The toxicity of acetaldehyde and age related changes on oxidoreductases in the liver,brain, kidney, and musele of female albino rats (Wistar strain) were studied. The specific activities of lactate [LDH], isocitrate [...The toxicity of acetaldehyde and age related changes on oxidoreductases in the liver,brain, kidney, and musele of female albino rats (Wistar strain) were studied. The specific activities of lactate [LDH], isocitrate [ICDH (NAD/NADP)], succinate [SDH], malate [MDH], glutamate [GDH] and glucose-6-Phosphate [G-6-PDH] dehydrogenases were signifieantly inereased as a function of age. However, acetaldehyde treatment significantly inhibited oxidoreductases in the tissues of 21, 90 and 180 day old rats. Liver enzymes of young (21 days) rats exhibited greater sensitivity to acetaldehyde toxicity. Similar inhibition of oxidoreductases in brain and kidney of adult (180 days) rats treated with acetaldehyde was observed. LDH and GDH as compared to other enzymes studied showed higher susceptibility to acetaldehyde toxicity. The differential sensitivity of tissues and inhibition of oxidoreductases by acetaldehyde as a function of age could be attributed to hypoxic conditions, energy crisis, and mitochondrial structural changes. The results suggest that acetaldehyde affects oxidation of glucose via HMP shuni pathway, glycolytic pathway and Krebs cycle resulting in the impairment of carbohydrate metabolism展开更多
Objective Hepatocellular carcinoma(HCC)is the third leading cause of cancer-associated death worldwide.As a first-line drug for advanced HCC treatment,lenvatinib faces a significant hurdle due to the development of bo...Objective Hepatocellular carcinoma(HCC)is the third leading cause of cancer-associated death worldwide.As a first-line drug for advanced HCC treatment,lenvatinib faces a significant hurdle due to the development of both intrinsic and acquired resistance among patients,and the underlying mechanism remains largely unknown.The present study aims to identify the pivotal gene responsible for lenvatinib resistance in HCC,explore the potential molecular mechanism,and propose combinatorial therapeutic targets for HCC management.Methods Cell viability and colony formation assays were conducted to evaluate the sensitivity of cells to lenvatinib and dicoumarol.RNA-Seq was used to determine the differences in transcriptome between parental cells and lenvatinib-resistant(LR)cells.The upregulated genes were analyzed by GO and KEGG analyses.Then,qPCR and Western blotting were employed to determine the relative gene expression levels.Afterwards,the intracellular reactive oxygen species(ROS)and apoptosis were detected by flow cytometry.Results PLC-LR and Hep3B-LR were established.There was a total of 116 significantly upregulated genes common to both LR cell lines.The GO and KEGG analyses indicated that these genes were involved in oxidoreductase and dehydrogenase activities,and reactive oxygen species pathways.Notably,NAD(P)H:quinone oxidoreductase 1(NQO1)was highly expressed in LR cells,and was involved in the lenvatinib resistance.The high expression of NQO1 decreased the production of ROS induced by lenvatinib,and subsequently suppressed the apoptosis.The combination of lenvatinib and NQO1 inhibitor,dicoumarol,reversed the resistance of LR cells.Conclusion The high NQO1 expression in HCC cells impedes the lenvatinib-induced apoptosis by regulating the ROS levels,thereby promoting lenvatinib resistance in HCC cells.展开更多
Azoreductases are involved in the bioremediation by bacteria of azo dyes found in waste water.In the gut flora,they activate azo pro-drugs,which are used for treatment of inflammatory bowel disease,releasing the activ...Azoreductases are involved in the bioremediation by bacteria of azo dyes found in waste water.In the gut flora,they activate azo pro-drugs,which are used for treatment of inflammatory bowel disease,releasing the active component 5-aminosalycilic acid.The bacterium P.aeruginosa has three azoreductase genes,paAzoR1,paAzoR2 and paAzoR3,which as recombinant enzymes have been shown to have different substrate specificities.The mechanism of azoreduction relies upon tautomerisation of the substrate to the hydrazone form.We report here the characterization of the P.aeruginosa azoreductase enzymes,including determining their thermostability,cofactor preference and kinetic constants against a range of their favoured substrates.The expression levels of these enzymes during growth of P.aeruginosa are altered by the presence of azo substrates.It is shown that enzymes that were originally described as azoreductases,are likely to act as NADH quinone oxidoreductases.The low sequence identities observed among NAD(P)H quinone oxidoreductase and azoreductase enzymes suggests convergent evolution.展开更多
Background:Aldo-keto oxidoreductase(AKR)inhibitors could reverse the resistance of several cancer cells to cis-platin,but their role in resistance remains unclear.Methods:We verified the difference of AKR1Cs expressio...Background:Aldo-keto oxidoreductase(AKR)inhibitors could reverse the resistance of several cancer cells to cis-platin,but their role in resistance remains unclear.Methods:We verified the difference of AKR1Cs expression by Western blot,RNA sequencing and qRT-PCR.The differences of AKR1Cs expression were analyzed and inferred.Use Assay of NADH and NAD^(+)content to verify the inference.The Docking experience was used to verify the affinity between MPA,MCFLA,MLS and AKR1C3.Results:Our RNA-seq results showed de novo NAD biosynthesis-related genes and NAD(P)H-dependent oxidoreductases were significantly upregulated in cis-platin-resistant HepG2 hepatic cancer cells(HepG2-RC cells)compared with HepG2 cells.At least 63 NAD(P)H-dependent reductase/oxidases were upregulated in HepG2-RC cells at least twofold.Knockdown of AKR1Cs could increase cis-platin sensitivity in HepG2-RC cells about two-fold.Interestingly,the AKR1C inhibitor meclofenamic acid could increase the cis-platin sensitivity of HepG2-RC cells about eight-fold,indicating that the knockdown of AKR1Cs only partially reversed the resistance.Meanwhile,the amount of total NAD and the ratio of NADH/NAD^(+)were increased in HepG2-RC cells compared with HepG2 cells.The ratio of NADH/NAD^(+)in HepG2-RC cells was almost seven-fold higher than in HepG2 or HL-7702 cells.Increased NADH expression could be explained as a directly operating antioxidant to scavenge cis-platin-induced radicals.Conclusion:We report here that NADH,which is produced by NAD(P)Hdependent oxidoreductases,plays a key role in the AKR-associated cis-platin resistance of HepG2 hepatic cancer cells.展开更多
The energy shift toward glycolysis is one of the hallmarks of cancer.Complex I is a vital enzyme complex necessary for oxidative phosphorylation.The mitochondrially encoded NADH:ubiquinone oxidoreductase core subunit ...The energy shift toward glycolysis is one of the hallmarks of cancer.Complex I is a vital enzyme complex necessary for oxidative phosphorylation.The mitochondrially encoded NADH:ubiquinone oxidoreductase core subunit 1(MT-ND1)is the largest subunit coded by mitochondria of complex I.The present study summarizes the structure and biological function of MT-ND1.From databases and literature,the expressions and mutations of MT-ND1 in a variety of cancers have been reviewed.MT-ND1 may be a biomarker for cancer diagnosis and prognosis.It is also a potential target for cancer therapy.展开更多
