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1.
Zhongguo Zhong Yao Za Zhi ; 46(1): 196-205, 2021 Jan.
Artigo em Chinês | MEDLINE | ID: mdl-33645071

RESUMO

The present study is to investigate the absorption characteristics of the main components in Polygonum orientale extract in normal and isoproterenol-induced myocardial ischemia model rats with everted intestinal sac models. Intestinal sac fluid samples were collected in different part of intestine(duodenum, jejunum, ileum, colon) at different time after administration of different concentration of P. orientale extract(5.0,10.0, 20.0 mg·mL~(-1)). An UPLC-TQD method was employed for the determination of six components including orientin, isoorientin, vitexin, protocatechuic acid, kaempferol-3-O-ß-D-glucoside and quercitrin in the intestinal sac samples. The absorption rate and cumulative absorption were calculated to analyze the intestinal absorption characteristics of six components in normal and myocardial ischemia model rats. The P-glycoprotein(P-gp) inhibitor was applied to investigate influence of intestinal absorption of six components in P. orientale extract. The results showed that the main absorption sites were concentrated on the duodenum at low concentration, while they were the colon at the medium concentration and the ileum at high concentration in control groups. In the condition of myocardial ischemia model, the main absorption sites focus on the ileum and jejunum at low concentration; the main absorption sites were in the ileum at the medium concentration and main absorption sites were the duodenum and ileum at high concentration. Compared with the normal group, the absorption rate and cumulative absorption of the six components significantly decreased in the model group. P-gp inhibitor markedly increased the absorption rate and cumulative absorption of six components in the model group, inferring that the 6 components may be the substrates of P-gp, and the mechanism needs further study. In this study, it is revealed that the six components of P. orientale extract can be absorbed into the intestinal sac, and it is an effective method to assess the intestinal absorption characteristics of P. orientale extract through everted intestinal sac model, providing data support for the clinical application and further development of P. orientale.


Assuntos
Isquemia Miocárdica , Polygonum , Animais , Absorção Intestinal , Intestinos , Isoproterenol , Isquemia Miocárdica/induzido quimicamente , Ratos , Ratos Sprague-Dawley
2.
J Tradit Chin Med ; 41(1): 51-58, 2021 02.
Artigo em Inglês | MEDLINE | ID: mdl-33522197

RESUMO

OBJECTIVE: To investigate the efficacy of water fraction from Dioscorea cirrhosa (WF) on oxidative damage and apoptosis of cardiomyocytes induced by H2O2, and to study its mechanism. METHODS: Cell viability was measured by the MST assay kit. The content of malondialdehyde (MDA), release of lactate dehydrogenase (LDH) and activity of catalase (CAT) and superoxide dismutase (SOD) were detected by biochemical kit. The content of reactive oxygen species (ROS) was assessed by nonfluorescent probe 2' ,7'-dichlorofluorescin diacetate (DCFH-DA). JC-1 was used to analyze the mitochondrial membrane potential (mtΔΨ) and Annexin-V-FITC/PI staining was applied to assess apoptosis of H9c2 by flow cytometry. Moreover, the expression of B-cell lymphoma-2 (Bcl-2), Bcl-2-associated X (Bax), caspase-3, caspase-9, cleaved-caspase-3 and cleaved-caspase -9 proteins was determined by western blot analysis. RESULTS: WF increased cell viability and decreased LDH leakage in H9c2 cells exposed to H2O2. WF treatment decreased ROS and MDA level, enhanced SOD and CAT activities, improved mtΔΨ and inhibited apoptosis. Western blot analysis demonstrated that the ratio of Bcl-2/Bax was increased and the expression cleaved-caspase-3, caspase-3, cleaved-caspase-9 and caspase-9 were decreased in group treated with WF. CONCLUSION: WF protects H9c2 myocardial cells on H2O2-induced oxidative stress and apoptosis by scavenging ROS, improving antioxidant capacity, protecting mitochondrial and regulating the proteins expression related to apoptosis.

