Objective:To investigate the propensity of plumbagin to inhibit the three isoforms of human cytochrome P450(CYP),ie.,CYP1A2,CYP2C19,and CYP3A4 using human liver microsomes in ritro.Methods:Inhibitory effects of plumba...Objective:To investigate the propensity of plumbagin to inhibit the three isoforms of human cytochrome P450(CYP),ie.,CYP1A2,CYP2C19,and CYP3A4 using human liver microsomes in ritro.Methods:Inhibitory effects of plumbagin on the three human CYP isoformswere investigated using pooled human liver microsomes.Phenacetin O-deethylation,omeprazole hydroxylation and nifedipine oxidation were used as selective substrates for CYP1A2,CYP2C19 and CYP3A4 activities,respectively.Concentrations of paracetamol,5-hydroxyomeprazole,and oxidized nifedipine were determined in microsomal incubation mixture using high performance liquid chromatography.Results:Plumbagin showed significantinhibitory effects on all CYP isoforms.but with the most potent activity on CYP2C19-mediated omeprazole hydroxylation.The IC50(concentration that inhibits enzyme activity by 50%) values of plumbagin and nootkatone(selective inhibitor) for CYP2C19 were(0.78±0.01) and(27.31±0.66) μM,respectively.The inhibitory activities on CYP1 A2-mediated phenacetin O-deethylation and CYP3A4-mediated nifedipine oxidation were moderate.The IC_(50) values of plumbagin and-naphthoflavone(selective inhibitor) for CYP1A2 were(1.39±0.01) and(0.02±.0.36) μM,respectively.The corresponding IC_(50) values of plumbagin and ketoconazole(selective inhibitor) for CYP3A4 were(2.37+0.10) and(0.18±0.06) μM,respectively.Conclusions:Clinical relevance of the interference of human drug metabolizing enzymes should be aware of for further development scheme of plumbagin as antimalarial drug when used in combination with other antimalarial drugs which are metabolized by these CYP isoforms.展开更多
Objective:To preliminarily investigate the possible role of prostaglandin D_2(PGD_2) in malaria infections.Methods:Blood and urinary samples(n=120 each) were collected from Thai patients with Plasmodium falciparum(P.f...Objective:To preliminarily investigate the possible role of prostaglandin D_2(PGD_2) in malaria infections.Methods:Blood and urinary samples(n=120 each) were collected from Thai patients with Plasmodium falciparum(P.falciparum) with moderate(n=26) and high(n=4) parasitemia,patients with Plasmodium vivax(P.vivax)(n=30),patients with fever associated with other infections(n=30),and healthy subjects(n=30).PGD_2 concentrations in plasma and urinary samples of healthy subjects,patients with fever associated with other infections and patients with malaria were determined using Prostaglandin D2-MOX express EIA kit(Cayman Chemical,USA).Results:The possible association between PGD_2 and malaria infections is clearly demonstrated with PGD_2 concentration in urine.The urinary PGD_2 concentrations were relatively high(about 5-fold) in patients with P.falciparum with moderate parasitemia and P.vivax infections compared with other groups.Furthermore,the concentration in patients with P.falciparum with moderate parasitemia and P.vivax infection were significantly higher than that in healthy subjects and patients with fever associated with other infections.Conclusions:Urinary PGD_2 concentrations may offer a more dependable and useful tool for predicting malaria severity.Confirmation is this preliminary finding is required with a larger sample size.展开更多
Objective:To evaluate the cytotoxic,apoptotic,mutagenic and immunomodulatory activities of Kaempferia galanga Linn.(KG)extract and ethyl-p-methoxycinnamate(EPMC)in vitro.Methods:The present study investigated the cyto...Objective:To evaluate the cytotoxic,apoptotic,mutagenic and immunomodulatory activities of Kaempferia galanga Linn.