Many approaches to neurodegenerative diseases that focus on amyloid-βclearance and gene therapy have not been successful.Some therapeutic applications focus on enhancing neuronal cell survival during the pathogenesis...Many approaches to neurodegenerative diseases that focus on amyloid-βclearance and gene therapy have not been successful.Some therapeutic applications focus on enhancing neuronal cell survival during the pathogenesis of neurodegenerative diseases,including mitochondrial dysfunction.Plasma membrane(PM)redox enzymes are crucial in maintaining cellular physiology and redox homeostasis in response to mitochondrial dysfunction.Neurohormetic phytochemicals are known to induce the expression of detoxifying enzymes under stress conditions.In this study,mechanisms of neuroprotective effects of 4-hydroxycinnamic acid(HCA)were examined by analyzing cell survival,levels of abnormal proteins,and mitochondrial functions in two different neuronal cells.HCA protected two neuronal cells exhibited high expression of PM redox enzymes and the consequent increase in the NAD^(+)/NADH ratio.Cells cultured with HCA showed delayed apoptosis and decreased oxidative/nitrative damage accompanied by decreased ROS production in the mitochondria.HCA increased the mitochondrial complexes I and II activities and ATP production.Also,HCA increased mitochondrial fusion and decreased mitochondrial fission.Overall,HCA maintains redox homeostasis and energy metabolism under oxidative/metabolic stress conditions.These findings suggest that HCA could be a promising therapeutic approach for neurodegenerative diseases.展开更多
AIM:To determine whether the microRNA-27b-3p(miR-27b-3p)/NF-E2-related factor 2(Nrf2)pathway plays a role in human retinal pigment epithelial(hRPE)cell response to high glucose,how miR-27b-3p and Nrf2 expression are r...AIM:To determine whether the microRNA-27b-3p(miR-27b-3p)/NF-E2-related factor 2(Nrf2)pathway plays a role in human retinal pigment epithelial(hRPE)cell response to high glucose,how miR-27b-3p and Nrf2 expression are regulated,and whether this pathway could be specifically targeted.METHODS:hRPE cells were cultured in normal glucose or high glucose for 1,3,or 6d before measuring cellular proliferation rates using cell counting kit-8 and reactive oxygen species(ROS)levels using a dihydroethidium kit.miR-27b-3p,Nrf2,NAD(P)H quinone oxidoreductase 1(NQO1)and heme oxygenase-1(HO-1)mRNA and protein levels were analyzed using reverse transcription quantitative polymerase chain reaction(RT-qPCR)and immunocytofluorescence(ICF),respectively.Western blot analyses were performed to determine nuclear and total Nrf2 protein levels.Nrf2,NQO1,and HO-1 expression levels by RT-qPCR,ICF,or Western blot were further tested after miR-27b-3p overexpression or inhibitor lentiviral transfection.Finally,the expression level of those target genes was analyzed after treating hRPE cells with pyridoxamine.RESULTS:Persistent exposure to high glucose gradually suppressed hRPE Nrf2,NQO1,and HO-1 mRNA and protein levels and increased miR-27b-3p mRNA levels.High glucose also promoted ROS release and inhibited cellular proliferation.Nrf2,NQO1,and HO-1 mRNA levels decreased after miR-27b-3p overexpression and,conversely,both mRNA and protein levels increased after expressing a miR-27b-3p inhibitor.After treating hRPE cells exposed to high glucose with pyridoxamine,ROS levels tended to decreased,proliferation rate increased,Nrf2,NQO1,and HO-1 mRNA and protein levels were upregulated,and miR-27b-3p mRNA levels were suppressed.CONCLUSION:Nrf2 is a downstream target of miR-27b-3p.Furthermore,the miR-27b-3p inhibitor pyridoxamine can alleviate high glucose injury by regulating the miR-27b-3p/Nrf2 axis.展开更多
Aim: Prostane, a polyherbal formulation, was evaluated for its efficacy on 5α-reductase inhibition, α-adrenergic anta-gonistic activity and testosterone-induced prostatic hyperplasia. Methods: 5α-reductase inhibiti...Aim: Prostane, a polyherbal formulation, was evaluated for its efficacy on 5α-reductase inhibition, α-adrenergic anta-gonistic activity and testosterone-induced prostatic hyperplasia. Methods: 5α-reductase inhibition was evaluated usingrat prostate homogenate as an enzyme source. Adrenergic antagonistic activity was evaluated using isolated rat vas def-erens. Experimental prostatic hyperplasia was induced in rats by giving testosterone 3 rog/kg sc for 21 days. Re-sults: Prostane dose-dependently inhibited 5α-reductase activity and exhibited α-adrenergic antagonistic activity. Treat-ment with Prostane at 250, 500 and 750 mg/kg body wt, po for 21 days significantly reduced the prostatic weight, theepithelial height and the stromal proliferation in experimental prostatic hypertrophy. Conclusion: Prostane is effectivein the treatment of experimental prostatic hypertrophy in rats and may be passed on to clinical trials on benign prostatichypertrophy after necessary toxicological evaluations. (Asian J Androl 1999 Dec; 1: 175-179)展开更多
Objective Localization of the glutathione dependent Nitroblue tetrazolium (NBT) reductase in fresh frozen sections of mouse skin and possible dependence of NBT reductase on tissue thiol levels has been investigated. ...Objective Localization of the glutathione dependent Nitroblue tetrazolium (NBT) reductase in fresh frozen sections of mouse skin and possible dependence of NBT reductase on tissue thiol levels has been investigated. Methods The fresh frozen tissue sections (8m thickness) were prepared and incubated in medium containing NBT, reduced glutathione (GSH) and phosphate buffer. The staining for GSH was performed with mercury orange. Results The activity of the NBT-reductase in mouse skin has been found to be localized in the areas rich in glutathione and actively proliferating area of the skin. Conclusion The activity of the NBT-reductase seems to be dependent on the glutathione contents.展开更多
AIM:To investigate the effects of blueberry on hepatic fibrosis and NF-E2-related factor 2(Nrf2) transcription factor in rats.METHODS:Forty-five male Sprague-Dawley rats were randomly divided into control group(A);CCl...AIM:To investigate the effects of blueberry on hepatic fibrosis and NF-E2-related factor 2(Nrf2) transcription factor in rats.METHODS:Forty-five male Sprague-Dawley rats were randomly divided into control group(A);CCl4-induced hepatic fibrosis group(B);blueberry prevention group(C);Dan-shao-hua-xian capsule(DSHX) prevention group(D);and blueberry + DSHX prevention group(E).Liver fibrosis was induced in rats by subcutaneous injection of CCl4 and a high-lipid/low-protein diet for 8 wk(except the control group).The level of hyaluronic acid(HA) and alanine aminotransferase(ALT) in serum was examined.The activity of superoxide dismutase(SOD),glutathione-S-transferase(GST) and malondialdehyde(MDA) in liver homogenates was determined.The degree of hepatic fibrosis was evaluated by hematoxylin and eosin and Masson staining.Expression of Nrf2 and NADPH quinone oxidoreductase 1(Nqo1) was detected by real-time reversed transcribed-polymerase chain reaction,immunohistochemical techniques,and western blotting.RESULTS:Compared with group B,liver indices,levels of serum HA and ALT of groups C,D and E were reduced(liver indices:0.038 ± 0.008,0.036 ± 0.007,0.036 ± 0.005 vs 0.054 ± 0.009,P<0.05;HA:502.33 ± 110.57 ng/mL,524.25 ± 255.42 ng/mL,499.25 ± 198.10 ng/mL vs 828.50 ± 237.83 ng/mL,P<0.05;ALT:149.44 ± 16.51 U/L,136.88 ± 10.07 U/L,127.38 ± 11.03 U/L vs 203.25 ± 31.62 U/L,P<0.05),and SOD level was significantly higher,but MDA level was lower,in liver homogenates(SOD:1.36 ± 0.09 U/mg,1.42 ± 0.13 U/mg,1.50 ± 0.15 U/mg vs 1.08 ± 0.19 U/mg,P<0.05;MDA:0.294 ± 0.026 nmol/mg,0.285 ± 0.025 nmol/mg,0.284 ± 0.028 nmol/mg vs 0.335 ± 0.056 nmol/mg,P<0.05).Meanwhile,the stage of hepatic fibrosis was significantly weakened(P<0.05).Compared with group A,the activity of GST liver homogenates and expression levels of Nrf2 and Nqo1 in group B were elevated(P<0.05).The expression level of Nrf2 and Nqo1 in groups C,D,and E were increased as compared with group B,but the difference was not significant.CONCLUSION:Blueberry has preventive and protective effects on CCl4-induced hepatic fibrosis by reducing hepatocyte injury and lipid peroxidation.However,these effects may not be related to the activation of Nrf2 during long-term of CCl4.展开更多
