucose tolerance test(IPGTT) was employed and levels of insulin was determined.Results (1)High dose of TCF significantly reduced the level of HbA1c(P<0.05), GSP(P<0.05) on STZ model rats;(2)TCF significantly improved the glucose tolerance and gavage starch tolerance in ICR mice(P<0.05);(3) High dose of TCF significantly reduced the blood glucose and serum insulin level.Conclusions TCF has obvious effects on inhibiting glucose absorb and promoting the use of glucose.It is able to exert hypoglycemic effect through non-insulin dependent pathway, whereas, whether it has the effects of α-glucosidase inhibition and insulin sensitization should be further validated.
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The clinical electroencephalogram (EEG) monitoring systems based on personal computer system can not meet the requirements of portability and home usage. The epilepsy patients have to be monitored in hospital for an extended period of time, which imposes a heavy burden on hospitals. In the present study, we designed a portable 16-lead networked monitoring system based on the Android smart phone. The system uses some technologies including the active electrode, the WiFi wireless transmission, the multi-scale permutation entropy (MPE) algorithm, the back-propagation (BP) neural network algorithm, etc. Moreover, the software of Android mobile application can realize the processing and analysis of EEG data, the display of EEG waveform and the alarm of epileptic seizure. The system has been tested on the mobile phones with Android 2. 3 operating system or higher version and the results showed that this software ran accurately and steadily in the detection of epileptic seizure. In conclusion, this paper provides a portable and reliable solution for epileptic seizure monitoring in clinical and home applications.
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Humains , Algorithmes , Téléphones portables , Électrocardiographie , Électroencéphalographie , Entropie , Épilepsie , Diagnostic , Monitorage physiologique , , LogicielRésumé
@#To investigate the hypoglycemic effects of terpenes from Fructus Corni(TFC)on type 2 diabetes mellitus, the db/db diabetic mice were intragastrically administered with 25, 50, 100 mg/kg of TFC for 10 weeks. The fasting blood glucose, insulin(Ins), glycosylated serum protein(GSP), total cholesterol(TC)and triglyceride(TG)levels were determined. At weeks 8 and 10, intraperitoneal injections of glucose and gavage starch tolerance tests were performed, respectively. The db/db mice showed obvious obesity. Each dose of TFC could significantly reduce the body weight of db/db mice(P< 0. 05). After 4 weeks of administration, all doses of TFC significantly reduced the fasting blood glucose of db/db mice(P< 0. 05). The serum TC, TG levels were also significantly decreased in the TFC middle- and high-dose groups(P< 0. 05). In addition, middle- and high-dose of TFC could significantly reduce the level of GSP. Middle- and high-dose of TFC also significantly improve the glucose tolerance and gavage starch tolerance in db/db mice(P< 0. 05). These results suggest that TFC could improve diabetes-related symptoms via regulating glucose and lipids metabolism.
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@#Sphingosine 1-phosphate(S1P)is a pleiotropic sphingolipid metabolite that has been shown to regulate important physiological function by activation of its G-protein-coupled receptors S1PRs. S1P has been identified as an important signaling molecule in maintaining epithelial and endothelial barrier function. S1P signaling pathway is involved in epithelial and endothelial barrier function by regulation of adherens junction and tight junction assembly, cytoskeletal reorganization, and focal adhesion formation. Thus, S1P signaling pathway may become a novel therapeutic target for cell barrier dysfunction during some illnesses such as acute lung injury, inflammatory bowel disease and sepsis. In this review, the research progress of S1P signaling pathway in regulating epithelial and endothelial barrier function and the application of S1P in barrier dysfunction-related diseases were summarized, so as to provide references for future research.
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@#A qualitative analytical method of liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry(HPLC-QTOF-MS)was developed for identification of major constituents in process residue of tripterygium glycosides. The HPLC-QTOF-MS assay was performed on a Zorbax SB-C18 column(4. 6 mm × 50 mm, 1. 8 μm)with the mobile phase consisting of acetonitrile and water containing 0. 2% formic acid in gradient mode. Positive ion mode was used for TOF-MS. According to the accurate molecular weight, MS fragment pathway, comparison with the retention time of reference compounds, total 30 compounds, including fifteen alkaloids, ten diterpenoids, four triterpenoids and an unsaturated fatty acid were identified or tentatively characterized in process residue of tripterygium glycosides. This study may be helpful to the comprehensive exploitation and utilization of process residue of tripterygium glycosides.