(KG)extract and ethyl-p-methoxycinnamate(EPMC)in vitro.Methods:The present study investigated the cytotoxic[using the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphe nyl-2-H-tetrazolium bromide test],apoptotic(using a mitochondrial membrane potential assay),mutagenic(using a micronucleus test)and immunomodulatory(using flow cytometry)activities of the ethanolic extract of KG and its bioactive component,EPMC,against two cholangiocarcinoma(CCA)cell lines,CL-6 and HuCCT1,and one normal human cell line,OUMS-36 T-1 F.Results:Both KG extract and EPMC exhibited moderate cytotoxic activity against both CCA cells.The cytotoxic activity was supported by their concentration-dependent induction of apoptosis.CL-6 was most sensitive(3–4 fold)and selective to 5-fluorouracil(5-FU),compared with KG extract and EPMC[median half inhibiting concentration(IC50)and selectivity index(SI)were 23.01 lg/mL and 17.32;78.41 lg/mL and 4.44;100.76 lg/mL and 2.20,respectively for 5-FU vs.KG extract vs.EPMC].HuCCT1 was relatively more sensitive and selective to 5-FU and EPMC than KG extract[median IC50 and SI were 66.03 lg/mL and6.04;60.90 lg/mL and 3.65;156.60 lg/mL and 2.23,respectively for 5-FU vs.EPMC vs.KG extract].EPMC produced relatively potent cytotoxic activity against polymorphonuclear cells(IC50=92.20 lg/mL).KG extract and EPMC exhibited concentration-dependent mutagenic activity,as well as inhibition of tumor necrosis factor-a and interleukin-6.Conclusion:Considering cytotoxic,apoptotic,immunomodulatory and mutagenic activities,further development of KG as a drug candidate is likely to focus on the oral pharmaceutical formulation of a standardized KG extract rather than isolated compounds.展开更多
Objective: To investigate the cytotoxic, apoptotic and inhibitory activities on cell migration and invasion of plumbagin in the human cholangiocarcinoma(CCA) cell line(CL-6) in comparison with human embryonic fibrobla...Objective: To investigate the cytotoxic, apoptotic and inhibitory activities on cell migration and invasion of plumbagin in the human cholangiocarcinoma(CCA) cell line(CL-6) in comparison with human embryonic fibroblast cell line(OUMS). Methods: Cytotoxicity activity was evaluated using MTT assay. Inhibitory effect on cell migration and invasion were investigated using label-free real-time cell analysis and QCM ECMatrix cell invasion chamber, respectively. Apoptotic activity was evaluated using flow cytometry and Cell Event? Caspase 3/7 assay. Results: Based on results of the cytotoxicity test in CL-6 cells, 50% inhibitory concentration(IC_(50), Mean±SD) values of plumbagin and the standard drug 5-fluorouracil were(24.00±3.33) and(1 036.00±137.77) μmol/L, respectively. The corresponding values for OUMS cells were(57.00±5.23) and(2 147.00±209.98) μmol/L, respectively. The selectivity index was 2.28. The inhibitory activities of plumbagin on cell migration and invasion were potent and concentration-dependent with IC_(50) of 25.0 μmol/L and complete inhibition at 25.0 μmol/L. Flow cytometry analysis showed that plumbagin at 12.5 μmol/L(half IC_(50)) induced CL-6 cell apoptosis(43.24% of control) through stimulation of caspase 3/7 activities. Complete cell apoptosis was observed at 12.5 μmol/L. Conclusions: The cytotoxic activity and inhibition of migration and invasion including apoptosis induction in the human CCA cell line(CL-6) suggest that plumbagin could be a promising candidate for CCA chemotherapeutics. However, its relatively low selective cytotoxic effect on CCA cells is a major concern.展开更多
Objective:To apply lectin affinity chromatography and glycoproteomics-based LC-MS/MS to preliminarily investigate the possible potential plasma biomarkers of Opisthorchis viverrini(OV)-associated CCA in OV/dimethylnit...Objective:To apply lectin affinity chromatography and glycoproteomics-based LC-MS/MS to preliminarily investigate the possible potential plasma biomarkers of Opisthorchis viverrini(OV)-associated CCA in OV/dimethylnitrosamine(DMN)-induced CCA hamster model.Methods:Nine Syrian hamsters were divided into 3 groups as follows(n=3 each):normal(healthy control group);OV group;and OV/DMN group(CCA group).Pooled plasma samples collected from animals in each group at the 6th month post-infection with OV metacercarae were subjected to glycoproteomics analysis.Glycoproteins in the pooled sample from each group were initially isolated by concanavilin A(Con A)-based affinity chromatography.The expression of glycoproteins isolated by both enrichment methods were determined using LCMS/MS.Results:Among the 24 Con A-binding glycoproteins isolated,two proteins,N-myc downstream regulated gene 1(NDRG1)and fetuin-B(FETUB)were found up-regulated only in the samples from the OV and control groups,but not in the OV/DMN(CCA)groups.On the other hand,one protein,i.e.,NSFL1 cofactor p47 isoform x3(NSFL1C)was found only in the samples from OV/DMN(CCA)and control groups,but not in the OV group.The remaining 21 proteins were upregulated in the samples from all groups.Conclusions:NDRG1,FETUB and NSFL1 C glycoproteins isolated by Con A-based affinity chromatography could be potential biomarkers for CCA.Plasma samples with negative for NDRG1 and FETUB proteins but positive for NSFL1 C are likely to be OV-associated CCA.Nevertheless,this conclusion remains to be confirmed whether this battery test can discriminate OV-associated CCA from other risk factors.展开更多
