Two cotton research institute(CRI)near-isogenic lines,CRI-12 glanded and CRI-12 glandless,were used to pinpoint potential genes and metabolic pathways linked to gossypol biosynthesis through transcriptome sequencing.W...Two cotton research institute(CRI)near-isogenic lines,CRI-12 glanded and CRI-12 glandless,were used to pinpoint potential genes and metabolic pathways linked to gossypol biosynthesis through transcriptome sequencing.We discovered more than 235 million clean reads and 1,184 differentially expressed genes(DEGs).Consecutively,we conducted a weighted gene co-expression network analysis and found a strong correlation between white and yellow modules containing GhTPS(GH_D09G0090)and GhCYP(GH_D05G2016)hub genes with the gossypol content.Importance of the GhTPS and GhCYP genes was demonstrated using RT-qPCR,virusinduced gene silencing(VIGS),and target metabolite analysis.Silencing these genes resulted in fewer glands on both leaves and stems two weeks after the infection compared to the wild type.In addition,152 metabolites were identified through targeted metabolite profiling.Differential metabolite screening revealed 12 and 18 significantly different metabolites in TRV:GhTPS and TRV:GhCYP plants vs.the control group,respectively,showing a reduction in the accumulation of metabolites compared to the control.Content of hemigossypol,the final product of gossypol biosynthesis,was also reduced,as revealed by target metabolite analysis,suggesting the role of these genes in the gossypol biosynthetic pathway.Furthermore,a highly significant difference in gossypol content between the glanded and glandless lines was recorded.Findings of this study reveal a strong link between the gossypol content and GhTPS and GhCYP hub genes,suggesting their role in the gossypol biosynthetic pathway to reduce the accumulation of hemigossypol,which may offer new comprehension into the regulatory checkpoints of the gossypol biosynthesis pathway in cotton.展开更多
Background Intestinal barrier is a dynamic interface between the body and the ingested food components, however, dietary components or xenobiotics could compromise intestinal integrity, causing health risks to the hos...Background Intestinal barrier is a dynamic interface between the body and the ingested food components, however, dietary components or xenobiotics could compromise intestinal integrity, causing health risks to the host. Gossypol, a toxic component in cottonseed meal(CSM), caused intestinal injury in fish or other monogastric animals. It has been demonstrated that probiotics administration benefits the intestinal barrier integrity, but the efficacy of probiotics in maintaining intestinal health when the host is exposed to gossypol remains unclear. Here, a strain(YC) affiliated to Pediococcus pentosaceus was isolated from the gut of Nile tilapia(Oreochromis niloticus) and its potential to repair gossypol-induced intestinal damage was evaluated.Results A total of 270 Nile tilapia(2.20 ± 0.02 g) were allotted in 3 groups with 3 tanks each and fed with 3 diets including CON(control diet), GOS(control diet containing 300 mg/kg gossypol) and GP(control diet containing 300 mg/kg gossypol and 10^(8) colony-forming unit(CFU)/g P. pentosaceus YC), respectively. After 10 weeks, addition of P. pentosaceus YC restored growth retardation and intestinal injury induced by gossypol in Nile tilapia. Transcriptome analysis and si RNA interference experiments demonstrated that NOD-like receptors(NLR) family caspase recruitment domain(CARD) domain containing 3(Nlrc3) inhibition might promote intestinal stem cell(ISC) proliferation, as well as maintaining gut barrier integrity. 16S r RNA sequencing and gas chromatography-mass spectrometry(GC-MS) revealed that addition of P. pentosaceus YC altered the composition of gut microbiota and increased the content of propionate in fish gut. In vitro studies on propionate's function demonstrated that it suppressed nlrc3 expression and promoted wound healing in Caco-2 cell model.Conclusions The present study reveals that P. pentosaceus YC has the capacity to ameliorate intestinal barrier injury by modulating gut microbiota composition and elevating propionate level. This finding offers a promising strategy for the feed industry to incorporate cottonseed meal into fish feed formulations.展开更多