3.
Genes (Basel) ; 12(2)2021 Feb 09.
Artigo em Inglês | MEDLINE | ID: mdl-33572241

RESUMO

An emerging poplar canker caused by the gram-negative bacterium, Lonsdalea populi, has led to high mortality of hybrid poplars Populus × euramericana in China and Europe. The molecular bases of pathogenicity and bark adaptation of L. populi have become a focus of recent research. This study revealed the whole genome sequence and identified putative virulence factors of L. populi. A high-quality L. populi genome sequence was assembled de novo, with a genome size of 3,859,707 bp, containing approximately 3434 genes and 107 RNAs (75 tRNA, 22 rRNA, and 10 ncRNA). The L. populi genome contained 380 virulence-associated genes, mainly encoding for adhesion, extracellular enzymes, secretory systems, and two-component transduction systems. The genome had 110 carbohydrate-active enzyme (CAZy)-coding genes and putative secreted proteins. The antibiotic-resistance database annotation listed that L. populi was resistant to penicillin, fluoroquinolone, and kasugamycin. Analysis of comparative genomics found that L. populi exhibited the highest homology with the L. britannica genome and L. populi encompassed 1905 specific genes, 1769 dispensable genes, and 1381 conserved genes, suggesting high evolutionary diversity and genomic plasticity. Moreover, the pan genome analysis revealed that the N-5-1 genome is an open genome. These findings provide important resources for understanding the molecular basis of the pathogenicity and biology of L. populi and the poplar-bacterium interaction.

4.
Artigo em Inglês | MEDLINE | ID: mdl-33062020

RESUMO

Background: Gukang capsule (GKC) is a traditional Chinese medicine formulation which has been used extensively in the clinical treatment of bone fractures. However, the mechanisms underlying its effects on fracture healing remain unclear. Methods: In this study we used a rabbit radius fracture model, and we measured the serum content of bone alkaline phosphatase (ALP), calcium, and phosphorus and examined pathology of the fracture site as indicators of the fracture healing effects of GKC. SaOS-2 human osteosarcoma cells were used to measure (i) ALP activity, (ii) ornithine transcarbamylase (OTC), calcium, and mineralization levels, (iii) the expression of osteogenic-related genes, that is, runt-related transcription factor 2 (RUNX2), bone morphogenetic protein 2 (BMP2), collagen I (COL-I), osteopontin (OPN), OTC, and osterix (Osx), and (iv) the expression of key proteins in the Wnt/ß-catenin and BMP/SMAD signaling pathways to study the mechanisms by which GKC promotes fracture healing. Results: We found that GKC effectively promotes radius fracture healing in rabbits and enhances ALP activity, increases OTC and calcium levels, and stimulates the formation of mineralized nodules in SaOS-2 cells. Moreover, COL-I, OTC, Osx, BMP2, and OPN expression levels were higher in SaOS-2 cells treated with GKC than control cells. GKC upregulates glycogen synthase kinase 3ß (GSK3ß) phosphorylation and Smad1/5 and ß-catenin protein levels, thereby activating Wnt/ß-catenin and BMP/Smad signaling pathways. Inhibitors of the Wnt/ß-catenin and BMP/Smad signaling pathways (DKK1 and Noggin, respectively) suppress the osteogenic effects of GKC. Conclusions: GKC promotes fracture healing by activating the Wnt/ß-catenin and BMP/Smad signaling pathways and increasing osteoprotegerin (OPG) secretion by osteoblasts (OBs), which prevents receptor activator of nuclear factor kappa B ligand (RANKL) binding to RANK.