Objective: To investigate the role of human host heme-oxygenase-1(HO-1) in pathogenesis of cerebral malaria in the in vitro model,Methods: The effect of human host HO-1 [human brain microvascular endothelial cell(HBME...Objective: To investigate the role of human host heme-oxygenase-1(HO-1) in pathogenesis of cerebral malaria in the in vitro model,Methods: The effect of human host HO-1 [human brain microvascular endothelial cell(HBMEC)] on hemoglobin degradation in the co-culture model of HBMEC and ITG Plasmodium falciparum-infected red cells(i RBC) through measurement of the enzymatic products iron and bilirubin,Results: Following exposure to the HO-1 inducer Co PPIX at all concentrations,the HBMEC cells apoptosis occurred,which could be prominently observed at 15 μM of 3 h exposure,In contrast,there was no significant change in the morphology in the non-exposed i RBC at all concentrations and exposure time,This observation was in agreement with the levels of the enzymatic degradation products iron and bilirubin,of which the highest levels(106.03 and 1 753.54% of baseline level,respectively) were observed at 15 μM vs,20 μM at 3 h vs,24 h exposure,For the effect of the HO-1 inhibitor Zn PPIX,HBMEC cell morphology was mostly unchanged,but significant inhibitory effect on cell apoptosis was seen at 10 μM for the exposure period of 3 h(37.17% of baseline level),The degree of the inhibitory effect as reflected by the level of iron produced was not clearly observed(highest effect at 10 μM and 3 h exposure),Conclusions: Results provide at least in part,insight into the contribution of HO-1 on CM pathogenesis and need to be confirmed in animal model.展开更多
Objective: To preliminarily investigate the prevalence of HIV co-infection in patients with malaria in Mae Sot District, Tak Province of Thailand.Methods: The study was a retrospective study on blood samples collected...Objective: To preliminarily investigate the prevalence of HIV co-infection in patients with malaria in Mae Sot District, Tak Province of Thailand.Methods: The study was a retrospective study on blood samples collected from a total of 256 patients with malaria(all species and severity) who attended Mae Tao clinic for migrant workers, Tak Province during 2005-2007(148 samples) and 2010-2012(108 samples). Malaria diagnosis was performed based on microscopic examination of patients' blood smears. Chemiluminescent microparticle immunoassay and gel particle passive agglutination were employed for the detection of HIV antigen in patients' plasma. Results: Plasmodium falciparum(P. falciparum) and Plasmodium vivax(P. vivax) are the two predominant malaria species with the ratio of about 1: 1 to 1.5:1. Most of the P. falciparum cases were presented with acute uncomplicated signs and symptoms with highest parasitemia of 1 045 000 asexual parasites/μL bloods. The prevalence of malaria and HIV co-infection during 2005-2007 was 1.35%(2/148 cases, 1 each for P. falciparum and P. vivax co-infection), but was increased to 2.78%(3/108 cases, 2 and 1 for P. falciparum and P. vivax co-infection, respectively) during 2010-2012.Conclusions: The increasing trend of prevalence of malaria and HIV co-infection in Mae Sot, Tak province was of a great concern on either pharmacodynamics or pharmacokinetics aspect. The study in a larger numbers of malaria patients in different endemic areas throughout the country with different time periods is underway.展开更多