BACKGROUND Leukemia stem cells(LSCs)are found to be one of the main factors contributing to poor therapeutic effects in acute myeloid leukemia(AML),as they are protected by the bone marrow microenvironment(BMM)against...BACKGROUND Leukemia stem cells(LSCs)are found to be one of the main factors contributing to poor therapeutic effects in acute myeloid leukemia(AML),as they are protected by the bone marrow microenvironment(BMM)against conventional therapies.Gossypol acetic acid(GAA),which is extracted from the seeds of cotton plants,exerts anti-tumor roles in several types of cancer and has been reported to induce apoptosis of LSCs by inhibiting Bcl2.AIM To investigate the exact roles of GAA in regulating LSCs under different microenvironments and the exact mechanism.METHODS In this study,LSCs were magnetically sorted from AML cell lines and the CD34+CD38-population was obtained.The expression of leucine-rich pentatricopeptide repeat-containing protein(LRPPRC)and forkhead box M1(FOXM1)was evaluated in LSCs,and the effects of GAA on malignancies and mitochondrial RESULTS LRPPRC was found to be upregulated,and GAA inhibited cell proliferation by degrading LRPPRC.GAA induced LRPPRC degradation and inhibited the activation of interleukin 6(IL-6)/janus kinase(JAK)1/signal transducer and activator of transcription(STAT)3 signaling,enhancing chemosensitivity in LSCs against conventional chemotherapies,including L-Asparaginase,Dexamethasone,and cytarabine.GAA was also found to downregulate FOXM1 indirectly by regulating LRPPRC.Furthermore,GAA induced reactive oxygen species accumulation,disturbed mitochondrial homeostasis,and caused mitochondrial dysfunction.By inhibiting IL-6/JAK1/STAT3 signaling via degrading LRPPRC,GAA resulted in the elimination of LSCs.Meanwhile,GAA induced oxidative stress and subsequent cell damage by causing mitochondrial damage.CONCLUSION Taken together,the results indicate that GAA might overcome the BMM protective effect and be considered as a novel and effective combination therapy for AML.展开更多
Cottonseed meal(CM) was used at up to 36.95% content in the diet(replacing 60% of dietary fish meal protein) without any negative effects on growth performance of pre-adult grass carp(initial body weight,761 g) under ...Cottonseed meal(CM) was used at up to 36.95% content in the diet(replacing 60% of dietary fish meal protein) without any negative effects on growth performance of pre-adult grass carp(initial body weight,761 g) under outdoor conditions.A culture trial was conducted in net cages installed in a large concrete pond.Seven isonitrogenous and isoenergetic diets containing a gradient of CM concentrations(0,12.2%,24.4%,36.6%,48.8%,54.8%,and 61.0%) as replacement for dietary fish meal protein(0,20%,40%,60%,80%,90%,and 100%) were formulated.Dietary non-resistant starch(from maize) was inverse to dietary CM.Growth performance and feed utilization of fish fed the diets containing CM replacing 0-40% fishmeal protein were not affected after the 6-week feeding trial.Accumulation of hepatopancreatic total gossypol in the hepatopancreas was significantly correlated with free gossypol content in the diets(HTG=88.6+1.5×DFG,R^2=0.89,P<0.05).Intestinal a-amylase and γ-glutamyl transpeptidase activities rose along with increasing dietary CM level.The structure of the mid-intestinal tissues and the ultrastructure of the enterocyte microvilli were normal when dietary CM was <36.6%(60% protein replacement).Increasing dietary CM content increased serum alanine aminotransferase levels but decreased serum alkaline phosphatase,cholesterol,high-density lipoprotein cholesterol,low-density lipoprotein cholesterol,triglycerides,and albumin(P<0.05).展开更多
To investigate the structural form of gossypol and gossypolone Schiff＇s bases, seven relevant Schiff＇s bases were synthesized and the electrospray ionization-tandem mass spectrometry （ESI-MS/MS） with low-energy co...To investigate the structural form of gossypol and gossypolone Schiff＇s bases, seven relevant Schiff＇s bases were synthesized and the electrospray ionization-tandem mass spectrometry （ESI-MS/MS） with low-energy collision-induced dissociation was used to analyze their fragmentations. A common fragmentation pathway with the loss of RNH2 from those schiff＇s bases quasi-molecular ions was observed and proposed on the basis of their MS/MS spectra data. This common pathway indicated that those Schiff＇s bases existed mainly as the enamine form not the imine form previously showed in most reports.展开更多
