Aflatoxin B_1(AFB_1)is a common contaminant in cereals of global concern,and long-term low-dose exposure can adversely affect human health.Here,we showed that populations with dietary patterns characterized by high-fa...Aflatoxin B_1(AFB_1)is a common contaminant in cereals of global concern,and long-term low-dose exposure can adversely affect human health.Here,we showed that populations with dietary patterns characterized by high-fat diet(HFD)might have an increased risk of exposure to high levels of AFB_1.Our data indicated that chronic exposure of AFB_1 induced“gut-liver axis”injury in mice under HFD and normal diet(ND)patterns.AFB_1 further aggravated hepatic and intestinal injury,and intestinal microbiota disruption in HFD mice.Bifidobacterium breve BAA-2849 intervention analysis showed that liver injury and lipid disorders caused by AFB_1 exposure were alleviated by regulating the proportions of different gut microbes.We demonstrated through a mice model that the populations with a dietary pattern of HFD might be more susceptible to AFB_1 exposure and adverse effects on the gut-liver axis,and the toxicity of AFB_1 exposure can be alleviated by regulating the gut microbiota.展开更多
AIM:To determine global DNA methylation in paired hepatocellular carcinoma(HCC) samples using several different assays and explore the correlations between hypomethylation and clinical parameters and biomarkers,includ...AIM:To determine global DNA methylation in paired hepatocellular carcinoma(HCC) samples using several different assays and explore the correlations between hypomethylation and clinical parameters and biomarkers,including that of aflatoxin B 1 exposure.METHODS:Using the radio labeled methyl acceptance assay as a measure of global hypomethylation,as well as two repetitive elements,including satellite 2(Sat2) by MethyLight and long interspersed nucleotide elements(LINE1),by pyrosequencing.RESULTS:By all three assays,mean methylation levels in tumor tissues were significantly lower than that in adjacent tissues.Methyl acceptance assay log(mean ± SD) disintegrations/min/ng DNA are 70.0 ± 54.8 and 32.4 ± 15.6,respectively,P = 0.040;percent methylation of Sat2 42.2 ± 55.1 and 117.9 ± 88.8,respectively,P < 0.0001 and percent methylation LINE1 48.6 ± 14.8 and 71.7 ± 1.4,respectively,P < 0.0001.Aflatoxin B 1 albumin(AFB 1-Alb) adducts,a measure of exposure to this dietary carcinogen,were inversely correlated with LINE1 methylation(r =-0.36,P = 0.034).CONCLUSION:Consistent hypomethylation in tumor compared to adjacent tissue was found by the three different methods.AFB 1 exposure is associated with DNA global hypomethylation,suggesting that chemical carcinogens may influence epigenetic changes in humans.展开更多
Some unique subclasses of Camelidae antibodies are devoid of the light chain, and the antigen binding site is comprised exclusively of the variable domain of the heavy chain （VHH）. The recombinant VHHs have a high p...Some unique subclasses of Camelidae antibodies are devoid of the light chain, and the antigen binding site is comprised exclusively of the variable domain of the heavy chain （VHH）. The recombinant VHHs have a high potential as alternative reagents for the next generation of immunoassay. In particular, they might be very useful for molecular mimicry. The present study demonstrated an alpaca immunized with the F（ab＇）z fragment of anti-aflatoxin B1 mAb and developed an important anti-idiotypic （anti-ld） responses. Antigen-specific elution method was used for panning private anti-ld VHHs from the constructed alpaca VHH library. The selected VHHs were expressed, renatured, purified, and then identified by a competitive enzyme-linked immunosorbent assay （ELISA）. Our findings indicated that the VHH would be an alternative tool for haptens mimicry studies.展开更多
Objective： The aim of this study was to study the effect of Ginkgo biloba extract （EGb761） on metabolism of afiatoxin B1 （AFB1） in Wistar rats. Methods： Seventy one Wistar rats were assigned at random to groups ...Objective： The aim of this study was to study the effect of Ginkgo biloba extract （EGb761） on metabolism of afiatoxin B1 （AFB1） in Wistar rats. Methods： Seventy one Wistar rats were assigned at random to groups A, B and C. Rats in groups A, B were injected with AFB1 （intraperitoneal, 100-200 ug/kg body weight, 1-3 times/week）. Group C was normal control. Rats in group B were fed in food with EGb761, while rats in groups A, C were given normal food. Blood samples were collected and liver biopsies were performed on the 14th, 28th and 42nd week. All the rats were sacrificed on the 64th week. The incidence of hepatocarcinoma was investigated. The hepatic phase I drug-metabolizing enzyme Cytochrome-P450 （CYP450） and phase II metabolizing enzyme glutathione S-transferase （GST） were analyzed with spectrometry. Serum AFB1- lysine adduct levels were assessed with high performance liquid chromatography （HPLC）. The expression of 8-hydroxydeoxy- guanosine （8-OHdG） was measured with immunohistochemistry. Results： The incidence of hepatocellular carcinoma （HCC） in group B was significantly lower than that in group A （26.92% vs 76.00%, P 〈 0.001）. No HCC developed in group C. EGb761 showed no effects on the activities of CYP450 and GST in rat liver tissues. The level of AFB1-lysine adduct reached the peak （4356.01 pg/mg albumin） at the 14th week in group A. EGb761 significantly inhibited the formation of AFB1-lysine adduct in serum by 13.07% at the 14th week （P = 0.033）, and 73.63% at the 42nd week （P = 0.002）. The expression of 8-OHdG protein in rat liver tissues in group B was significantly lower than that in group A at the 28th, 42nd, and 64th week （P 〈 0.05）. Conclusion： The main mechanism underlying the effect of EGb761 in blocking hepatocarcinogenesis induced by AFB1 may not be fully attributable to its influence on the activity of liver phase I and phase II metabolizing enzymes. EGb761 inhibits the production of AFB1-lysine adducts, decreases the expression of 8-OHdG protein, and finally alleviates the DNA oxidative injury, which may be one of the mechanisms for the effects of EGb761 in inhibiting or delaying AFB1-induced hepatocarcinogenesis.展开更多