OBJECTIVE To formulate atractylodin-loaded poly(lactic-co-glycolic acid)(PLGA)nanoparticles and characterize the prepared nanoparticle formulation.METHODS The nanoparticle formulation was developed using solvent displ...OBJECTIVE To formulate atractylodin-loaded poly(lactic-co-glycolic acid)(PLGA)nanoparticles and characterize the prepared nanoparticle formulation.METHODS The nanoparticle formulation was developed using solvent displacement method.The encapsulation and loading efficiency were characterized and particle size,and zeta potential were determined by dynamic light scattering technique.Drug release was assessed in vitro.RESULTS The size(mean±SD of diameter) of the prepared atractylodin-loaded PLGA nanoparticles were(161.27 ± 1.87)nm with narrow size distribution(mean PDI:0.068±0.015) and zeta potential(28.83±0.35)mV.The encapsulation and loading efficiency were(48.31±0.83)% and(2.15±0.04)%,respectively.Drug release from atractylodin-loaded PLGA nanoparticles was observed up to(87.70±0.47)% in 72 h with biphasic manner.Moreover,the nanoparticles were found to be freely dispersible in water without aggregation.CONCLUSION Results suggest that PLGA nanoparticles may be used as an effective drug delivery system for atractylodin.The anti-cholangiocarcinoma activity of this nanoparticle formulation is required.展开更多
OBJECTIVE To identify potential cell signaling pathways and protein targets of the active compound isolated from Atracylodes lancea "atractylodin" in cholangiocarcinoma,using proteomics approach.METHODS The ...OBJECTIVE To identify potential cell signaling pathways and protein targets of the active compound isolated from Atracylodes lancea "atractylodin" in cholangiocarcinoma,using proteomics approach.METHODS The holangiocar-cinoma cell line was exposed with atractylodin for 3 and 6 h and the proteins from both intra-and extra-cellular components were extracted.The extract proteins were separated by SDS-PAGE and digested with trypsin.The LC-MS/MS was applied to identify proteins.Signaling pathways and protein expression were analyzed by MASCOT and STITCH software.RESULTS A total of 4,323 and 4,318 proteins were identified from intra-and extracellular components,respectively.Six intracellular proteins were linked with the signaling pathways(apoptosis,cell cycle control,and PI3K-AKT).Four extracellular proteins were linked with the signaling pathways(NF-κB and PI3K-AKT).CONCLUSION All these proteins will further study to confirm the link to the anticholangiocarcinoma ac.tivity of actractylodin.展开更多
The rhizome of Atractylodes Iancen(A.lancea)(Thunb.) DC.(AL)is extensively used in Chinese,Thai,and Japanese traditional medicines as crude extracts/decoctions or a component in various herbal formulations.Various pha...The rhizome of Atractylodes Iancen(A.lancea)(Thunb.) DC.(AL)is extensively used in Chinese,Thai,and Japanese traditional medicines as crude extracts/decoctions or a component in various herbal formulations.Various pharmacological activities of Al.and its major constituents have been demonstrated in ritro.ex ciro.and in animal models.Results from the toxicity studies in animal models suggest safety profile of AL,and its active constituents.Despite extensive use with positive impression in many diseases,there has not been a clinical study that can conclusively support its efficacy and safely profile in human.This review comprehensively summarizes current information on the pharmacological activities of AL and their active constituents including anticancer,anti-inflammatory,antimicrobial and antipyretic activities,as well as activities on central nervous,cardiovascular,and gastrointestinal systems.展开更多
Cholangiocarcinoma(CCA), the adenocarcinoma of the biliary duct, is commonly reported in Asia with the highest incidence in northeastern Thailand. Chemotherapy of this type of cancer is limited due to the lack of effe...Cholangiocarcinoma(CCA), the adenocarcinoma of the biliary duct, is commonly reported in Asia with the highest incidence in northeastern Thailand. Chemotherapy of this type of cancer is limited due to the lack of effective chemotherapeutic drugs. A series of previous studies support further research and development of Atractylodes lancea(Thunb) DC.