No study has been performed on identifying microRNAs(miRNAs) and their targets in cotton although cotton is one of the most important fiber and economic crops around the world.In this study,
In order to improve beef color and color stability, step-chilling (SC) was applied on excised bovine Iongissimus lumborum muscle, with chilling starting at 0-4℃ for 5 h, then holding the temperature at 12-18℃ for ...In order to improve beef color and color stability, step-chilling (SC) was applied on excised bovine Iongissimus lumborum muscle, with chilling starting at 0-4℃ for 5 h, then holding the temperature at 12-18℃ for 6 h, followed by 0-4℃ again until 24 h post-mortem, pH and temperature were measured during rigor on SC loins as well as those subjected to routine chilling (RC, 0-4℃, till 24 h post-mortem). Color L*, a*, b* values, metmyoglobin (MetMb) content, MetMb reducing ability (MRA) and NADH content were determined on samples aged for 1, 7, and 14 d. Sarcoplasmic proteome analysis was only conducted on d 1 samples. The results showed muscles subjected to SC maintained a temperature at around 15℃ for 5 to 10 h post-mortem, and exhibited a slow temperature decline, but rapid pH decline. Beef steaks treated with SC had higher L*, a*, b* and chroma values than those of RC samples at 1 and 7 d chilled storage (0-4℃), while showing no significant difference for a*, b* and chroma values at d 14. The SC samples also exhibited a lower relative content of surface MetMb, higher MRA and NADH content, compared with RC beef steaks during storage, indicating the SC-treated beef showed an improved color stability. Eleven differential protein spots/nine proteins were identified by two-dimensional gel electrophoresis and mass spectrometry, and those proteins were mainly involved in redox, chaperone binding, metabolic and peroxidase activity. Oxidoreductases play a role in decreasing the oxidation-induced myoglobin oxidation and benefiting the production of NADH, and finally improving the colour of beef. Of these, pyruvate dehydrogenase E1 component subunit beta showed a positive correlation with color L*, a*, b* values and accounted for more than 60% of the variation in color values; this protein can be considered as a potential beef color biomarker. The present study provided valuable information for studies on the molecular mechanism of color improvement from step-chilling, as well as for identifying markers associated with beef color.展开更多
The WW domain-containing oxidoreductase(WWOX) is a tumor suppressor in a variety of cancers, including breast cancer. Reduced WWOX expression is associated with the basal-like subtype and a relatively poor disease-f...The WW domain-containing oxidoreductase(WWOX) is a tumor suppressor in a variety of cancers, including breast cancer. Reduced WWOX expression is associated with the basal-like subtype and a relatively poor disease-free survival rate among breast cancer patients. Though several WWOX partners have been identified, the functional mechanisms of WWOX's role in cancers have not been fully addressed to date. In the current study, we found WWOX suppresses expression of KLF5—an oncogenic transcription factor—at protein level, and suppresses cancer cell proliferation in both bladder and breast cancer cell lines. Furthermore, we demonstrated that WWOX physically interacts with KLF5 via the former's WW domains and the latter's PY motifs. Interestingly, we found the expression of WWOX negatively correlates with KLF5 expression in a panel of breast cancer cell lines. Taken together, we conjecture that WWOX may suppress cancer cell proliferation partially by reducing the expression of KLF5.展开更多
Objective: To investigate the relationship between NAD(P)H:quinone oxidoreductase 1 (NQOI) C609T polymorphism and colon cancer risk in farmers from western region of Inner Mongolia. Methods: Polymerase chain re...Objective: To investigate the relationship between NAD(P)H:quinone oxidoreductase 1 (NQOI) C609T polymorphism and colon cancer risk in farmers from western region of Inner Mongolia. Methods: Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was performed to analyze NQO1 C609T polymorphism from 160 healthy controls and 76 colon cancer patients. Results: Among the colon cancer patients, the incidence of NQOI T allele (53.29%) was significantly higher than it in control group (33.75%, P〈0.001). The individuals with NQO1 T allele had higher risk [2.239 (95% CI: 1.510-3.321) times] to develop colon cancer than individuals with NQO1 C allele. The incidence of NQO1 (TFI-) (34.21%) in colon cancer patients was higher than that in control group (15.62%, P〈0.001). Odds ratios (OR) analysis suggested that NQO1 (T/F) and NQOI (T/C) genotype carriers had 3.813 (95% CI: 1.836-7.920) times and 2.080 (1.026-4.219) times risk compared with wild-type NQO1 (C/C) gene carriers in developing colon cancer. Individuals with NQO1 (T/I') genotype had 2.541 (95% CI: 0.990-6.552) times, 3.713 (95% CI: 1.542-8.935) times, and 3.471 (95% CI: 1.356-8.886) times risk than individuals with NQOI (T/C) or NQOI (C/C) genotype in well- differentiated, moderately-differentiated, and poorly-differentiated colon cancer patients, respectively. Conclusions: NQO1 gene C609T could be one of risk factors of colon cancer in farmers from western region of Inner Mongolia,展开更多
Phytochromes in rice are encoded by a gene family composed of three members, PHYA, PHYB, and PHYC. Through characterizing the phytochrome mutants and wild type (WT) in terms of photomorphogenesis, roles of individua...Phytochromes in rice are encoded by a gene family composed of three members, PHYA, PHYB, and PHYC. Through characterizing the phytochrome mutants and wild type (WT) in terms of photomorphogenesis, roles of individual phytochromes have been preliminarily explored in regulating rice de-etiolation, flowering time and fertility. However, little information has been reported about whether or how phytochromes affect chlorophyll biosynthesis and chloroplast development in rice. In this study, we compared the chlorophyll contents of wild type and the phyA, phyB and phyAphyB mutants grown under either white light (WL) or red light (R). The results suggest that phyB perceives R to positively regulate chlorophyll biosynthesis, while the role of phyA can be detected only in the phyB-deficient mutant. Analyses of the expression levels of genes involved in chlorophyll biosynthesis revealed that phytochromes affected the chlorophyll biosynthesis by regulating protochlorophyll oxidoreductase A (PORA) expression. The role of phyB in chloroplast development was also analyzed, and the results suggest that phyB perceives R to regulate chloroplast development by affecting the numbers of chloroplasts and grana, as well as the chloroplast membrane system.展开更多