In this study, the extraction process of aflatoxin B~ from tartary buckwheat bran was optimized with ELISA detection method to determine the optimal conditions for extracting aflatoxin B1 from tartary buckwheat bran. ...In this study, the extraction process of aflatoxin B~ from tartary buckwheat bran was optimized with ELISA detection method to determine the optimal conditions for extracting aflatoxin B1 from tartary buckwheat bran. The results of standard recovery test of blank solvent and sample confirmed the feasibility of ELISA detection method. Orthogonal experiment was performed to optimize the solid-liquid ratio, ultrasonic extraction time and ultrasonic amplitude. The results show that it is feasible to detect aflatoxin B1 content with ELISA method. The optimal ultrasonic extraction conditions were ： methanol-water ratio 6： 4, solld-liquid ratio 1 g： 5 ml, ultrasonic extraction time 15 min, ultrasonic amplitude 15 ~. Under the optimized conditions, 1 065.1 ng/L aflatoxin B1 was extracted from tartary buckwheat bran.展开更多
Laboratory animals maintained on a reduced calorie but nutritionally adequate diet have extended life spans and lowered incidences of spontaneous and chemically induced cancers compared to ad libitum- fed counterparts...Laboratory animals maintained on a reduced calorie but nutritionally adequate diet have extended life spans and lowered incidences of spontaneous and chemically induced cancers compared to ad libitum- fed counterparts. Many of the effects of dietary restriction on laboratory animals have been suggested to be related to a deceleration of the aging process. The inhibition of age-related changes in xenobiotic metabolizing enzyme activities by dietary restriction has previously been reported. Alterations of these enzyme activities may cause changes in metabolic activation of carcinogens and, therefore, carcinogen-DNA binding. DNA-repair capability has also been reported to be enhanced in diet-restricted rats. Using AFB1 as a model carcinogen, we have studied in vivo and in vitro hepatic AFB1 -DNA binding, demonstrating that dietary restriction (60% of ad libitum consumption) may decrease the metabolic activation of AFB1, and subsequently reduce AFB 1-DNA binding. Our preliminary results obtained from the AFB 1-DNA binding experiments in isolated hepatocytes suggest that the observed age-dependent reduction in AFB 1-DNA binding which may be attributed to a loss of metabolic activating capability was delayed in the diet-restricted rats.展开更多
Background This study was carried out to investigate the individual and combined contamination of aflatoxin B_(1)(AFB_(1)),deoxynivalenol(DON),and zearalenone(ZEN)in feeds in China between 2021 and 2024.A total of 23,...Background This study was carried out to investigate the individual and combined contamination of aflatoxin B_(1)(AFB_(1)),deoxynivalenol(DON),and zearalenone(ZEN)in feeds in China between 2021 and 2024.A total of 23,003 feed samples,including 17,489 feedstuff samples and 5,514 complete feed samples,were collected from different provinces of China for mycotoxin analysis.Results The analyzed mycotoxins displayed considerably high contamination in the feed samples,with the individual contamination of AFB_(1),DON,and ZEN were 20.0%–100%,33.3%–100%,and 85.0%–100%,respectively.The average concentrations of AFB_(1),DON,and ZEN were 1.2–728.7μg/kg,106–8,634.8μg/kg,and 18.1–3,341.6μg/kg,respectively.Notably,the rates over China’s safety standards for AFB_(1),DON,and ZEN in raw ingredients were 9.7%,2.7%,and 15.7%,respectively.Meanwhile,3.5%,1.1%,and 8.7%of analyzed complete feeds exceeded China’s safety standards for AFB_(1),DON,and ZEN,respectively.Moreover,the co-contamination rates of AFB_(1),DON,and ZEN in more than 70%of raw ingredients and 87.5%of complete feed products were 60.0%–100%and 61.5%–100%,respectively.Conclusion This study reveals that the feeds in China have commonly been contaminated with AFB_(1),DON,and ZEN alone and their combination during the past four years.These findings highlight the significance of monitoring mycotoxin contaminant levels in domestic animal feed and the importance of carrying out feed administration and remediation strategies for mycotoxin control.展开更多
Background AFB_(1)-8,9-exo-epoxide(AFBO)is the highly toxic product of Aflatoxin B_(1)(AFB_(1)).Glutathione S-transferases(GSTs)play pivotal roles in detoxifying AFB_(1) by catalyzing the conjugation of AFBO with glut...Background AFB_(1)-8,9-exo-epoxide(AFBO)is the highly toxic product of Aflatoxin B_(1)(AFB_(1)).Glutathione S-transferases(GSTs)play pivotal roles in detoxifying AFB_(1) by catalyzing the conjugation of AFBO with glutathione(GSH).Although there are over 20 GST isozymes that have been identified in chicken,GST isozymes involved in the detoxification process of AFB_(1) have not been identified yet.The objective of this study was to determine which GST isozymes played key role in detoxification of AFB_(1).Results A total of 17 pcDNA3.1(+)-GST isozyme plasmids were constructed and the GST isozyme genes were overexpressed by 80–2,500,000 folds in the chicken Leghorn male hepatoma(LMH)cells.Compared to the AFB_(1) treatment,overexpression of GSTA2X,GSTA3,GSTT1L,GSTZ1-1,and GSTZ1-2 increased the cell viability by 6.5%–17.0%in LMH cells.Moreover,overexpression of five GST isozymes reduced the release of lactate dehydrogenase and reactive oxygen species by 8.8%–64.4%,and 57.2%–77.6%,respectively,as well as enhanced the production AFBO-GSH by 15.8%–19.6%,thus mitigating DNA damage induced by AFB_(1).After comprehensive evaluation of various indicators,GSTA2X displayed the best detoxification effects against AFB_(1).GSTA2X was expressed in Pichia pastoris X-33 and its enzymatic properties for catalyzing the conjugation of AFBO with GSH showed that the optimum temperature and pH were 20–25℃ and 7.6–8.6 as well as the enzymatic kinetic parameter V_(max) was 0.23 nmol/min/mg and the Michaelis constant was 86.05μmol/L with the AFB_(1) as substrate.Conclusions In conclusion,GSTA2X,GSTA3,GSTT1L,GSTZ1-1,and GSTZ1-2 played key roles in AFB_(1) detoxification,which will provide new remediation strategies to prevent aflatoxicosis in chickens.展开更多