(AL) as a potential candidate for the treatment of CCA as a crude ethanolic extract. In the present study, we aimed to develop an oral pharmaceutical formulation(capsule) of the standardized AL crude ethanolic extract for further clinical development in patients with CCA. Major steps included macroscopic and microscopic authentication of the AL rhizomes, preparation of standardized AL extract, preparation of oral pharmaceutical formulation(capsule) of the standardized AL extract, quantitative and qualitative analysis of the marker compound(atractylodin) in the formulated AL extract, evaluation of contaminations of heavy metals, pesticides residues, and microorganisms in the ground AL rhizomes and the formulated(capsule) powder of AL, physicochemical and pharmaceutical properties of the formulated AL extract/capsule, and cytotoxicity evaluation of the formulated AL extract. Results of all evaluations confirmed satisfactory pharmaceutical properties of oral(capsule) formulation of the standardized AL extract.展开更多
基金the financial support provided by Thammasat University Research Fund under the TU Research Scholar,Contract No 78/2557Commission on Higher Education,Ministry of Education of Thailand,Office of Higher Education Commission,Thammasat University(Excellence Center in Pharmacology and Molecular Biology of Malaria and Cholangiocarcinoma),Thammasat University and the Thailand Research Fund through a Royal Golden Jubilee Ph.D.scholarship to Wiriyaporn Sumsakul(Grant no.PHD/0326/2551)
文摘Objective:To investigate the propensity of plumbagin to inhibit the three isoforms of human cytochrome P450(CYP),ie.,CYP1A2,CYP2C19,and CYP3A4 using human liver microsomes in ritro.Methods:Inhibitory effects of plumbagin on the three human CYP isoformswere investigated using pooled human liver microsomes.Phenacetin O-deethylation,omeprazole hydroxylation and nifedipine oxidation were used as selective substrates for CYP1A2,CYP2C19 and CYP3A4 activities,respectively.Concentrations of paracetamol,5-hydroxyomeprazole,and oxidized nifedipine were determined in microsomal incubation mixture using high performance liquid chromatography.Results:Plumbagin showed significantinhibitory effects on all CYP isoforms.but with the most potent activity on CYP2C19-mediated omeprazole hydroxylation.The IC50(concentration that inhibits enzyme activity by 50%) values of plumbagin and nootkatone(selective inhibitor) for CYP2C19 were(0.78±0.01) and(27.31±0.66) μM,respectively.The inhibitory activities on CYP1 A2-mediated phenacetin O-deethylation and CYP3A4-mediated nifedipine oxidation were moderate.The IC_(50) values of plumbagin and-naphthoflavone(selective inhibitor) for CYP1A2 were(1.39±0.01) and(0.02±.0.36) μM,respectively.The corresponding IC_(50) values of plumbagin and ketoconazole(selective inhibitor) for CYP3A4 were(2.37+0.10) and(0.18±0.06) μM,respectively.Conclusions:Clinical relevance of the interference of human drug metabolizing enzymes should be aware of for further development scheme of plumbagin as antimalarial drug when used in combination with other antimalarial drugs which are metabolized by these CYP isoforms.
基金funded by Thailand Research Fund-Thammasat University Joint Fund and Graduated Student Grant to P.Thongdee(No.PHD/0365/2552)
文摘Objective:To preliminarily investigate the possible role of prostaglandin D_2(PGD_2) in malaria infections.Methods:Blood and urinary samples(n=120 each) were collected from Thai patients with Plasmodium falciparum(P.falciparum) with moderate(n=26) and high(n=4) parasitemia,patients with Plasmodium vivax(P.vivax)(n=30),patients with fever associated with other infections(n=30),and healthy subjects(n=30).PGD_2 concentrations in plasma and urinary samples of healthy subjects,patients with fever associated with other infections and patients with malaria were determined using Prostaglandin D2-MOX express EIA kit(Cayman Chemical,USA).Results:The possible association between PGD_2 and malaria infections is clearly demonstrated with PGD_2 concentration in urine.The urinary PGD_2 concentrations were relatively high(about 5-fold) in patients with P.falciparum with moderate parasitemia and P.vivax infections compared with other groups.Furthermore,the concentration in patients with P.falciparum with moderate parasitemia and P.vivax infection were significantly higher than that in healthy subjects and patients with fever associated with other infections.Conclusions:Urinary PGD_2 concentrations may offer a more dependable and useful tool for predicting malaria severity.Confirmation is this preliminary finding is required with a larger sample size.