5.
PeerJ ; 8: e10046, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33024649

RESUMO

Chlorophyll fluorescence (ChlF) has been used to understand photosynthesis and its response to climate change, particularly with satellite-based data. However, it remains unclear how the ChlF ratio and photosynthesis are linked at the leaf level under drought stress. Here, we examined the link between ChlF ratio and photosynthesis at the leaf level by measuring photosynthetic traits, such as net CO2 assimilation rate (An), the maximum carboxylation rate of Rubisco (Vcmax), the maximum rate of electron transport (Jmax), stomatal conductance (gs) and total chlorophyll content (Chlt). The ChlF ratio of the leaf level such as maximum quantum efficiency of PSII (Fv/Fm) is based on fluorescence kinetics. ChlF intensity ratio (LD685/LD740) based on spectrum analysis was obtained. We found that a combination of the stomatal limitation, non-stomatal limitation, and Chlt regulated leaf photosynthesis under drought stress, while Jmax and Chlt governed the ChlF ratio. A significant link between the ChlF ratio and An was found under drought stress while no significant correlation in the control, which indicated that drought stress strengthens the link between the ChlF ratio and photosynthetic traits. These results suggest that the ChlF ratio can be a powerful tool to track photosynthetic traits of terrestrial ecosystems under drought stress.

6.
Mol Plant Pathol ; 21(11): 1451-1466, 2020 11.
Artigo em Inglês | MEDLINE | ID: mdl-32954659

RESUMO

Reactive oxygen/nitrogen species (ROS/RNS) play a fundamental role in plant-fungal interactions. How pathogenic fungi manipulate plant-derived ROS/RNS is of importance to the outcomes of these interactions. In this study, we explored the individual and combined contributions of three transcription factors, VdAtf1, VdYap1, and VdSkn7, in the response to ROS/RNS, microsclerotia formation, and virulence in the plant wilt pathogen Verticillium dahliae. We showed that VdYap1 is essential for ROS response. Additionally, mutants lacking any combination of the three genes shared significant hypersensitivity to nitro-oxidative stress like sodium nitroprusside dehydrate and double deletions lacking VdYap1 and VdAtf1 resulted in further increased sensitivity to ROS. Double deletion of VdAtf1 and VdSkn7 reduced melanin production and virulence while simultaneous lack of VdSkn7 and VdYap1 disrupted nitrogen metabolism and ROS resistance. Finally, comparison of transcriptional profiles of the respective single or double mutants in response to nitro-oxidative stress revealed that the three transcription factors are involved in denitrification of nitrated alkanes and lipids to protect against nitro-oxidative stress. Taken together, our results demonstrate convergent and distinctive functions of VdYap1, VdAtf1, and VdSkn7 in V. dahliae, and provide new data on their roles in response to ROS/RNS in fungi.

7.
Biomed Pharmacother ; 130: 110537, 2020 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-32717630

RESUMO

Rhizoma Bletillae, the tubes of Bletilla striata, has been traditionally used in China as a hemostatic agent. In preliminary studies, the major active fraction responsible for its hemostatic effect have been confirmed to be Rhizoma Bletillae polysaccharide (RBp), but the hemostatic mechanism of action of RBp is still unknown.The main aim of this study was to clarify its mechanism of hemostatic effect. RBp was prepared by 80 % ethanol precipitation of the water extract of Rhizoma Bletillae followed by the Sevag method to remove proteins. The average molecular weight (Mw) of the crude RBp maintained at a range of 30.06-200 KDa. The hemostatic effects of RBp were evaluated by testing its effect on the platelet aggregation of rat platelet-rich plasma (PRP). PRP was dealt with different concentrations of RBp and platelet aggregation was measured by the turbidimetric method. The hemostatic mechanism of RBp was investigated by examining its effect on platelet shape, platelet secretion, and activation of related receptors (P2Y1, P2Y12 and TXA2) by electron microscopy and the turbidimetric method. RBp significantly enhanced the platelet aggregations at concentrations of 50-200 mg/L in a concentration-dependent manner. The inhibitory rate of platelet aggregation was significantly increased by apyrase and Ro31-8220 in a concentration-dependent manner, while RBp-induced platelet aggregation was completely inhibited by P2Y1, P2Y12 and the PKC receptor antagonists. However, the aggregation was not sensitive to TXA2. RBp, the active ingredients of Rhizoma Bletillae responsible for its hemostatic effect, could significantly accelerate the platelet aggregation and shape change. The hemostatic mechanism may involve activation of the P2Y1, P2Y12, and PKC receptors in the adenosine diphosphate (ADP) receptor signaling pathway.