Background Aflatoxin B_(1)(AFB_(1))risks animal and human health,and the liver is considered the most crucial detoxification organ.Phlorotannin(PT)is a polyhydroxy phenol that has a wide range of biological activities...Background Aflatoxin B_(1)(AFB_(1))risks animal and human health,and the liver is considered the most crucial detoxification organ.Phlorotannin(PT)is a polyhydroxy phenol that has a wide range of biological activities,including antioxidation and hepatoprotection,which can promote the ability of liver detoxification.This study aimed to elucidate the protective effect of PT on AFB_(1)-induced liver damage in broilers.Results In vivo experiment showed that the PT reduced AFB_(1) content and AFB_(1)-exo-8,9-epoxide DNA(AFBODNA)concentration in serum and liver(P<0.05),improved the histomorphology of liver and hepatic mitochondria,and activated nuclear factor erythroid 2-related factor 2(Nrf2)-related antioxidant and detoxification pathway by upregulating the activities of antioxidant enzymes(catalase[CAT],glutathione S-transferase[GST])and total antioxidant capacity(T-AOC)level(P<0.05),and inhibited the mRNA expression of CYP1A1(cytochrome P450 family 1 subfamily A member 1)and phase Ⅱ detoxification enzyme related genes(GPX1,GSTT1,and NQO1)of broilers exposed to AFB_(1)(P<0.05).Meanwhile,PT upregulated the Nrf1 pathway-related mitochondrial biosynthetic genes(Nrf1,mitochondrial transcription factor A[TFAM],mitofusin 1[MFN1])in broilers fed AFB_(1) contaminated diet(P<0.05).In vitro verification study suggested that the use of Nrf2/Nrf1 inhibitors suppressed the ameliorative role of PT on AFB_(1)-induced liver injury of broilers,which was manifested in the mRNA expression of Nrf2,NQO1,GSTT3,Nrf1,TFAM,and other genes decreasing(P<0.05),and down-regulation of the protein expression of Nrf2,total and nucleus p-Nrf2,and total and nucleus p-Nrf1(P<0.05).Conclusion The PT ameliorates oxidative stress and hepatotoxicity by activating the Nrf2-mediated phase Ⅱ detoxification enzymes pathway and maintains mitochondrial homeostasis by activating the Nrf1 signaling pathway in broilers exposed to AFB_(1).展开更多
Aflatoxin B1(AFB1)is a carcinogenic toxin naturally produced in most food crops that severely threaten human health,and effective methods are urgent to improve the detection accuracy.Herein an indirect competitive imm...Aflatoxin B1(AFB1)is a carcinogenic toxin naturally produced in most food crops that severely threaten human health,and effective methods are urgent to improve the detection accuracy.Herein an indirect competitive immunosorbent approach was elaborately developed based on high-affinity immunoglobulin G(IgG)coupled CuO-anchored Fe_(3)O_(4)nanozymes for precise and ultrasensitive detection of AFB_(1)in food crops including peanut,corn and wheat.The high-affinity nanozymes were fabricated by the assembly of inner core Fe_(3)O_(4)nanoparticles and mesoporous silica capping layer,Cu O further situated within large aperture of the coating layer via in-situ growth,and then conjugated with ligand rabbit anti-mouse Ig G,which can specifically bind with AFB_(1).The results showed the hybrid high-affinity nanozymes displayed enhanced peroxidasemimic activities and catalytic performances,achieving a linear range of 0.06-61.93(lg(ng/mL))and a detection limit of 0.0037 ng/mL,30 times better than that of the conventional enzyme-linked immunosorbent assay.The constructed nanozymes were successfully applied to the detection of AFB_(1)in food products with an average spiked recovery of 96.53%and relative standard deviations less than 2.8%.Therefore,the accurate hybrid nanozymes may serve for AFB_(1)detection in various foods in future.展开更多
Background Aflatoxins have been reported as a significant pollutant in feed,capable of causing harm to the liver,gastrointestinal tract and kidneys of piglets.However,research on the interactions among aflatoxin B1(AF...Background Aflatoxins have been reported as a significant pollutant in feed,capable of causing harm to the liver,gastrointestinal tract and kidneys of piglets.However,research on the interactions among aflatoxin B1(AFB1),bile acid(BA)metabolism and gut microbiota is limited.Methods In this study,piglets were treated with AFB1 and antibiotics(ABX)to evaluate the interaction between AFB1 and gut microbiota.Subsequently,the roles of the farnesoid X receptor(FXR)and sterol 12α-hydroxylase(CYP8B1)in AFB1 absorption were studied by using FXR agonists obeticholic acid(OCA)and Cyp8b1-knockout(KO)mice,respectively.Result AFB1 inhibited bile salt hydrolase(BSH)activity in ileal microbiota,downregulated ileal FXR expression,and upregulated CYP8B1 expression in liver,increasing the proportion of 12α-OH BAs and potentially enhancing AFB1 absorption.ABX treatment reduced AFB1 absorption and liver damage,and unexpectedly increased BSH activity,counteracting the AFB1-induced downregulation of FXR and upregulation of CYP8B1.OCA reactivated ileal FXR,reduced AFB1 absorption,and alleviated liver damage.Furthermore,Cyp8b1-KO mice showed increased resistance to AFB1-induced liver damage by lowering AFB1 absorption.Conclusions These results underscore the significance of gut microbiota and BAs in AFB1 absorption,suggesting new strategies to mitigate health risks from AFB1 in piglets.展开更多