基金funded by Center of Excellence in Pharmacology and Molecular Biology of Malaria and Cholangiocarcinoma of Thammasat UniversityThammasat University Research fund under the TU Research Scholar(Contract No.03/2561)。
文摘Objective:To evaluate the cytotoxic,apoptotic,mutagenic and immunomodulatory activities of Kaempferia galanga Linn.(KG)extract and ethyl-p-methoxycinnamate(EPMC)in vitro.Methods:The present study investigated the cytotoxic[using the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphe nyl-2-H-tetrazolium bromide test],apoptotic(using a mitochondrial membrane potential assay),mutagenic(using a micronucleus test)and immunomodulatory(using flow cytometry)activities of the ethanolic extract of KG and its bioactive component,EPMC,against two cholangiocarcinoma(CCA)cell lines,CL-6 and HuCCT1,and one normal human cell line,OUMS-36 T-1 F.Results:Both KG extract and EPMC exhibited moderate cytotoxic activity against both CCA cells.The cytotoxic activity was supported by their concentration-dependent induction of apoptosis.CL-6 was most sensitive(3–4 fold)and selective to 5-fluorouracil(5-FU),compared with KG extract and EPMC[median half inhibiting concentration(IC50)and selectivity index(SI)were 23.01 lg/mL and 17.32;78.41 lg/mL and 4.44;100.76 lg/mL and 2.20,respectively for 5-FU vs.KG extract vs.EPMC].HuCCT1 was relatively more sensitive and selective to 5-FU and EPMC than KG extract[median IC50 and SI were 66.03 lg/mL and6.04;60.90 lg/mL and 3.65;156.60 lg/mL and 2.23,respectively for 5-FU vs.EPMC vs.KG extract].EPMC produced relatively potent cytotoxic activity against polymorphonuclear cells(IC50=92.20 lg/mL).KG extract and EPMC exhibited concentration-dependent mutagenic activity,as well as inhibition of tumor necrosis factor-a and interleukin-6.Conclusion:Considering cytotoxic,apoptotic,immunomodulatory and mutagenic activities,further development of KG as a drug candidate is likely to focus on the oral pharmaceutical formulation of a standardized KG extract rather than isolated compounds.
基金supported by Thammasat University research grant(Grant No.20/2556:Ms.Luxana Panrit)Thammasat University Center of Excellence in Pharmacology and Molecular Biology of Malaria and Cholangiocarcinoma+1 种基金The Commission of Higher Education,Ministry of Education of Thailand(National University Project)National Research Council of Thailand
文摘Objective: To investigate the cytotoxic, apoptotic and inhibitory activities on cell migration and invasion of plumbagin in the human cholangiocarcinoma(CCA) cell line(CL-6) in comparison with human embryonic fibroblast cell line(OUMS). Methods: Cytotoxicity activity was evaluated using MTT assay. Inhibitory effect on cell migration and invasion were investigated using label-free real-time cell analysis and QCM ECMatrix cell invasion chamber, respectively. Apoptotic activity was evaluated using flow cytometry and Cell Event? Caspase 3/7 assay. Results: Based on results of the cytotoxicity test in CL-6 cells, 50% inhibitory concentration(IC_(50), Mean±SD) values of plumbagin and the standard drug 5-fluorouracil were(24.00±3.33) and(1 036.00±137.77) μmol/L, respectively. The corresponding values for OUMS cells were(57.00±5.23) and(2 147.00±209.98) μmol/L, respectively. The selectivity index was 2.28. The inhibitory activities of plumbagin on cell migration and invasion were potent and concentration-dependent with IC_(50) of 25.0 μmol/L and complete inhibition at 25.0 μmol/L. Flow cytometry analysis showed that plumbagin at 12.5 μmol/L(half IC_(50)) induced CL-6 cell apoptosis(43.24% of control) through stimulation of caspase 3/7 activities. Complete cell apoptosis was observed at 12.5 μmol/L. Conclusions: The cytotoxic activity and inhibition of migration and invasion including apoptosis induction in the human CCA cell line(CL-6) suggest that plumbagin could be a promising candidate for CCA chemotherapeutics. However, its relatively low selective cytotoxic effect on CCA cells is a major concern.
基金supported by the National Research University Project(NRU)of Thailand Office of Higher Education Commission,Ministry of Education of Thailand and Thammasat University(Excellence Center in Pharmacology and Molecular Biology of Malaria and Cholangiocarcinoma)
文摘Objective:To apply lectin affinity chromatography and glycoproteomics-based LC-MS/MS to preliminarily investigate the possible potential plasma biomarkers of Opisthorchis viverrini(OV)-associated CCA in OV/dimethylnitrosamine(DMN)-induced CCA hamster model.Methods:Nine Syrian hamsters were divided into 3 groups as follows(n=3 each):normal(healthy control group);OV group;and OV/DMN group(CCA group).Pooled plasma samples collected from animals in each group at the 6th month post-infection with OV metacercarae were subjected to glycoproteomics analysis.Glycoproteins in the pooled sample from each group were initially isolated by concanavilin A(Con A)-based affinity chromatography.The expression of glycoproteins isolated by both enrichment methods were determined using LCMS/MS.Results:Among the 24 Con A-binding glycoproteins isolated,two proteins,N-myc downstream regulated gene 1(NDRG1)and fetuin-B(FETUB)were found up-regulated only in the samples from the OV and control groups,but not in the OV/DMN(CCA)groups.On the other hand,one protein,i.e.,NSFL1 cofactor p47 isoform x3(NSFL1C)was found only in the samples from OV/DMN(CCA)and control groups,but not in the OV group.The remaining 21 proteins were upregulated in the samples from all groups.Conclusions:NDRG1,FETUB and NSFL1 C glycoproteins isolated by Con A-based affinity chromatography could be potential biomarkers for CCA.Plasma samples with negative for NDRG1 and FETUB proteins but positive for NSFL1 C are likely to be OV-associated CCA.Nevertheless,this conclusion remains to be confirmed whether this battery test can discriminate OV-associated CCA from other risk factors.