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<p><b>OBJEVTIVE</b>To synthesize phenoxybutyric acid derivatives as 5α-reductase inhibitors and test their biological activities in vitro.</p><p><b>METHODS</b>Eight analogues as nonsteroidal 5α-reductase inhibitors were designed and synthesized by substitution reaction of 6-(4-phenyl-piperazine-1-yl)-3(2H)-pyridazinone with phenoxybutyric acid derivatives.</p><p><b>RESULTS AND CONCLUSION</b>The structures of the compounds were characterized by 1H-NMR and MS. Biological evaluation indicated that 7 out of the 8 compounds exhibited moderate 5α-reductase inhibitory activities, especially the compounds A1 and A7 with inhibition rates reaching 12.50% and 19.64% at the concentration of 3.3 × 10⁻⁵ mol/L, respectively.</p>
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Inhibiteurs de la 5-alpha réductase , Pharmacologie , Butyrates , Chimie , Pharmacologie , Conception de médicamentRésumé
<p><b>OBJECTIVE</b>To observe the liver injury degree of SD rats after 28-day administration of aqueous extracts of Polygonum multiflorum (AEPM) and the correlation with cholestasis mechanism.</p><p><b>METHOD</b>Adult SD rats were orally administered with 30, 60 g x kg(-1) of APEM once every day for 28 d. After 28 d, the general condition of rats such as weight were observed, liver function-related indicators were detected. Bile was collected to determine total bile acid output, flow rate and density and changes in major compositions. Their livers were weighed then sent for histopathological examination.</p><p><b>RESULT</b>AEPM did not change the general conditions and weights of rats. From the results of the related indicators of liver function and cholestasis, AEPM did not change the contents of ALT and AST in serum, but high dose of AEPM can increase the contents of ALP, GGT and TBA in serum (P < 0.05, P < 0.01) and decrease the content of TBIL in serum (P < 0.05). And the contents of GGT in serum of low dose rats were increased (P < 0.05). The bile flow was not changed by AEPM, but bile compositions of high dose male rats were obviously changed (TG increase, TBIL decrease, TBA decrease). The weights of liver and ratio of liver of the high dose rats were increased but showed no statistical significance. Pathologic examination displayed that there were only small pieces of necrosis in livers of several rats, without any severe disease.</p><p><b>CONCLUSION</b>AEPM can obviously injure bile duct epithelial cells, intervene liver cell functions and change bile compositions in rats, thus it is proved to induce cholestasis without severe liver injury.</p>
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Animaux , Femelle , Mâle , Rats , Administration par voie orale , Bile , Chimie , Cholestase , Foie , Anatomopathologie , Extraits de plantes , Toxicité , Polygonum , Toxicité , Rat Sprague-DawleyRésumé
<p><b>OBJECTIVE</b>For studying the pharmacokinetic of Yanshu injections in Beagel dogs, a sensitive and reproducible LC-MS method for quantitative determination of matrine, oxymatrine, sophocarpine and oxysophocarpine in dog's plasma were developed and validated using monocrotaline as an internal standard after iv of Yanshu injections (Sophorae Flavescentis Radix and Heterosmilacis Japonicae Rhizoma).</p><p><b>METHOD</b>The separation of plasma samples was performed on a CN column by isocratic elution with methanol-10 mmol x L(-1) NH4Ac-0.02% HCOOH-H2O 90:10 as the mobile phase. The plasma concentration of four kinds of alkaloids were calculated in dog plasta by detection of healthy dogs given Yanshu injection fluid after in twelve hours of plasma samples, All data of concentration-time of four kinds of alkaloids were treated with pharmacokinetics program DAS 2. 0.</p><p><b>RESULT</b>MT, OMT, SP and OSP have a good linear relationship in 0.01-16.0, 0.02-60.0, 0.01-4.0, 0.02-16.0 mg x L(-1), respectively. The average recoveries were more than 90% and the RSD of precision and stability of the test were less than 6.4% iv 1.2 g x kg(-1) Yanshu injection, four kinds of alkaloids in rats meet the two-compartment open pharmacokinetic model, Cmax and the concentration of the original liquid in the proportion of the basic line, the AUC(0-infinity) of matrine and oxymatrine, sophocarpine and oxysophocarpine compared to the original both in the proportion of liquid increases, the MRT(0-infinity) and t(1/2z) of matrine and sophocarpine were less than oxymatrine and oxysophocarpine; four kinds of alkaloids apparent volume of distribution matrine > oxymatrine, sophocarpine > oxysophocarpine.</p><p><b>CONCLUSION</b>A method with high recovery rate and good stabilitywas established to determine the blood concentration of MT, OMT, SP, OSP in Yanshu injection and applied in its pharmacokinetics successfully.</p>
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Animaux , Chiens , Alcaloïdes , Sang , Pharmacocinétique , Chromatographie en phase liquide , Médicaments issus de plantes chinoises , Chimie , Injections , Spectrométrie de masseRésumé
In this study, we successfully expanded a full length gene encoding the monooxygenase eytochrome P450 2C9 gene from human liver of Chinese Han by RT-PCR. Our findings indicated that except G-->T mutation at the 190th nucleotide site, the other nucleotide sequences are completely consistent with CYP2C9 (NM017460) in GenBank. The SDS-PAGE and Western-Blot analysis showed that the CYP 2C9 gene was successfully expressed in the host cell E. coli BL21 (DE3). Our current study lays the foundation for the evaluation of pre-clinical drug metabolism and safety in the future.
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Humains , Aryl hydrocarbon hydroxylases , Génétique , Chine , Ethnologie , Clonage moléculaire , Cytochrome P-450 CYP2C9 , Escherichia coli , Génétique , Métabolisme , Vecteurs génétiques , Génétique , Mutation ponctuelle , Polymorphisme génétique , Protéines recombinantes , GénétiqueRésumé
Purpose A RP-HPLC method was used to determine genioside and breviscapine in plasma and to study its pharmacokinetics in rat, respectively.Methods Rat plasma samples were collected after a single dose of Zhideng injection and pharmacokinetic parameters of genioside and breviscapine were estimated,respectively.Results A good linear relationship was obtained between 0.2-40.0 μg/mL for breviscapine, and 0.5-200.0 μg/mL for genioside.The recoveries from plasma were larger than 85%,and RSDs of inter-day asaay and intra-day assay were below 10%. The pharmacokinetic results showed that genioside and breviscapine were rapidly eliminated from plasma after iv administration of three doses of Zhideng injection.The mean half-life was 72.6 min and 21.6 min,respectively.Conclusion The established HPLC method was suitable for the pharmacokinetic study of genioside and breviscapine.