CO2 fixation pathway of Caminibacter profundus, a chemolithoautotrophic e-Proteobacteria from deep-sea hydrothermal vent, was determined and characterized by genetic and enzymatic analyses. Gene expression of key enzy...CO2 fixation pathway of Caminibacter profundus, a chemolithoautotrophic e-Proteobacteria from deep-sea hydrothermal vent, was determined and characterized by genetic and enzymatic analyses. Gene expression of key enzymes for CO2 fixation in response to salinity, pH and O2 in Medium 829 were also investigated. The results demonstrate that C. profundus contained aclB, porA and oorA, the genes encoding key enzymes of reductive tricarboxylic acid (rTCA) cycle. However, genes fragments of cbbL and cbbMencoding key enzyme of Calvin cycle were not recovered. Key enzymatic activities of ATP citrate lyase (ACL), pyruvate: ferredoxin oxidoreductase (POR) and 2-oxoglutarate: ferredoxin oxidoreductase (OOR) were also present in C. profun- dus. The combination of genetic and enzymatic analyses confirm that C. profundus adopted rTCA cycle for carbon assimilation. The results of aclB and oorA relative expressions of C. profundus demonstrate that the ranges of environmental factors for high genes expression were sea salt 3.0%-5.0% (optimum 3.0%), pH 5.0-6.5(optimum pH 6.5), anaerobic to microaerobic conditions (optimum 1.0% 02). Gene expression pat- terns under different conditions show similar patterns with bacterial growth, revealing that key rTCA cycle genes provided molecular basis for bacterial growth and propagation. Our results suggest that C. profun- dus could regulate key genes of rTCA cycle for carbon assimilation and energy metabolism in response to environmental fluctuations in hydrothermal vent.展开更多
The present study aimed to explore the molecular mechanisms underlying the increase of nicotinamide adenine dinucleotide phosphate:quinine oxidoreductase 1(NQO1)and y-glutamylcysteine synthetase(γ-GCS)in brain tissue...The present study aimed to explore the molecular mechanisms underlying the increase of nicotinamide adenine dinucleotide phosphate:quinine oxidoreductase 1(NQO1)and y-glutamylcysteine synthetase(γ-GCS)in brain tissues after intracerebral hemorrhage(ICH).The microglial cells obtained from newborn rats were cultured and then randomly divided into the normal control group(NC group),model control group(MC group),rosiglitazone(RSG)intervention group(RSG group),retinoic-acid intervention group(RSG+RA group),and sulfbraphane group(RSG+SF group).The expression levels of NQO1,γ-GCS,and nuclear factor E2-related factor 2(Nrf2)were measured by real-time polymerase chain reaction(RT-PCR)and Western blotting,respectively.The results showed that the levels of NQO1,γ-GCS and Nrf2 were significantly increased in the MC group and the RSG group as compared with those in the NC group(P<0.01).They were found to be markedly decreased in the RSG+RA group and increased in the RSG+SF group when compared with those in the MC group or the RSG group(P<0.01).The RSG+SF group displayed the highest levels of NQO1,γ-GCS,and Nrf2 among the five groups.In conclusion,a medium dose of RSG increased the anti-oxidative ability of thrombinactivated microglia by increasing the expression of NQO1 and γ-GCS.The molecular mechanisms underlying the increase of NQO1 and γ-GCS in thrombin-activated microglia may be associated with the activation of Nrf2.展开更多
BACKGROUND Quinine oxidoreductase 1(NQO1)plays a vital role in protecting normal cells against oxidative damage and electrophilic attack.It is highly expressed in many solid tumors,suggesting a role in cancer developm...BACKGROUND Quinine oxidoreductase 1(NQO1)plays a vital role in protecting normal cells against oxidative damage and electrophilic attack.It is highly expressed in many solid tumors,suggesting a role in cancer development and progression.However,the role of NQO1 in gastric cancer and its effect on cancer development and prognosis have not been fully investigated.AIM To investigate the clinical relevance of NQO1 protein expression in gastric cancer and to explore the potential of NQO1 to serve as a prognostic biomarker and therapeutic target.METHODS In this retrospective study,gastric cancer specimens of 175 patients who were treated between 1995 and 2011 were subjected to immunohistochemistry analyses for NQO1.The correlation of NQO1 expression with gastric cancer prognosis and clinical and pathological parameters was investigated.RESULTS NQO1 protein was overexpressed in 59.43%(104/175)of the analyzed samples.Overexpression of NQO1 was associated with a significantly inferior prognosis.In addition,multivariate analysis suggested that NQO1 overexpression,along with tumor stage and patient age,are prominent prognostic biomarkers for gastric cancer.Moreover,NQO1 overexpression was correlated to a better response to 5-fluorouracil(5-FU)-based adjuvant chemotherapy.CONCLUSION NQO1 overexpression is associated with a significantly poor prognosis and better response to 5-FU in patients with gastric cancer.These findings are relevant for improving therapeutic approaches for gastric cancer patients.展开更多
The growth-inhibiting and apoptosis-inducing effects of WW domain-containing oxidoreductase(WWOX) gene on ovarian cancer cell line A2780 were investigated.The full length cDNA of human WWOX gene was amplified from nor...The growth-inhibiting and apoptosis-inducing effects of WW domain-containing oxidoreductase(WWOX) gene on ovarian cancer cell line A2780 were investigated.The full length cDNA of human WWOX gene was amplified from normal human ovary tissues.The correct cDNA of full length WWOX was subcloned into eukaryocytic expression vector pCMV.After introduction of WWOX gene into cancer cells with liposome,the WWOX mRNA and protein level in the cancer cells were detected by reverse transcription polymerase chain reaction(RT-PCR) and immunoblotting.The growth activities of cancer cells were detected by Trypan blue staining.The clone formation assay in soft agar was employed to observe the proliferation of the cancer cells.Apoptosis was examined by DNA ladder and acridine orange-ethidium bromide fluorescent staining.The results showed that 72 h after WWOX gene transfection,the WWOX expression was increased significantly(P<0.01).The growth of ovarian cancer cells was decreased by 16.41% to 38.49%(P<0.01).The clone formation abilities were reduced(P<0.01).Some cancer cells presented the characteristic morphological changes of apoptosis with obvious ladder bands on electrophoresis.The apoptosis rate was(20.7±6.0)%(P<0.01).It was concluded that over-expression of WWOX gene could induce apoptosis and inhibit the growth of ovarian cancer cells,which might be potentially useful in the gene therapy of ovarian cancers.展开更多
文摘The toxicity of acetaldehyde and age related changes on oxidoreductases in the liver,brain, kidney, and musele of female albino rats (Wistar strain) were studied. The specific activities of lactate [LDH], isocitrate [ICDH (NAD/NADP)], succinate [SDH], malate [MDH], glutamate [GDH] and glucose-6-Phosphate [G-6-PDH] dehydrogenases were signifieantly inereased as a function of age. However, acetaldehyde treatment significantly inhibited oxidoreductases in the tissues of 21, 90 and 180 day old rats. Liver enzymes of young (21 days) rats exhibited greater sensitivity to acetaldehyde toxicity. Similar inhibition of oxidoreductases in brain and kidney of adult (180 days) rats treated with acetaldehyde was observed. LDH and GDH as compared to other enzymes studied showed higher susceptibility to acetaldehyde toxicity. The differential sensitivity of tissues and inhibition of oxidoreductases by acetaldehyde as a function of age could be attributed to hypoxic conditions, energy crisis, and mitochondrial structural changes. The results suggest that acetaldehyde affects oxidation of glucose via HMP shuni pathway, glycolytic pathway and Krebs cycle resulting in the impairment of carbohydrate metabolism
基金supported by the Global Select Project(No.DJK-LX-2022001)of the Institute of Health and Medicine,Hefei Comprehensive National Science Center.