Hepatocellular carcinoma(HCC)is a major global health challenge,particularly in regions with high aflatoxin B1(AFB1)exposure.This editorial explores the mechanistic interplay between AFB1 and tissue inhibitor of metal...Hepatocellular carcinoma(HCC)is a major global health challenge,particularly in regions with high aflatoxin B1(AFB1)exposure.This editorial explores the mechanistic interplay between AFB1 and tissue inhibitor of metalloproteinase-3(TIMP-3)in AFB1-related HCC.TIMP-3,frequently downregulated in HCC due to promoter methylation,is linked to increased tumor aggressiveness and poor prognosis.We propose that AFB1 induces epigenetic silencing of TIMP-3,potentially via DNA adducts and oxidative stress,exacerbating AFB1-related HCC progression.This AFB1-TIMP-3 axis highlights TIMP-3’s potential as a prognostic biomarker and therapeutic target.Future research should focus on elucidating these molecular pathways and integrating TIMP-3 into clinical practice for early detection and targeted therapies in AFB1-prevalent regions.展开更多
Aflatoxins B1(AFB1)contamination in agro-food holds great threaten to human and animal health.Conventional test strips for rapid AFB1 visualized monitoring remains challenged by improvement of sensitivity and matrix i...Aflatoxins B1(AFB1)contamination in agro-food holds great threaten to human and animal health.Conventional test strips for rapid AFB1 visualized monitoring remains challenged by improvement of sensitivity and matrix interference resistance.In this case,we developed a portable electrochemiluminescence(ECL)imaging test strip with dual-signal outputs for AFB1 quantification in corn samples.RuPEI@SiO_(2)@Au nanospheres were synthesized for bonding with anti-AFB1 antibody and then colorimetrical signal-reported on test line through the capillary flow at strips.Meanwhile,ECL imaging signal of the constructed carbon-ink-based working electrode on polyvinyl chloride substrate of strips was exported under an applied potential of 1.25 V.The whole ECL test strips not only endowed convenient colorimetric responses but guaranteed quick-witted ECL image distinguishment even at extremely low AFB1 content.The detection limit of this ECL imaging-integrated mode was 10-fold lower than that of only colorimetric mode.Furthermore,satisfactory selectivity,reliability and practicability of the as-proposed ECL test strips were demonstrated.This work offered a promising platform for on-site,accurate and sensitive detection of pollutants in foods.展开更多
Given severe health-hazardous effects of aflatoxin B1(AFB1) widely occurring in cereal grains and animal feeds,it is highly urgent to develop analytical methods for its rapid screening.In this work,we proposed a simpl...Given severe health-hazardous effects of aflatoxin B1(AFB1) widely occurring in cereal grains and animal feeds,it is highly urgent to develop analytical methods for its rapid screening.In this work,we proposed a simple and high-throughput method for the determination of AFB1 in millet and buckwheat samples using high performance thin layer chromatography(HPTLC) linked to fluorescence densitometry.The first step was to optimize the solid-liquid extraction for the crude clean-up of the samples.The QuEChERS(Quick,Easy,Cheap,Effective,Robust and Safe) extraction strategy was used and different solvent systems for their extraction efficiency of AFB1 from the samples were evaluated.Then,trichloromethane:ethyl acetate(7:3,V/V) was used as the mobile phase to realize the separation of the targeted compound from background noises on silica gel plates.Quantification was readily performed with densitometry in fluorescence mode.In order to fix the optimal excitation wavelength,spectra scanning ranging 250-400 nm was carried out,revealing that 364 nm light gave the highest signal.With the optimized optical system,high sensitivity to AFB1 was achieved,with a limit of detection(LOD) at 3 μg/kg.Apart from that,good linearity(0.999) was obtained within the range of 1-80 ng/band of AFB 1.To assess the analysis accuracy,2 levels of AFB 1(50 and100 μg/kg) were spiked into real grain samples.The obtained results showed that the recovery rates were within the range of 81.6%-114.0%.The proof-of-concept results of this work evidenced that HPTLC is a promising analytical tool for the screening of mycotoxin in difficult samples.展开更多
基金supported by grants from the National Natural Science Foundation of China(32125031)the Fundamental Research Funds for the Central Universities(JUSRP222001)Collaborative Innovation Center for Food Safety and Quality Control,China。
文摘Aflatoxin B_1(AFB_1)is a common contaminant in cereals of global concern,and long-term low-dose exposure can adversely affect human health.Here,we showed that populations with dietary patterns characterized by high-fat diet(HFD)might have an increased risk of exposure to high levels of AFB_1.Our data indicated that chronic exposure of AFB_1 induced“gut-liver axis”injury in mice under HFD and normal diet(ND)patterns.AFB_1 further aggravated hepatic and intestinal injury,and intestinal microbiota disruption in HFD mice.Bifidobacterium breve BAA-2849 intervention analysis showed that liver injury and lipid disorders caused by AFB_1 exposure were alleviated by regulating the proportions of different gut microbes.We demonstrated through a mice model that the populations with a dietary pattern of HFD might be more susceptible to AFB_1 exposure and adverse effects on the gut-liver axis,and the toxicity of AFB_1 exposure can be alleviated by regulating the gut microbiota.