8.
Zhongguo Zhong Yao Za Zhi ; 45(11): 2626-2633, 2020 Jun.
Artigo em Chinês | MEDLINE | ID: mdl-32627498

RESUMO

This study aims to reveal the pharmacokinetics of Shuganning Injection in normal rats. In this experiment,ultra-high performance liquid chromatography-electrospray-tandem mass spectrometry( UPLC-ESI-MS/MS) was used to establish an analytical method for simultaneous determination of chlorogenic acid,gardenioside,oroxylin A and baicalin in rat plasma. Then,the non-compartmental model( NCA) in Phoenix WinN onL in 6. 4 software was used to fit pharmacokinetic parameters. The methodological validation showed that the linear relationship of the components in rat plasma samples were good( r>0. 995). The recovery rate and matrix effect of plasma samples with low,middle and high concentration were 79. 14%-101. 4%. The intra-day and inter-day precision,accuracy and stability meet the requirements of biological sample analysis. The half-life( t1/2) of chlorogenic acid,gardenioside,oroxylin A did not change significantly and the area under blood concentration-time curve( AUC0-t) is proportional to the dose,which suggested that three components showed a linear kinetic characteristics,but baicalin showed nonlinear kinetic characteristics. Moreover,the retention time of each component in rats was short. The established UPLC-MS/MS quantitative analysis method is rapid,sensitive and accurate,which can be used for the determination of chlorogenic acid,gardenioside,oroxylin A and baicalin in rat plasma and pharmacokinetic study of Shuganning Injection.


Assuntos
Plasma , Espectrometria de Massas em Tandem , Animais , Ácido Clorogênico , Cromatografia Líquida de Alta Pressão , Cromatografia Líquida , Ratos , Ratos Sprague-Dawley , Reprodutibilidade dos Testes
9.
Photochem Photobiol Sci ; 19(7): 966-975, 2020 Jul 15.
Artigo em Inglês | MEDLINE | ID: mdl-32525187

RESUMO

One-dimensional TiO2@C nanocables with a heterophase junction have been successfully prepared by coating brookite@anatase TiO2 with a thin layer of hydrothermal carbon (HTC). Compared with anatase TiO2, the biphase brookite@anatase structure can reduce the recombination rate of the excited electron/hole pairs of TiO2. The HTC coating not only enhances the adsorption capability of the TiO2 catalyst for organic pollutants but also facilitates photogenerated electron transfer to further increase its photocatalytic activity. Therefore, compared with anatase TiO2, brookite@anatase TiO2, and TiO2@C, the brookite@anatase TiO2@C shows the highest photocatalytic activity for the photodegradation of tetracycline (TC) under the irradiation of UV-visible light. Moreover, ˙O2 has been proved to be the predominant active species for the photodegradation of TC, and the photocatalytic mechanism of brookite@anatase TiO2@C nanocables has also been proposed.

10.
Bioorg Chem ; 101: 104025, 2020 08.
Artigo em Inglês | MEDLINE | ID: mdl-32599368

RESUMO

Metastatic progression of cancer is a complex and clinically daunting process, with migration, invasion and angiogenesis being the key features. Tetrandrine (TET) is a typical dibenzylisoquinoline alkaloid with promising anti-tumor activity. In our previous work, a number of TET derivatives were designed and synthesized with obvious anti-proliferation activities against cancer cells, however, the anti-metastatic effects of these compounds were not evaluated. In the current investigation, five TET derivatives (8, 18, 32, 71, and 72) with pronounced anti-proliferative activities (IC50 values of 1.00, 1.91, 3.43, 3.78, and 1.93 µM, respectively) against human umbilical vein endothelial cells (HUVECs) were screened out. Scratch assays showed that these compounds significantly suppressed the migration of HUVECs and induced their apoptosis. Among them, derivatives 8 and 72 obviously inhibited the proliferation, colony formation and invasion of HCT-15 cells. Tube formation assays revealed that 4 µM of 8 or 72 remarkably inhibited the tube forming capacity of HUVECs. Moreover, 8 and 72 surpressed the formation of filopodia in HUVECs and severely impaired their motility. Both compounds effectively inhibited the angiogenesis in the zebrafish model with low toxicities in vivo. These results indicated that TET derivatives 8 and 72 are promising anti-metastatic inhibitors.