文摘Objective Hepatocellular carcinoma(HCC)is the third leading cause of cancer-associated death worldwide.As a first-line drug for advanced HCC treatment,lenvatinib faces a significant hurdle due to the development of both intrinsic and acquired resistance among patients,and the underlying mechanism remains largely unknown.The present study aims to identify the pivotal gene responsible for lenvatinib resistance in HCC,explore the potential molecular mechanism,and propose combinatorial therapeutic targets for HCC management.Methods Cell viability and colony formation assays were conducted to evaluate the sensitivity of cells to lenvatinib and dicoumarol.RNA-Seq was used to determine the differences in transcriptome between parental cells and lenvatinib-resistant(LR)cells.The upregulated genes were analyzed by GO and KEGG analyses.Then,qPCR and Western blotting were employed to determine the relative gene expression levels.Afterwards,the intracellular reactive oxygen species(ROS)and apoptosis were detected by flow cytometry.Results PLC-LR and Hep3B-LR were established.There was a total of 116 significantly upregulated genes common to both LR cell lines.The GO and KEGG analyses indicated that these genes were involved in oxidoreductase and dehydrogenase activities,and reactive oxygen species pathways.Notably,NAD(P)H:quinone oxidoreductase 1(NQO1)was highly expressed in LR cells,and was involved in the lenvatinib resistance.The high expression of NQO1 decreased the production of ROS induced by lenvatinib,and subsequently suppressed the apoptosis.The combination of lenvatinib and NQO1 inhibitor,dicoumarol,reversed the resistance of LR cells.Conclusion The high NQO1 expression in HCC cells impedes the lenvatinib-induced apoptosis by regulating the ROS levels,thereby promoting lenvatinib resistance in HCC cells.
文摘Azoreductases are involved in the bioremediation by bacteria of azo dyes found in waste water.In the gut flora,they activate azo pro-drugs,which are used for treatment of inflammatory bowel disease,releasing the active component 5-aminosalycilic acid.The bacterium P.aeruginosa has three azoreductase genes,paAzoR1,paAzoR2 and paAzoR3,which as recombinant enzymes have been shown to have different substrate specificities.The mechanism of azoreduction relies upon tautomerisation of the substrate to the hydrazone form.We report here the characterization of the P.aeruginosa azoreductase enzymes,including determining their thermostability,cofactor preference and kinetic constants against a range of their favoured substrates.The expression levels of these enzymes during growth of P.aeruginosa are altered by the presence of azo substrates.It is shown that enzymes that were originally described as azoreductases,are likely to act as NADH quinone oxidoreductases.The low sequence identities observed among NAD(P)H quinone oxidoreductase and azoreductase enzymes suggests convergent evolution.
基金supported by the Science and Technology Development Plan Project of Jilin Province,China[20200708101YY]The Foundation of Jilin Province Science and Technology Department[20200801062GH].
文摘Background:Aldo-keto oxidoreductase(AKR)inhibitors could reverse the resistance of several cancer cells to cis-platin,but their role in resistance remains unclear.Methods:We verified the difference of AKR1Cs expression by Western blot,RNA sequencing and qRT-PCR.The differences of AKR1Cs expression were analyzed and inferred.Use Assay of NADH and NAD^(+)content to verify the inference.The Docking experience was used to verify the affinity between MPA,MCFLA,MLS and AKR1C3.Results:Our RNA-seq results showed de novo NAD biosynthesis-related genes and NAD(P)H-dependent oxidoreductases were significantly upregulated in cis-platin-resistant HepG2 hepatic cancer cells(HepG2-RC cells)compared with HepG2 cells.At least 63 NAD(P)H-dependent reductase/oxidases were upregulated in HepG2-RC cells at least twofold.Knockdown of AKR1Cs could increase cis-platin sensitivity in HepG2-RC cells about two-fold.Interestingly,the AKR1C inhibitor meclofenamic acid could increase the cis-platin sensitivity of HepG2-RC cells about eight-fold,indicating that the knockdown of AKR1Cs only partially reversed the resistance.Meanwhile,the amount of total NAD and the ratio of NADH/NAD^(+)were increased in HepG2-RC cells compared with HepG2 cells.The ratio of NADH/NAD^(+)in HepG2-RC cells was almost seven-fold higher than in HepG2 or HL-7702 cells.Increased NADH expression could be explained as a directly operating antioxidant to scavenge cis-platin-induced radicals.Conclusion:We report here that NADH,which is produced by NAD(P)Hdependent oxidoreductases,plays a key role in the AKR-associated cis-platin resistance of HepG2 hepatic cancer cells.
基金supported by National Natural Science Foundation of China(No.82203232)Scientific Instrument Field Project of Shanghai Science and Technology Commission(No.22142202700)Shanghai Rising-Star Program(No.19QB1404700).
文摘The energy shift toward glycolysis is one of the hallmarks of cancer.Complex I is a vital enzyme complex necessary for oxidative phosphorylation.The mitochondrially encoded NADH:ubiquinone oxidoreductase core subunit 1(MT-ND1)is the largest subunit coded by mitochondria of complex I.The present study summarizes the structure and biological function of MT-ND1.From databases and literature,the expressions and mutations of MT-ND1 in a variety of cancers have been reviewed.MT-ND1 may be a biomarker for cancer diagnosis and prognosis.It is also a potential target for cancer therapy.