基金Supported by A grant from the National Institute of Health,No. ES005116 and No.P30ES009089
文摘AIM:To determine global DNA methylation in paired hepatocellular carcinoma(HCC) samples using several different assays and explore the correlations between hypomethylation and clinical parameters and biomarkers,including that of aflatoxin B 1 exposure.METHODS:Using the radio labeled methyl acceptance assay as a measure of global hypomethylation,as well as two repetitive elements,including satellite 2(Sat2) by MethyLight and long interspersed nucleotide elements(LINE1),by pyrosequencing.RESULTS:By all three assays,mean methylation levels in tumor tissues were significantly lower than that in adjacent tissues.Methyl acceptance assay log(mean ± SD) disintegrations/min/ng DNA are 70.0 ± 54.8 and 32.4 ± 15.6,respectively,P = 0.040;percent methylation of Sat2 42.2 ± 55.1 and 117.9 ± 88.8,respectively,P < 0.0001 and percent methylation LINE1 48.6 ± 14.8 and 71.7 ± 1.4,respectively,P < 0.0001.Aflatoxin B 1 albumin(AFB 1-Alb) adducts,a measure of exposure to this dietary carcinogen,were inversely correlated with LINE1 methylation(r =-0.36,P = 0.034).CONCLUSION:Consistent hypomethylation in tumor compared to adjacent tissue was found by the three different methods.AFB 1 exposure is associated with DNA global hypomethylation,suggesting that chemical carcinogens may influence epigenetic changes in humans.
基金supported by a grant from the National Basic Research Program of China(2013CB127804)the National Natural Science Funds(31171696,China)the Research Program of the State Key Laboratory of Food Science and Technology,Nanchang University(SKLF-MB-201002)
文摘Some unique subclasses of Camelidae antibodies are devoid of the light chain, and the antigen binding site is comprised exclusively of the variable domain of the heavy chain （VHH）. The recombinant VHHs have a high potential as alternative reagents for the next generation of immunoassay. In particular, they might be very useful for molecular mimicry. The present study demonstrated an alpaca immunized with the F（ab＇）z fragment of anti-aflatoxin B1 mAb and developed an important anti-idiotypic （anti-ld） responses. Antigen-specific elution method was used for panning private anti-ld VHHs from the constructed alpaca VHH library. The selected VHHs were expressed, renatured, purified, and then identified by a competitive enzyme-linked immunosorbent assay （ELISA）. Our findings indicated that the VHH would be an alternative tool for haptens mimicry studies.
文摘Objective： The aim of this study was to study the effect of Ginkgo biloba extract （EGb761） on metabolism of afiatoxin B1 （AFB1） in Wistar rats. Methods： Seventy one Wistar rats were assigned at random to groups A, B and C. Rats in groups A, B were injected with AFB1 （intraperitoneal, 100-200 ug/kg body weight, 1-3 times/week）. Group C was normal control. Rats in group B were fed in food with EGb761, while rats in groups A, C were given normal food. Blood samples were collected and liver biopsies were performed on the 14th, 28th and 42nd week. All the rats were sacrificed on the 64th week. The incidence of hepatocarcinoma was investigated. The hepatic phase I drug-metabolizing enzyme Cytochrome-P450 （CYP450） and phase II metabolizing enzyme glutathione S-transferase （GST） were analyzed with spectrometry. Serum AFB1- lysine adduct levels were assessed with high performance liquid chromatography （HPLC）. The expression of 8-hydroxydeoxy- guanosine （8-OHdG） was measured with immunohistochemistry. Results： The incidence of hepatocellular carcinoma （HCC） in group B was significantly lower than that in group A （26.92% vs 76.00%, P 〈 0.001）. No HCC developed in group C. EGb761 showed no effects on the activities of CYP450 and GST in rat liver tissues. The level of AFB1-lysine adduct reached the peak （4356.01 pg/mg albumin） at the 14th week in group A. EGb761 significantly inhibited the formation of AFB1-lysine adduct in serum by 13.07% at the 14th week （P = 0.033）, and 73.63% at the 42nd week （P = 0.002）. The expression of 8-OHdG protein in rat liver tissues in group B was significantly lower than that in group A at the 28th, 42nd, and 64th week （P 〈 0.05）. Conclusion： The main mechanism underlying the effect of EGb761 in blocking hepatocarcinogenesis induced by AFB1 may not be fully attributable to its influence on the activity of liver phase I and phase II metabolizing enzymes. EGb761 inhibits the production of AFB1-lysine adducts, decreases the expression of 8-OHdG protein, and finally alleviates the DNA oxidative injury, which may be one of the mechanisms for the effects of EGb761 in inhibiting or delaying AFB1-induced hepatocarcinogenesis.