基金supported by The Commission on Higher Education,Ministry of Education of Thailand,The National Research University Project of Thailand(NRU)Office of Higher Education Commission,Thammasat University(Center of Excellence in Pharmacology and Molecular Biology of Malaria and Cholangiocarcinoma)+1 种基金Liverpool School of Tropical Medicine,University of Liverpool,UKThe Royal Golden Jubilee PhD Programme,Thailand Research Fund-Thammasat University Joint Fund and Graduated Student Grant to P.Thongdee(Grant No.PHD/0365/2552)
文摘Objective: To investigate the role of human host heme-oxygenase-1(HO-1) in pathogenesis of cerebral malaria in the in vitro model,Methods: The effect of human host HO-1 [human brain microvascular endothelial cell(HBMEC)] on hemoglobin degradation in the co-culture model of HBMEC and ITG Plasmodium falciparum-infected red cells(i RBC) through measurement of the enzymatic products iron and bilirubin,Results: Following exposure to the HO-1 inducer Co PPIX at all concentrations,the HBMEC cells apoptosis occurred,which could be prominently observed at 15 μM of 3 h exposure,In contrast,there was no significant change in the morphology in the non-exposed i RBC at all concentrations and exposure time,This observation was in agreement with the levels of the enzymatic degradation products iron and bilirubin,of which the highest levels(106.03 and 1 753.54% of baseline level,respectively) were observed at 15 μM vs,20 μM at 3 h vs,24 h exposure,For the effect of the HO-1 inhibitor Zn PPIX,HBMEC cell morphology was mostly unchanged,but significant inhibitory effect on cell apoptosis was seen at 10 μM for the exposure period of 3 h(37.17% of baseline level),The degree of the inhibitory effect as reflected by the level of iron produced was not clearly observed(highest effect at 10 μM and 3 h exposure),Conclusions: Results provide at least in part,insight into the contribution of HO-1 on CM pathogenesis and need to be confirmed in animal model.
基金Support by Thammasat University(Center of Excellence in Pharmacology and Molecular Biology of Malaria and Cholangiocarcinoma),the National Research Council of Thailand(Grant No.034/2556)the Higher Education Commission of Thailand(Grant No.036/554)
文摘Objective: To preliminarily investigate the prevalence of HIV co-infection in patients with malaria in Mae Sot District, Tak Province of Thailand.Methods: The study was a retrospective study on blood samples collected from a total of 256 patients with malaria(all species and severity) who attended Mae Tao clinic for migrant workers, Tak Province during 2005-2007(148 samples) and 2010-2012(108 samples). Malaria diagnosis was performed based on microscopic examination of patients' blood smears. Chemiluminescent microparticle immunoassay and gel particle passive agglutination were employed for the detection of HIV antigen in patients' plasma. Results: Plasmodium falciparum(P. falciparum) and Plasmodium vivax(P. vivax) are the two predominant malaria species with the ratio of about 1: 1 to 1.5:1. Most of the P. falciparum cases were presented with acute uncomplicated signs and symptoms with highest parasitemia of 1 045 000 asexual parasites/μL bloods. The prevalence of malaria and HIV co-infection during 2005-2007 was 1.35%(2/148 cases, 1 each for P. falciparum and P. vivax co-infection), but was increased to 2.78%(3/108 cases, 2 and 1 for P. falciparum and P. vivax co-infection, respectively) during 2010-2012.Conclusions: The increasing trend of prevalence of malaria and HIV co-infection in Mae Sot, Tak province was of a great concern on either pharmacodynamics or pharmacokinetics aspect. The study in a larger numbers of malaria patients in different endemic areas throughout the country with different time periods is underway.