11.
Xenobiotica ; 50(10): 1251-1257, 2020 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-32375563

RESUMO

Hepatocellular carcinoma (HCC) is a malignancy of liver cells. Recent studies have shown that HCC patients often have changes in the activities of transporters and metabolic enzymes, which can considerably affect drug pharmacokinetics and lead to drug toxicity. Doxorubicin (DOX) has been frequently administered in chemotherapy for HCC, but to our knowledge, the effects of HCC on the pharmacokinetics of DOX are unknown.In the present study, following intravenous administration of DOX in diethylnitrosamine-induced HCC rats, the plasma concentration was determined by a UPLC/MS/MS method. The expression of metabolic enzyme and transporters (p-gp, cbr1 and slc22a16) was analyzed by qRT-PCR and western blot.The results showed that the pharmacokinetic parameters AUC, T1/2, K12 and K21 of DOX were markedly increased, the K10 and CL were significantly decreased in HCC rats. The expression of cbr1 and slc22a16 was markedly decreased, while p-gp was significantly upregulated in HCC rats.These findings suggest that HCC could significantly alter the pharmacokinetic profile of DOX, which may be associated with the decreased expression of cbr1 and slc22a16 rather than the upregulation of p-gp expression.


Assuntos
Doxorrubicina/farmacocinética , Animais , Carcinoma Hepatocelular , Dietilnitrosamina , Neoplasias Hepáticas , Ratos
12.
Nanotechnology ; 31(37): 375604, 2020 Sep 11.
Artigo em Inglês | MEDLINE | ID: mdl-32454467

RESUMO

Using citric acid (CA) and 1,5-naphthalenedisulfonic acid (NDSA) as the structure-directing agent, a hierarchical flower-like Bi2O2CO3 product is successfully prepared via a simple one-step hydrothermal synthesis, which is spirally assembled by the {001} facet-dominated nanosheets. It is testified that the additive CA plays an important inducing role in forming the chemical composition of Bi2O2CO3, the nanosized sheet-type subunits, and the exposure of the {001} facet, while the NDSA greatly improves the dispersity and porous structure of the Bi2O2CO3 microflower. Due to the nano-size effect and distortion of surface Bi-O bonds, the Bi2O2CO3 microflower could be excited by the visible light to exhibit a superior photocatalytic performance in the degradation of tetracycline (TC). Besides, it is found the exposed {001} facet of Bi2O2CO3 would preferentially generate holes during the illumination process, thus enhancing the photooxidative activity of the Bi2O2CO3 microflower. Finally, the structural and optical features of the Bi2O2CO3 microflower have been discussed in detail, and its photocatalytic mechanism has also been proposed in this work.

13.
Zhongguo Zhong Yao Za Zhi ; 45(2): 405-411, 2020 Jan.
Artigo em Chinês | MEDLINE | ID: mdl-32237325

RESUMO

This work aimed to investigate the intestinal absorption characteristics of Laportea bulbifera extract in normal and rheumatoid arthritis model rats. The contents of neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, rutin, kaempferol-3-O-rutinoside, galuteolin, quercetin and isoquercetin in intestinal absorption solution samples were detected by UPLC-MS/MS with 5.0 g·L~(-1) as the absorption concentration. The cumulative absorption(Q) and absorption rate constant(K_a) were calculated, and the absorption characteristics of different components of L. bulbifera in intestinal absorption solution of normal rats and rheumatoid arthritis rats were compared. The results showed that all the eight index components in the extract of L. bulbifera could be absorbed into the intestinal capsule, the cumulative absorption-time curve of each component showed an upward trend without saturation, and the correlation regression coefficient(R~2) was greater than 0.92, which is consistent with the zero-order absorption rate process. It was speculated that the possible absorption mode of each component was passive diffusion. In normal condition, the absorption of ileum was the best(except chlorogenic acid), and in pathological condition, duodenum was the best. The total absorption of 8 components in each intestinal segment of RA rats was better than that of normal rats, which speculated that rheumatoid arthritis may change the specific site of drug absorption. The experimental results showed that rheumatoid arthritis could change the intestinal absorption of the extract of L. bulbifera, and its mechanism needs further study.