基金Supported by Key Science and Technology Project of Education Department of Sichuan Province(13ZA0157)
文摘In this study, the extraction process of aflatoxin B~ from tartary buckwheat bran was optimized with ELISA detection method to determine the optimal conditions for extracting aflatoxin B1 from tartary buckwheat bran. The results of standard recovery test of blank solvent and sample confirmed the feasibility of ELISA detection method. Orthogonal experiment was performed to optimize the solid-liquid ratio, ultrasonic extraction time and ultrasonic amplitude. The results show that it is feasible to detect aflatoxin B1 content with ELISA method. The optimal ultrasonic extraction conditions were ： methanol-water ratio 6： 4, solld-liquid ratio 1 g： 5 ml, ultrasonic extraction time 15 min, ultrasonic amplitude 15 ~. Under the optimized conditions, 1 065.1 ng/L aflatoxin B1 was extracted from tartary buckwheat bran.
文摘Laboratory animals maintained on a reduced calorie but nutritionally adequate diet have extended life spans and lowered incidences of spontaneous and chemically induced cancers compared to ad libitum- fed counterparts. Many of the effects of dietary restriction on laboratory animals have been suggested to be related to a deceleration of the aging process. The inhibition of age-related changes in xenobiotic metabolizing enzyme activities by dietary restriction has previously been reported. Alterations of these enzyme activities may cause changes in metabolic activation of carcinogens and, therefore, carcinogen-DNA binding. DNA-repair capability has also been reported to be enhanced in diet-restricted rats. Using AFB1 as a model carcinogen, we have studied in vivo and in vitro hepatic AFB1 -DNA binding, demonstrating that dietary restriction (60% of ad libitum consumption) may decrease the metabolic activation of AFB1, and subsequently reduce AFB 1-DNA binding. Our preliminary results obtained from the AFB 1-DNA binding experiments in isolated hepatocytes suggest that the observed age-dependent reduction in AFB 1-DNA binding which may be attributed to a loss of metabolic activating capability was delayed in the diet-restricted rats.
基金supported by the Chinese Natural Science Foundation projects(32272915 and 32472949)National Key Research and Development Programs of China(2023YFD1301003)+1 种基金the Fundamental Research Funds for the Central Universities(2662023DKPY002)Hebei Panshuo Biotechnolog Co.,Ltd.
文摘Background This study was carried out to investigate the individual and combined contamination of aflatoxin B_(1)(AFB_(1)),deoxynivalenol(DON),and zearalenone(ZEN)in feeds in China between 2021 and 2024.A total of 23,003 feed samples,including 17,489 feedstuff samples and 5,514 complete feed samples,were collected from different provinces of China for mycotoxin analysis.Results The analyzed mycotoxins displayed considerably high contamination in the feed samples,with the individual contamination of AFB_(1),DON,and ZEN were 20.0%–100%,33.3%–100%,and 85.0%–100%,respectively.The average concentrations of AFB_(1),DON,and ZEN were 1.2–728.7μg/kg,106–8,634.8μg/kg,and 18.1–3,341.6μg/kg,respectively.Notably,the rates over China’s safety standards for AFB_(1),DON,and ZEN in raw ingredients were 9.7%,2.7%,and 15.7%,respectively.Meanwhile,3.5%,1.1%,and 8.7%of analyzed complete feeds exceeded China’s safety standards for AFB_(1),DON,and ZEN,respectively.Moreover,the co-contamination rates of AFB_(1),DON,and ZEN in more than 70%of raw ingredients and 87.5%of complete feed products were 60.0%–100%and 61.5%–100%,respectively.Conclusion This study reveals that the feeds in China have commonly been contaminated with AFB_(1),DON,and ZEN alone and their combination during the past four years.These findings highlight the significance of monitoring mycotoxin contaminant levels in domestic animal feed and the importance of carrying out feed administration and remediation strategies for mycotoxin control.
基金supported by the Chinese Natural Science Foundation Projects 32072775,32272915 and 32472949the National Key Research and Development Programs of China(2023YFD1301003 and 2023YFD1301005)the Fundamental Research Funds for the Central Universities(2662023DKPY002)。
文摘Background AFB_(1)-8,9-exo-epoxide(AFBO)is the highly toxic product of Aflatoxin B_(1)(AFB_(1)).Glutathione S-transferases(GSTs)play pivotal roles in detoxifying AFB_(1) by catalyzing the conjugation of AFBO with glutathione(GSH).Although there are over 20 GST isozymes that have been identified in chicken,GST isozymes involved in the detoxification process of AFB_(1) have not been identified yet.The objective of this study was to determine which GST isozymes played key role in detoxification of AFB_(1).Results A total of 17 pcDNA3.1(+)-GST isozyme plasmids were constructed and the GST isozyme genes were overexpressed by 80–2,500,000 folds in the chicken Leghorn male hepatoma(LMH)cells.Compared to the AFB_(1) treatment,overexpression of GSTA2X,GSTA3,GSTT1L,GSTZ1-1,and GSTZ1-2 increased the cell viability by 6.5%–17.0%in LMH cells.Moreover,overexpression of five GST isozymes reduced the release of lactate dehydrogenase and reactive oxygen species by 8.8%–64.4%,and 57.2%–77.6%,respectively,as well as enhanced the production AFBO-GSH by 15.8%–19.6%,thus mitigating DNA damage induced by AFB_(1).After comprehensive evaluation of various indicators,GSTA2X displayed the best detoxification effects against AFB_(1).GSTA2X was expressed in Pichia pastoris X-33 and its enzymatic properties for catalyzing the conjugation of AFBO with GSH showed that the optimum temperature and pH were 20–25℃ and 7.6–8.6 as well as the enzymatic kinetic parameter V_(max) was 0.23 nmol/min/mg and the Michaelis constant was 86.05μmol/L with the AFB_(1) as substrate.Conclusions In conclusion,GSTA2X,GSTA3,GSTT1L,GSTZ1-1,and GSTZ1-2 played key roles in AFB_(1) detoxification,which will provide new remediation strategies to prevent aflatoxicosis in chickens.