基金supported by the National Research Foundation of Korea(NRF)of the Korean Government(NRF-2021R1F1A1051212)by Logsynk Co.Ltd.(2-2021-1435-001).
文摘Many approaches to neurodegenerative diseases that focus on amyloid-βclearance and gene therapy have not been successful.Some therapeutic applications focus on enhancing neuronal cell survival during the pathogenesis of neurodegenerative diseases,including mitochondrial dysfunction.Plasma membrane(PM)redox enzymes are crucial in maintaining cellular physiology and redox homeostasis in response to mitochondrial dysfunction.Neurohormetic phytochemicals are known to induce the expression of detoxifying enzymes under stress conditions.In this study,mechanisms of neuroprotective effects of 4-hydroxycinnamic acid(HCA)were examined by analyzing cell survival,levels of abnormal proteins,and mitochondrial functions in two different neuronal cells.HCA protected two neuronal cells exhibited high expression of PM redox enzymes and the consequent increase in the NAD^(+)/NADH ratio.Cells cultured with HCA showed delayed apoptosis and decreased oxidative/nitrative damage accompanied by decreased ROS production in the mitochondria.HCA increased the mitochondrial complexes I and II activities and ATP production.Also,HCA increased mitochondrial fusion and decreased mitochondrial fission.Overall,HCA maintains redox homeostasis and energy metabolism under oxidative/metabolic stress conditions.These findings suggest that HCA could be a promising therapeutic approach for neurodegenerative diseases.
基金Supported by National Natural Science Foundation of China(No.2020J01652)the Training Project for Young and Middleaged Core Talents in Health System of Fujian Province(No.2016-ZQN-62).
文摘AIM:To determine whether the microRNA-27b-3p(miR-27b-3p)/NF-E2-related factor 2(Nrf2)pathway plays a role in human retinal pigment epithelial(hRPE)cell response to high glucose,how miR-27b-3p and Nrf2 expression are regulated,and whether this pathway could be specifically targeted.METHODS:hRPE cells were cultured in normal glucose or high glucose for 1,3,or 6d before measuring cellular proliferation rates using cell counting kit-8 and reactive oxygen species(ROS)levels using a dihydroethidium kit.miR-27b-3p,Nrf2,NAD(P)H quinone oxidoreductase 1(NQO1)and heme oxygenase-1(HO-1)mRNA and protein levels were analyzed using reverse transcription quantitative polymerase chain reaction(RT-qPCR)and immunocytofluorescence(ICF),respectively.Western blot analyses were performed to determine nuclear and total Nrf2 protein levels.Nrf2,NQO1,and HO-1 expression levels by RT-qPCR,ICF,or Western blot were further tested after miR-27b-3p overexpression or inhibitor lentiviral transfection.Finally,the expression level of those target genes was analyzed after treating hRPE cells with pyridoxamine.RESULTS:Persistent exposure to high glucose gradually suppressed hRPE Nrf2,NQO1,and HO-1 mRNA and protein levels and increased miR-27b-3p mRNA levels.High glucose also promoted ROS release and inhibited cellular proliferation.Nrf2,NQO1,and HO-1 mRNA levels decreased after miR-27b-3p overexpression and,conversely,both mRNA and protein levels increased after expressing a miR-27b-3p inhibitor.After treating hRPE cells exposed to high glucose with pyridoxamine,ROS levels tended to decreased,proliferation rate increased,Nrf2,NQO1,and HO-1 mRNA and protein levels were upregulated,and miR-27b-3p mRNA levels were suppressed.CONCLUSION:Nrf2 is a downstream target of miR-27b-3p.Furthermore,the miR-27b-3p inhibitor pyridoxamine can alleviate high glucose injury by regulating the miR-27b-3p/Nrf2 axis.
文摘Aim: Prostane, a polyherbal formulation, was evaluated for its efficacy on 5α-reductase inhibition, α-adrenergic anta-gonistic activity and testosterone-induced prostatic hyperplasia. Methods: 5α-reductase inhibition was evaluated usingrat prostate homogenate as an enzyme source. Adrenergic antagonistic activity was evaluated using isolated rat vas def-erens. Experimental prostatic hyperplasia was induced in rats by giving testosterone 3 rog/kg sc for 21 days. Re-sults: Prostane dose-dependently inhibited 5α-reductase activity and exhibited α-adrenergic antagonistic activity. Treat-ment with Prostane at 250, 500 and 750 mg/kg body wt, po for 21 days significantly reduced the prostatic weight, theepithelial height and the stromal proliferation in experimental prostatic hypertrophy. Conclusion: Prostane is effectivein the treatment of experimental prostatic hypertrophy in rats and may be passed on to clinical trials on benign prostatichypertrophy after necessary toxicological evaluations. (Asian J Androl 1999 Dec; 1: 175-179)
文摘Objective Localization of the glutathione dependent Nitroblue tetrazolium (NBT) reductase in fresh frozen sections of mouse skin and possible dependence of NBT reductase on tissue thiol levels has been investigated. Methods The fresh frozen tissue sections (8m thickness) were prepared and incubated in medium containing NBT, reduced glutathione (GSH) and phosphate buffer. The staining for GSH was performed with mercury orange. Results The activity of the NBT-reductase in mouse skin has been found to be localized in the areas rich in glutathione and actively proliferating area of the skin. Conclusion The activity of the NBT-reductase seems to be dependent on the glutathione contents.