基金supported by Center of Excellence in Pharmacology and Molecular Biology of Malaria and Cholangiocarcinoma,Chulabhorn International College of Medicine,Thammasat University the National Research University Project of Thailand(NRU)
文摘OBJECTIVE To formulate atractylodin-loaded poly(lactic-co-glycolic acid)(PLGA)nanoparticles and characterize the prepared nanoparticle formulation.METHODS The nanoparticle formulation was developed using solvent displacement method.The encapsulation and loading efficiency were characterized and particle size,and zeta potential were determined by dynamic light scattering technique.Drug release was assessed in vitro.RESULTS The size(mean±SD of diameter) of the prepared atractylodin-loaded PLGA nanoparticles were(161.27 ± 1.87)nm with narrow size distribution(mean PDI:0.068±0.015) and zeta potential(28.83±0.35)mV.The encapsulation and loading efficiency were(48.31±0.83)% and(2.15±0.04)%,respectively.Drug release from atractylodin-loaded PLGA nanoparticles was observed up to(87.70±0.47)% in 72 h with biphasic manner.Moreover,the nanoparticles were found to be freely dispersible in water without aggregation.CONCLUSION Results suggest that PLGA nanoparticles may be used as an effective drug delivery system for atractylodin.The anti-cholangiocarcinoma activity of this nanoparticle formulation is required.
基金supported by Center of Excellence in Pharmacology and Molecular Biology of Malaria and Cholangiocarcinoma,Chulabhorn International College of Medicine,Thammasat University The National Research University Project of Thailand(NRU)
文摘OBJECTIVE To identify potential cell signaling pathways and protein targets of the active compound isolated from Atracylodes lancea "atractylodin" in cholangiocarcinoma,using proteomics approach.METHODS The holangiocar-cinoma cell line was exposed with atractylodin for 3 and 6 h and the proteins from both intra-and extra-cellular components were extracted.The extract proteins were separated by SDS-PAGE and digested with trypsin.The LC-MS/MS was applied to identify proteins.Signaling pathways and protein expression were analyzed by MASCOT and STITCH software.RESULTS A total of 4,323 and 4,318 proteins were identified from intra-and extracellular components,respectively.Six intracellular proteins were linked with the signaling pathways(apoptosis,cell cycle control,and PI3K-AKT).Four extracellular proteins were linked with the signaling pathways(NF-κB and PI3K-AKT).CONCLUSION All these proteins will further study to confirm the link to the anticholangiocarcinoma ac.tivity of actractylodin.
文摘The rhizome of Atractylodes Iancen(A.lancea)(Thunb.) DC.(AL)is extensively used in Chinese,Thai,and Japanese traditional medicines as crude extracts/decoctions or a component in various herbal formulations.Various pharmacological activities of Al.and its major constituents have been demonstrated in ritro.ex ciro.and in animal models.Results from the toxicity studies in animal models suggest safety profile of AL,and its active constituents.Despite extensive use with positive impression in many diseases,there has not been a clinical study that can conclusively support its efficacy and safely profile in human.This review comprehensively summarizes current information on the pharmacological activities of AL and their active constituents including anticancer,anti-inflammatory,antimicrobial and antipyretic activities,as well as activities on central nervous,cardiovascular,and gastrointestinal systems.
基金Financial support by National Research Council of Thailand(NRCT)Center of Excellence in Pharmacology and Molecular Biology of Malaria+1 种基金Cholangiocarcinoma and Chulabhorn International College of Medicine(CICM)Thammasat University,Thailand
文摘Cholangiocarcinoma(CCA), the adenocarcinoma of the biliary duct, is commonly reported in Asia with the highest incidence in northeastern Thailand. Chemotherapy of this type of cancer is limited due to the lack of effective chemotherapeutic drugs. A series of previous studies support further research and development of Atractylodes lancea(Thunb) DC.(AL) as a potential candidate for the treatment of CCA as a crude ethanolic extract. In the present study, we aimed to develop an oral pharmaceutical formulation(capsule) of the standardized AL crude ethanolic extract for further clinical development in patients with CCA. Major steps included macroscopic and microscopic authentication of the AL rhizomes, preparation of standardized AL extract, preparation of oral pharmaceutical formulation(capsule) of the standardized AL extract, quantitative and qualitative analysis of the marker compound(atractylodin) in the formulated AL extract, evaluation of contaminations of heavy metals, pesticides residues, and microorganisms in the ground AL rhizomes and the formulated(capsule) powder of AL, physicochemical and pharmaceutical properties of the formulated AL extract/capsule, and cytotoxicity evaluation of the formulated AL extract. Results of all evaluations confirmed satisfactory pharmaceutical properties of oral(capsule) formulation of the standardized AL extract.