Assuntos
Artrite Reumatoide/tratamento farmacológico , Absorção Intestinal , Extratos Vegetais/uso terapêutico , Urticaceae/química , Animais , Cromatografia Líquida de Alta Pressão , Intestinos/efeitos dos fármacos , Ratos , Espectrometria de Massas em Tandem
14.
Phytopathology ; 110(7): 1319-1325, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-32154765

RESUMO

Poplar Cytospora canker, which is mainly caused by Cytospora chrysosperma, is one of the most destructive and widespread tree diseases worldwide. Although oxalic acid (OA) is demonstrated as an important virulence determinant in several necrotrophic fungi, specific functions of OA during pathogenesis remain controversial. Here, we identified three genes (CcOah, CcOdc1, and CcOdc2) directly involved in OA biosynthesis and catabolism in C. chrysosperma. We demonstrated that CcOah is required for OA biogenesis. All three genes were found to be highly upregulated during early infection stages of the poplar stem. The deletion of any of the three genes led to an obvious reduction of pycnidial production but no abnormality of hyphal growth and morphology. Furthermore, the individual deletion strain exhibited significantly limited lesion sizes on poplar twigs and leaves. Exogenous application of OA or citric acid can complement the virulence defects of ΔCcOah and ΔCcOdc1 strains. We further found that the ΔCcOah strain strongly promoted reactive oxygen species burst of poplar leaves during infection. Finally, induced secretion of OA was observed by monitoring color change of the plates after poplar stem extracts were added in the cultures; however, we failed to quantify OA concentration by high-performance liquid chromatography. Taken together, the present results provide insights into the function of OA acting as an important virulence factor of C. chrysosperma.


Assuntos
Ascomicetos , Populus , Ácido Oxálico , Doenças das Plantas , Virulência
15.
New Phytol ; 226(5): 1461-1479, 2020 06.
Artigo em Inglês | MEDLINE | ID: mdl-32040203

RESUMO

The fungus Verticillium dahliae causes vascular wilt disease on hundreds of plant species. Homologs of the bZIP transcription factor Atf1 are required for virulence in most pathogenic fungi, but the molecular basis for their involvement is largely unknown. We performed targeted gene deletion, expression analysis, biochemistry and pathogenicity assays to demonstrate that VdAtf1 governs pathogenesis via the regulation of nitrosative resistance and nitrogen metabolism in V. dahliae. VdAtf1 controls pathogenesis via the regulation of nitric oxide (NO) resistance and inorganic nitrogen metabolism rather than oxidative resistance and is important for penetration peg formation in V. dahliae. VdAtf1 affects ammonium and nitrate assimilation in response to various nitrogen sources. VdAtf1 may be involved in regulating the expression of VdNut1. VdAtf1 responds to NO stress by strengthening the fungal cell wall, and by causing over-accumulation of methylglyoxal and glycerol, which in turn impacts NO detoxification. We also verified that the VdAtf1 ortholog in Fusarium graminearum mediates nitrogen metabolism, suggesting conservation of this function in related plant pathogenic fungi. Our findings revealed new functions of VdAtf1 in pathogenesis, response to nitrosative stress and nitrogen metabolism in V. dahliae. The results provide novel insights into the regulatory mechanisms of the transcription factor VdAtf1 in virulence.