基金Supported by A grant from Foundation of High Level Talented Specialists of Guizhou Province,China,No. TZJF-200850a grant from Foundation of the Program for Tackling Key Problems of Guizhou Science and Technology Department,China,No. 2010GZ97666
文摘AIM:To investigate the effects of blueberry on hepatic fibrosis and NF-E2-related factor 2(Nrf2) transcription factor in rats.METHODS:Forty-five male Sprague-Dawley rats were randomly divided into control group(A);CCl4-induced hepatic fibrosis group(B);blueberry prevention group(C);Dan-shao-hua-xian capsule(DSHX) prevention group(D);and blueberry + DSHX prevention group(E).Liver fibrosis was induced in rats by subcutaneous injection of CCl4 and a high-lipid/low-protein diet for 8 wk(except the control group).The level of hyaluronic acid(HA) and alanine aminotransferase(ALT) in serum was examined.The activity of superoxide dismutase(SOD),glutathione-S-transferase(GST) and malondialdehyde(MDA) in liver homogenates was determined.The degree of hepatic fibrosis was evaluated by hematoxylin and eosin and Masson staining.Expression of Nrf2 and NADPH quinone oxidoreductase 1(Nqo1) was detected by real-time reversed transcribed-polymerase chain reaction,immunohistochemical techniques,and western blotting.RESULTS:Compared with group B,liver indices,levels of serum HA and ALT of groups C,D and E were reduced(liver indices:0.038 ± 0.008,0.036 ± 0.007,0.036 ± 0.005 vs 0.054 ± 0.009,P<0.05;HA:502.33 ± 110.57 ng/mL,524.25 ± 255.42 ng/mL,499.25 ± 198.10 ng/mL vs 828.50 ± 237.83 ng/mL,P<0.05;ALT:149.44 ± 16.51 U/L,136.88 ± 10.07 U/L,127.38 ± 11.03 U/L vs 203.25 ± 31.62 U/L,P<0.05),and SOD level was significantly higher,but MDA level was lower,in liver homogenates(SOD:1.36 ± 0.09 U/mg,1.42 ± 0.13 U/mg,1.50 ± 0.15 U/mg vs 1.08 ± 0.19 U/mg,P<0.05;MDA:0.294 ± 0.026 nmol/mg,0.285 ± 0.025 nmol/mg,0.284 ± 0.028 nmol/mg vs 0.335 ± 0.056 nmol/mg,P<0.05).Meanwhile,the stage of hepatic fibrosis was significantly weakened(P<0.05).Compared with group A,the activity of GST liver homogenates and expression levels of Nrf2 and Nqo1 in group B were elevated(P<0.05).The expression level of Nrf2 and Nqo1 in groups C,D,and E were increased as compared with group B,but the difference was not significant.CONCLUSION:Blueberry has preventive and protective effects on CCl4-induced hepatic fibrosis by reducing hepatocyte injury and lipid peroxidation.However,these effects may not be related to the activation of Nrf2 during long-term of CCl4.
文摘No study has been performed on identifying microRNAs(miRNAs) and their targets in cotton although cotton is one of the most important fiber and economic crops around the world.In this study,
基金supported by the earmarked fund for the China Agriculture Research System(beef)(CARS-37)the Special Fund for Innovation Team of Modern Agricultural Industrial Technology System in Shandong Province,China(SDAIT-09-09)the Funds of Shandong“Double Tops”Program,China(SYL2017XTTD12)
文摘In order to improve beef color and color stability, step-chilling (SC) was applied on excised bovine Iongissimus lumborum muscle, with chilling starting at 0-4℃ for 5 h, then holding the temperature at 12-18℃ for 6 h, followed by 0-4℃ again until 24 h post-mortem, pH and temperature were measured during rigor on SC loins as well as those subjected to routine chilling (RC, 0-4℃, till 24 h post-mortem). Color L*, a*, b* values, metmyoglobin (MetMb) content, MetMb reducing ability (MRA) and NADH content were determined on samples aged for 1, 7, and 14 d. Sarcoplasmic proteome analysis was only conducted on d 1 samples. The results showed muscles subjected to SC maintained a temperature at around 15℃ for 5 to 10 h post-mortem, and exhibited a slow temperature decline, but rapid pH decline. Beef steaks treated with SC had higher L*, a*, b* and chroma values than those of RC samples at 1 and 7 d chilled storage (0-4℃), while showing no significant difference for a*, b* and chroma values at d 14. The SC samples also exhibited a lower relative content of surface MetMb, higher MRA and NADH content, compared with RC beef steaks during storage, indicating the SC-treated beef showed an improved color stability. Eleven differential protein spots/nine proteins were identified by two-dimensional gel electrophoresis and mass spectrometry, and those proteins were mainly involved in redox, chaperone binding, metabolic and peroxidase activity. Oxidoreductases play a role in decreasing the oxidation-induced myoglobin oxidation and benefiting the production of NADH, and finally improving the colour of beef. Of these, pyruvate dehydrogenase E1 component subunit beta showed a positive correlation with color L*, a*, b* values and accounted for more than 60% of the variation in color values; this protein can be considered as a potential beef color biomarker. The present study provided valuable information for studies on the molecular mechanism of color improvement from step-chilling, as well as for identifying markers associated with beef color.
基金supported by National Natural Science Foundation of China (81272930, 81322038, 31260208, and U1132605)the Science and Technological Key Project of Yunnan Province (2012FB185)West Light Foundation of the Chinese Academy of Sciences (to R.L.)
文摘The WW domain-containing oxidoreductase(WWOX) is a tumor suppressor in a variety of cancers, including breast cancer. Reduced WWOX expression is associated with the basal-like subtype and a relatively poor disease-free survival rate among breast cancer patients. Though several WWOX partners have been identified, the functional mechanisms of WWOX's role in cancers have not been fully addressed to date. In the current study, we found WWOX suppresses expression of KLF5—an oncogenic transcription factor—at protein level, and suppresses cancer cell proliferation in both bladder and breast cancer cell lines. Furthermore, we demonstrated that WWOX physically interacts with KLF5 via the former's WW domains and the latter's PY motifs. Interestingly, we found the expression of WWOX negatively correlates with KLF5 expression in a panel of breast cancer cell lines. Taken together, we conjecture that WWOX may suppress cancer cell proliferation partially by reducing the expression of KLF5.
基金supported by the Autonomous region Natural Science Foundation of China,No. 200711020906
文摘Objective: To investigate the relationship between NAD(P)H:quinone oxidoreductase 1 (NQOI) C609T polymorphism and colon cancer risk in farmers from western region of Inner Mongolia. Methods: Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was performed to analyze NQO1 C609T polymorphism from 160 healthy controls and 76 colon cancer patients. Results: Among the colon cancer patients, the incidence of NQOI T allele (53.29%) was significantly higher than it in control group (33.75%, P〈0.001). The individuals with NQO1 T allele had higher risk [2.239 (95% CI: 1.510-3.321) times] to develop colon cancer than individuals with NQO1 C allele. The incidence of NQO1 (TFI-) (34.21%) in colon cancer patients was higher than that in control group (15.62%, P〈0.001). Odds ratios (OR) analysis suggested that NQO1 (T/F) and NQOI (T/C) genotype carriers had 3.813 (95% CI: 1.836-7.920) times and 2.080 (1.026-4.219) times risk compared with wild-type NQO1 (C/C) gene carriers in developing colon cancer. Individuals with NQO1 (T/I') genotype had 2.541 (95% CI: 0.990-6.552) times, 3.713 (95% CI: 1.542-8.935) times, and 3.471 (95% CI: 1.356-8.886) times risk than individuals with NQOI (T/C) or NQOI (C/C) genotype in well- differentiated, moderately-differentiated, and poorly-differentiated colon cancer patients, respectively. Conclusions: NQO1 gene C609T could be one of risk factors of colon cancer in farmers from western region of Inner Mongolia,
基金supported by the grants from the National Natural Science Foundations of China(Grant Nos.30870192 and 30971744)the National Major Science and Technology Project to Create New Crop Varieties Using Gene Transfer Technology,China(Grant No.2009ZX08001-029B)the Shandong Natural Science Funds for Distinguished Young Scholar,China(Grant No.JQ200911)
文摘Phytochromes in rice are encoded by a gene family composed of three members, PHYA, PHYB, and PHYC. Through characterizing the phytochrome mutants and wild type (WT) in terms of photomorphogenesis, roles of individual phytochromes have been preliminarily explored in regulating rice de-etiolation, flowering time and fertility. However, little information has been reported about whether or how phytochromes affect chlorophyll biosynthesis and chloroplast development in rice. In this study, we compared the chlorophyll contents of wild type and the phyA, phyB and phyAphyB mutants grown under either white light (WL) or red light (R). The results suggest that phyB perceives R to positively regulate chlorophyll biosynthesis, while the role of phyA can be detected only in the phyB-deficient mutant. Analyses of the expression levels of genes involved in chlorophyll biosynthesis revealed that phytochromes affected the chlorophyll biosynthesis by regulating protochlorophyll oxidoreductase A (PORA) expression. The role of phyB in chloroplast development was also analyzed, and the results suggest that phyB perceives R to regulate chloroplast development by affecting the numbers of chloroplasts and grana, as well as the chloroplast membrane system.