基金funded by the Characteristic Innovation Project of the Guangdong Provincial Department of Education(2024 KTSCX198)Guangdong Basic and Applied Basic Research Foundation(2024 A1515012201)Guangdong Feed Industry Technology System(2024 CXTD14).
文摘Background Aflatoxin B_(1)(AFB_(1))risks animal and human health,and the liver is considered the most crucial detoxification organ.Phlorotannin(PT)is a polyhydroxy phenol that has a wide range of biological activities,including antioxidation and hepatoprotection,which can promote the ability of liver detoxification.This study aimed to elucidate the protective effect of PT on AFB_(1)-induced liver damage in broilers.Results In vivo experiment showed that the PT reduced AFB_(1) content and AFB_(1)-exo-8,9-epoxide DNA(AFBODNA)concentration in serum and liver(P<0.05),improved the histomorphology of liver and hepatic mitochondria,and activated nuclear factor erythroid 2-related factor 2(Nrf2)-related antioxidant and detoxification pathway by upregulating the activities of antioxidant enzymes(catalase[CAT],glutathione S-transferase[GST])and total antioxidant capacity(T-AOC)level(P<0.05),and inhibited the mRNA expression of CYP1A1(cytochrome P450 family 1 subfamily A member 1)and phase Ⅱ detoxification enzyme related genes(GPX1,GSTT1,and NQO1)of broilers exposed to AFB_(1)(P<0.05).Meanwhile,PT upregulated the Nrf1 pathway-related mitochondrial biosynthetic genes(Nrf1,mitochondrial transcription factor A[TFAM],mitofusin 1[MFN1])in broilers fed AFB_(1) contaminated diet(P<0.05).In vitro verification study suggested that the use of Nrf2/Nrf1 inhibitors suppressed the ameliorative role of PT on AFB_(1)-induced liver injury of broilers,which was manifested in the mRNA expression of Nrf2,NQO1,GSTT3,Nrf1,TFAM,and other genes decreasing(P<0.05),and down-regulation of the protein expression of Nrf2,total and nucleus p-Nrf2,and total and nucleus p-Nrf1(P<0.05).Conclusion The PT ameliorates oxidative stress and hepatotoxicity by activating the Nrf2-mediated phase Ⅱ detoxification enzymes pathway and maintains mitochondrial homeostasis by activating the Nrf1 signaling pathway in broilers exposed to AFB_(1).
基金supported by the Scientific and Technological Project of Henan Province(232102321117)National Natural Science Foundation of China(82202198)+2 种基金the National Engineering Research Center of Wheat and Corn Further Processing of Henan University of Technology(NL2022010)Project of Basic Research Fund of Henan Provincial Institute of Medical and Pharmacological Sciences(2023BP0106)the Innovative Funds Plan of Henan University of Technology(2020ZKCJ23)。
文摘Aflatoxin B1(AFB1)is a carcinogenic toxin naturally produced in most food crops that severely threaten human health,and effective methods are urgent to improve the detection accuracy.Herein an indirect competitive immunosorbent approach was elaborately developed based on high-affinity immunoglobulin G(IgG)coupled CuO-anchored Fe_(3)O_(4)nanozymes for precise and ultrasensitive detection of AFB_(1)in food crops including peanut,corn and wheat.The high-affinity nanozymes were fabricated by the assembly of inner core Fe_(3)O_(4)nanoparticles and mesoporous silica capping layer,Cu O further situated within large aperture of the coating layer via in-situ growth,and then conjugated with ligand rabbit anti-mouse Ig G,which can specifically bind with AFB_(1).The results showed the hybrid high-affinity nanozymes displayed enhanced peroxidasemimic activities and catalytic performances,achieving a linear range of 0.06-61.93(lg(ng/mL))and a detection limit of 0.0037 ng/mL,30 times better than that of the conventional enzyme-linked immunosorbent assay.The constructed nanozymes were successfully applied to the detection of AFB_(1)in food products with an average spiked recovery of 96.53%and relative standard deviations less than 2.8%.Therefore,the accurate hybrid nanozymes may serve for AFB_(1)detection in various foods in future.