16.
Xenobiotica ; 50(7): 822-830, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31791186

RESUMO

The study aimed to compare the pharmacokinetic properties of quercitrin, astragalin, afzelin and taxifolin, four major bioactive components of Polygonum orientale inflorescence extracts, between sham-operated and myocardial ischemia-reperfusion injury (MIRI) rats.Rats were divided into two groups: MIRI model and sham-operated. The blood samples were collected according to the time schedule. The levels of quercitrin, astragalin, afzelin and taxifolin in the plasma at designated time points were determined using an HPLC-MS/MS method. Various pharmacokinetic parameters were estimated from the plasma concentration versus time data using non-compartmental methods. After the administration of the Chinese herb Polygonum orientale inflorescence extracts, the Cmax, AUC, as well as MRT, increased, while CL decreased, in MIRI model compared to the sham-operated animals.These results suggest that the pathological damage of ischemia-reperfusion had a significant impact on the pharmacological effects of Polygonum orientale inflorescence extracts on ischemic heart disease.The method had been successfully applied to evaluate the pharmacokinetics of quercitrin, astragalin, afzelin and taxifolin in rat plasma after the oral administration of Chinese herb Polygonum orientale inflorescence extracts in rats.


Assuntos
Medicamentos de Ervas Chinesas/farmacocinética , Extratos Vegetais/farmacocinética , Polygonum , Animais , Quempferóis/metabolismo , Manosídeos/metabolismo , Proantocianidinas/metabolismo , Quercetina/análogos & derivados , Quercetina/metabolismo , Ratos Sprague-Dawley , Traumatismo por Reperfusão
17.
Fungal Genet Biol ; 135: 103289, 2020 02.
Artigo em Inglês | MEDLINE | ID: mdl-31704368

RESUMO

The HOG (high-osmolarity glycerol) pathway is critical for the appropriate adaptation to adverse conditions. Here, we demonstrated that the deletion of CgHog1 resulted in enhanced sensitivity to osmotic stress and increased resistance to fludioxonil in the poplar anthracnose fungus Colletotrichum gloeosporioides. The accumulation of chitin around hyphal tips was obviously decreased in the ΔCgHog1 strain under sorbitol, whereas it strongly was increased in the response to fludioxonil compared with the wild type. To investigate the underlying mechanism of CgHog1-mediated adaption to osmotic stress and fludioxonil, transcriptomic profiles were performed in both the ΔCgHog1 strain and the wild type under the treatment of sorbitol and fludioxonil, respectively. Under sorbitol, genes associated with glycolysis, lipid metabolism, and accumulation of soluble sugars and amino acids were differentially expressed; under fludioxonil, vesicle trafficking-related genes were highly downregulated in the ΔCgHog1 strain, which was consistent with abnormal vacuoles distribution and morphology of hyphae, indicating that the growth defect caused by fludioxonil may be associated with disruption of endocytosis. Taken together, we elucidated the adaptation mechanisms of how CgHog1 regulates appropriate response to sorbitol and fludioxonil via different metabolism pathways. These findings extend our insights into the HOG pathway in fungi.

18.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi ; 35(10): 865-871, 2019 Oct.
Artigo em Chinês | MEDLINE | ID: mdl-31814561

RESUMO

Objective To construct a double transfected Flp-InTM CHO cell line stably expressing both cytochrome P450 family 2 subfamily A member 13(CYP2A13) and multidrug resistance-associated protein 2(MRP2). Methods We constructed the recombinant plasmids of pCMV6-NEO-CYP2A13 and pcDNA5-MRP2. The pCMV6-NEO-CYP2A13 recombinant plasmid was first transfected into Flp-InTM CHO cells, and CYP2A13-Flp-InTM CHO cells with higher CYP2A13 activity were screened using limiting dilution method and 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) cytotoxicity assay. Thereafter, pcDNA5-MRP2 was transfected into CYP2A13-Flp-InTM CHO cells. The expression levels and activities of CYP2A13 and MRP2 in the double transfected cells and normal cells were detected by real-time quantitative PCR, Western blot analysis and NNK cytotoxicity assay in order to screen Flp-InTM CHO cells with stable expression of CYP2A13 and MRP2. Results Compared with non-transfected cells, the expression of CYP2A13 and the sensitivity of NNK toxicity in CYP2A13-Flp-InTM CHO cells increased. The expression of CYP2A13 and MRP2 in CYP2A13/MRP2-Flp-InTM CHO cells also increased significantly. Compared with CYP2A13-Flp-InTM CHO cells, CYP2A13/MRP2-Flp-InTM CHO cells showed no significant difference in CYP2A13 expression; the expression of MRP2 increased while the sensitivity of NNK toxicity decreased significantly. Conclusion The double transfected cell model of CYP2A13 and MRP2 has been successfully established, which lays the foundation for the study of in situ activation of respiratory carcinogens.