基金China Ocean Mineral Resources R&D Association under contract No. DYXM-115-02-2-19Basic Scientific Fund for National Public Research Institutes of China under contract No. 2011T04
文摘CO2 fixation pathway of Caminibacter profundus, a chemolithoautotrophic e-Proteobacteria from deep-sea hydrothermal vent, was determined and characterized by genetic and enzymatic analyses. Gene expression of key enzymes for CO2 fixation in response to salinity, pH and O2 in Medium 829 were also investigated. The results demonstrate that C. profundus contained aclB, porA and oorA, the genes encoding key enzymes of reductive tricarboxylic acid (rTCA) cycle. However, genes fragments of cbbL and cbbMencoding key enzyme of Calvin cycle were not recovered. Key enzymatic activities of ATP citrate lyase (ACL), pyruvate: ferredoxin oxidoreductase (POR) and 2-oxoglutarate: ferredoxin oxidoreductase (OOR) were also present in C. profun- dus. The combination of genetic and enzymatic analyses confirm that C. profundus adopted rTCA cycle for carbon assimilation. The results of aclB and oorA relative expressions of C. profundus demonstrate that the ranges of environmental factors for high genes expression were sea salt 3.0%-5.0% (optimum 3.0%), pH 5.0-6.5(optimum pH 6.5), anaerobic to microaerobic conditions (optimum 1.0% 02). Gene expression pat- terns under different conditions show similar patterns with bacterial growth, revealing that key rTCA cycle genes provided molecular basis for bacterial growth and propagation. Our results suggest that C. profun- dus could regulate key genes of rTCA cycle for carbon assimilation and energy metabolism in response to environmental fluctuations in hydrothermal vent.
基金grants from the National Natural Science Foundation of China(No.81560222)the Guizhou Science and Technology Foundation(No.[2017]7187,and No.[2013]2043).
文摘The present study aimed to explore the molecular mechanisms underlying the increase of nicotinamide adenine dinucleotide phosphate:quinine oxidoreductase 1(NQO1)and y-glutamylcysteine synthetase(γ-GCS)in brain tissues after intracerebral hemorrhage(ICH).The microglial cells obtained from newborn rats were cultured and then randomly divided into the normal control group(NC group),model control group(MC group),rosiglitazone(RSG)intervention group(RSG group),retinoic-acid intervention group(RSG+RA group),and sulfbraphane group(RSG+SF group).The expression levels of NQO1,γ-GCS,and nuclear factor E2-related factor 2(Nrf2)were measured by real-time polymerase chain reaction(RT-PCR)and Western blotting,respectively.The results showed that the levels of NQO1,γ-GCS and Nrf2 were significantly increased in the MC group and the RSG group as compared with those in the NC group(P<0.01).They were found to be markedly decreased in the RSG+RA group and increased in the RSG+SF group when compared with those in the MC group or the RSG group(P<0.01).The RSG+SF group displayed the highest levels of NQO1,γ-GCS,and Nrf2 among the five groups.In conclusion,a medium dose of RSG increased the anti-oxidative ability of thrombinactivated microglia by increasing the expression of NQO1 and γ-GCS.The molecular mechanisms underlying the increase of NQO1 and γ-GCS in thrombin-activated microglia may be associated with the activation of Nrf2.
基金Supported by the National Natural Science Foundation of China,No.31971188 and No.81773189the Nature Science Foundation of Zhejiang Province,China,No.LQ16H160004 and No.LY17H270002The Hygiene Department of Zhejiang,No.2016KYB139.
文摘BACKGROUND Quinine oxidoreductase 1(NQO1)plays a vital role in protecting normal cells against oxidative damage and electrophilic attack.It is highly expressed in many solid tumors,suggesting a role in cancer development and progression.However,the role of NQO1 in gastric cancer and its effect on cancer development and prognosis have not been fully investigated.AIM To investigate the clinical relevance of NQO1 protein expression in gastric cancer and to explore the potential of NQO1 to serve as a prognostic biomarker and therapeutic target.METHODS In this retrospective study,gastric cancer specimens of 175 patients who were treated between 1995 and 2011 were subjected to immunohistochemistry analyses for NQO1.The correlation of NQO1 expression with gastric cancer prognosis and clinical and pathological parameters was investigated.RESULTS NQO1 protein was overexpressed in 59.43%(104/175)of the analyzed samples.Overexpression of NQO1 was associated with a significantly inferior prognosis.In addition,multivariate analysis suggested that NQO1 overexpression,along with tumor stage and patient age,are prominent prognostic biomarkers for gastric cancer.Moreover,NQO1 overexpression was correlated to a better response to 5-fluorouracil(5-FU)-based adjuvant chemotherapy.CONCLUSION NQO1 overexpression is associated with a significantly poor prognosis and better response to 5-FU in patients with gastric cancer.These findings are relevant for improving therapeutic approaches for gastric cancer patients.
文摘The growth-inhibiting and apoptosis-inducing effects of WW domain-containing oxidoreductase(WWOX) gene on ovarian cancer cell line A2780 were investigated.The full length cDNA of human WWOX gene was amplified from normal human ovary tissues.The correct cDNA of full length WWOX was subcloned into eukaryocytic expression vector pCMV.After introduction of WWOX gene into cancer cells with liposome,the WWOX mRNA and protein level in the cancer cells were detected by reverse transcription polymerase chain reaction(RT-PCR) and immunoblotting.The growth activities of cancer cells were detected by Trypan blue staining.The clone formation assay in soft agar was employed to observe the proliferation of the cancer cells.Apoptosis was examined by DNA ladder and acridine orange-ethidium bromide fluorescent staining.The results showed that 72 h after WWOX gene transfection,the WWOX expression was increased significantly(P<0.01).The growth of ovarian cancer cells was decreased by 16.41% to 38.49%(P<0.01).The clone formation abilities were reduced(P<0.01).Some cancer cells presented the characteristic morphological changes of apoptosis with obvious ladder bands on electrophoresis.The apoptosis rate was(20.7±6.0)%(P<0.01).It was concluded that over-expression of WWOX gene could induce apoptosis and inhibit the growth of ovarian cancer cells,which might be potentially useful in the gene therapy of ovarian cancers.