基金supported by grant from the Science and Technology Program of Zhejiang Province 2022C04034(to Jinzhi Zhang,Junli Zhu and Haifeng Wang)the Key Research and Development Program of China 2022YFD1300602(to Haifeng Wang)。
文摘Background Aflatoxins have been reported as a significant pollutant in feed,capable of causing harm to the liver,gastrointestinal tract and kidneys of piglets.However,research on the interactions among aflatoxin B1(AFB1),bile acid(BA)metabolism and gut microbiota is limited.Methods In this study,piglets were treated with AFB1 and antibiotics(ABX)to evaluate the interaction between AFB1 and gut microbiota.Subsequently,the roles of the farnesoid X receptor(FXR)and sterol 12α-hydroxylase(CYP8B1)in AFB1 absorption were studied by using FXR agonists obeticholic acid(OCA)and Cyp8b1-knockout(KO)mice,respectively.Result AFB1 inhibited bile salt hydrolase(BSH)activity in ileal microbiota,downregulated ileal FXR expression,and upregulated CYP8B1 expression in liver,increasing the proportion of 12α-OH BAs and potentially enhancing AFB1 absorption.ABX treatment reduced AFB1 absorption and liver damage,and unexpectedly increased BSH activity,counteracting the AFB1-induced downregulation of FXR and upregulation of CYP8B1.OCA reactivated ileal FXR,reduced AFB1 absorption,and alleviated liver damage.Furthermore,Cyp8b1-KO mice showed increased resistance to AFB1-induced liver damage by lowering AFB1 absorption.Conclusions These results underscore the significance of gut microbiota and BAs in AFB1 absorption,suggesting new strategies to mitigate health risks from AFB1 in piglets.
基金Supported by the Chongqing Health Commission and Science and Technology Bureau,No.2023MSXM060.
文摘Hepatocellular carcinoma(HCC)is a major global health challenge,particularly in regions with high aflatoxin B1(AFB1)exposure.This editorial explores the mechanistic interplay between AFB1 and tissue inhibitor of metalloproteinase-3(TIMP-3)in AFB1-related HCC.TIMP-3,frequently downregulated in HCC due to promoter methylation,is linked to increased tumor aggressiveness and poor prognosis.We propose that AFB1 induces epigenetic silencing of TIMP-3,potentially via DNA adducts and oxidative stress,exacerbating AFB1-related HCC progression.This AFB1-TIMP-3 axis highlights TIMP-3’s potential as a prognostic biomarker and therapeutic target.Future research should focus on elucidating these molecular pathways and integrating TIMP-3 into clinical practice for early detection and targeted therapies in AFB1-prevalent regions.
基金financially supported by China Postdoctoral Science Foundation(No.2022T150708)National Key Research and Development Program of China(No.2023YFF1104600)National Natural Science Foundation of China(Nos.32072305,32102089)。
文摘Aflatoxins B1(AFB1)contamination in agro-food holds great threaten to human and animal health.Conventional test strips for rapid AFB1 visualized monitoring remains challenged by improvement of sensitivity and matrix interference resistance.In this case,we developed a portable electrochemiluminescence(ECL)imaging test strip with dual-signal outputs for AFB1 quantification in corn samples.RuPEI@SiO_(2)@Au nanospheres were synthesized for bonding with anti-AFB1 antibody and then colorimetrical signal-reported on test line through the capillary flow at strips.Meanwhile,ECL imaging signal of the constructed carbon-ink-based working electrode on polyvinyl chloride substrate of strips was exported under an applied potential of 1.25 V.The whole ECL test strips not only endowed convenient colorimetric responses but guaranteed quick-witted ECL image distinguishment even at extremely low AFB1 content.The detection limit of this ECL imaging-integrated mode was 10-fold lower than that of only colorimetric mode.Furthermore,satisfactory selectivity,reliability and practicability of the as-proposed ECL test strips were demonstrated.This work offered a promising platform for on-site,accurate and sensitive detection of pollutants in foods.
基金supported by National Key Research and Development Program of China (2021YFF0601902)Shanxi Scholarship Council of China (2021-068)+1 种基金Opening Project of Key Laboratory of Detection for Mycotoxins, Ministry of Agriculture and Rural Affairs China (SWDSJC2021001)Shanxi Agricultural University High-Level Talent Project (2021XG013)。
文摘Given severe health-hazardous effects of aflatoxin B1(AFB1) widely occurring in cereal grains and animal feeds,it is highly urgent to develop analytical methods for its rapid screening.In this work,we proposed a simple and high-throughput method for the determination of AFB1 in millet and buckwheat samples using high performance thin layer chromatography(HPTLC) linked to fluorescence densitometry.The first step was to optimize the solid-liquid extraction for the crude clean-up of the samples.The QuEChERS(Quick,Easy,Cheap,Effective,Robust and Safe) extraction strategy was used and different solvent systems for their extraction efficiency of AFB1 from the samples were evaluated.Then,trichloromethane:ethyl acetate(7:3,V/V) was used as the mobile phase to realize the separation of the targeted compound from background noises on silica gel plates.Quantification was readily performed with densitometry in fluorescence mode.In order to fix the optimal excitation wavelength,spectra scanning ranging 250-400 nm was carried out,revealing that 364 nm light gave the highest signal.With the optimized optical system,high sensitivity to AFB1 was achieved,with a limit of detection(LOD) at 3 μg/kg.Apart from that,good linearity(0.999) was obtained within the range of 1-80 ng/band of AFB 1.To assess the analysis accuracy,2 levels of AFB 1(50 and100 μg/kg) were spiked into real grain samples.The obtained results showed that the recovery rates were within the range of 81.6%-114.0%.The proof-of-concept results of this work evidenced that HPTLC is a promising analytical tool for the screening of mycotoxin in difficult samples.