Assuntos
Hidrocarboneto de Aril Hidroxilases/genética , Células CHO , Proteínas Associadas à Resistência a Múltiplos Medicamentos/genética , Animais , Carcinógenos/toxicidade , Cricetinae , Cricetulus , Nitrosaminas/toxicidade , Plasmídeos , Transfecção
19.
Artigo em Inglês | MEDLINE | ID: mdl-31772602

RESUMO

Bletilla striata has been widely used as a valuable hemostatic agent for thousands of years due to the high levels of bioactive constituents it contains. Here, we used a sensitive ultrahigh-performance liquid chromatography-tandem mass spectroscopy (UPLC-MS/MS) method for the simultaneous determination of three major active ingredients of the B. striata extract, namely, α-isobutylmalic acid, gymnoside I, and militarine in rat plasma. The three major active ingredients were determined using the multiple reaction monitoring (MRM) mode at m/z 189 ⟶ 129 for α-isobutylmalic acid, m/z 457.2 ⟶ 285.1 for gymnoside I, m/z 725.3 ⟶ 457.2 for militarine, and m/z 417.0 ⟶ 267.0 for the IS puerarin. All calibration curves showed good linearity (R 2 ≥ 0.999) over the concentration range with the lower limit of quantification between 0.015 and 0.029 µg/mL. The relative standard deviations of intraday and interday measurements were less than 15%, and the method was accurate within 93.3-100.4%. The extraction recovery was 92.65-100.98%, and no matrix effect was observed. The results indicated that after oral administration of B. striata in rats, the T max of α-isobutylmalic acid was significantly longer than that of gymnoside I and militarine and the mean residence time and area under the curve of α-isobutylmalic acid and gymnoside I in rats were significantly higher than those of militarine. Moreover, the blood concentration-time curve of α-isobutylmalic acid showed double peaks, suggesting that α-isobutylmalic acid could exhibit the phenomenon of enterohepatic circulation or metabolic conversion. We also explored some of the pharmacokinetic characteristics of three ingredients from B. striata extract in vivo, and the data obtained may provide a basis for the further investigation of B. striata.

20.
Molecules ; 24(21)2019 Oct 27.
Artigo em Inglês | MEDLINE | ID: mdl-31717895

RESUMO

Polygonum capitatum Buch.-Ham. ex D. Don is traditionally used by Hmong for the treatment of urinary tract infections and pyelonephritis. Information regarding the pharmacokinetic behavior of the extract in the condition of pyelonephritis is lacking. In the present study, we aimed to compare the pharmacokinetic properties of gallic acid (GA), protocatechuic acid (PCA), and quercitrin (QR)-the main bioactive constituents in the herb-in normal and pyelonephritis rats. The plasma samples were collected at various time points after administration of a single dose of Polygonum capitatum extract. The plasma level of GA, PCA, and QR at the designed time points was determined by ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) and drug concentration versus time plots were constructed to estimate the pharmacokinetic parameters. The AUC(0-t), AUC(0-∞), MRT(0-t), and CL of GA, PCA, and QR in pyelonephritis rats was significantly different from those of the normal rats. The results indicated that the three constituents have higher rate of uptake and slower rate of elimination in the rats with pyelonephritis, suggesting altered rate and extent of drug metabolism.


Assuntos
Ácido Gálico/farmacocinética , Hidroxibenzoatos/farmacocinética , Extratos Vegetais/uso terapêutico , Polygonum/química , Quercetina/análogos & derivados , Animais , Medicamentos de Ervas Chinesas/uso terapêutico , Feminino , Pielonefrite/tratamento farmacológico , Pielonefrite/metabolismo , Quercetina/farmacocinética , Ratos , Ratos Sprague-Dawley
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