The objective of this work was to investigate the effect of six individual strains of fungi on the reduction of gossypol levels and nutritional value during solid substrate fen＇aentation of cottonseed meal （CSM）. S...The objective of this work was to investigate the effect of six individual strains of fungi on the reduction of gossypol levels and nutritional value during solid substrate fen＇aentation of cottonseed meal （CSM）. Six groups of disinfected CSM substrate were incubated for 48 h after inoculation with either of the fungi C. capsuligena ZD- 1, C. tropicalis ZD-3, S. cerevisae ZD-5, A. terricola ZD-6, A. oryzae ZD-7, or A. niger ZD-8. One not inoculated group （substrate） was used as a control. Levels of initial and final free gossypol （FG）, crude protein （CP）, amino acids （AA） and in vitro digestibility were assayed. The experiment was done in triplicate. The experimental results indicated that microbial fermentation could greatly decrease （P〈0.05） FG levels in CSM. The detoxification efficiency differed between the species of microorganisms applied. From the perspective of reducing CSM potential toxicity, C. tropicalis ZD-3 was most successful followed by S. cerevisae ZD-5 and A. niger ZD-8. They could reduce FG levels of CSM to 29.8, 63.07 and 81.50 mg/kg based on DM （dry matter）, respectively, and their detoxification rates were 94.57%, 88.51% and 85.16%, respectively. If crude protein, amino acids content and their in vitro digestibility were also taken into account, Aniger ZD-8 may be the best choice. The CP content of CSM substrate fermented by C. tropicalis ZD-3 and A. niger ZD-8 were improved by 10.76% and 22.24%; the TAA （total amino acids） contents were increased by 7.06% and 11.46%, and the EAA （essential amino acids） were raised by 7.77% and 12.64%, respectively. Especially, the levels of methionine, lysine and threonine were improved greatly （P〈0.05）. The in vitro CP digestibility of CSM fermented by C. tropicalis ZD-3 and A. niger ZD-8 was improved by 13.42% and 18.22%, the TAA were increased by 17.75% and 22.88%, and the EAA by 16.61% and 21.01%, respectively. In addition, the in vitro digestibility of methionine, lysine and threonine was also improved greatly （P〈0.05）.展开更多
Aim: To ascertain whether the side effects of gossypol, hypokalemia and irreversibility, could be avoided on dose re-duction. Methods: Seventy-seven male volunteers were divided into 3 groups: control (22 cases), 10 m...Aim: To ascertain whether the side effects of gossypol, hypokalemia and irreversibility, could be avoided on dose re-duction. Methods: Seventy-seven male volunteers were divided into 3 groups: control (22 cases), 10 mg gossypol(29 cases) and 12.5 mg (26 cases). Serum levels of testosterone, FSH and LH were measured by RIA and potassiumby flame photometry. Spema counts and motility were examined before and regularly after treatment for the evaluationof contraceptive efficacy. Results: The average sperm density and motility started to decrease significantly by theend of month 2 of medication and gradually reached the infertility levels ( < 4 million /mL) in both treated groups. Af-ter that the 10 mg group was asked to take the same dose every other day for up to a total observation period of 16-18months for the maintenance of infertility. Subjects in the 12.5 mg group did not take gossypol any more so as to ob-serve the length of the loading dose required, but in a few, a maintenance dose of 12.5 mg every other day was insti-tuted for a few more months. In both treated groups, none of the spouses was pregnant during the maintenance dose pe-riod. Serum levels of potassium, FSH, LH and testosterone were not significantly changed and not a single volunteercomplained of myoasthenia. After cessation of drug administratioin, the semen data returned to pretreatment levels.Conclusion: A regimen with 10 or 12.5 mg of gossypol as the daily loading dose and 35 or 43.75 mg as the week-ly maintenance dose could induce infertility in male volunteers without developing hypokalemia or irreversibility.(Asian J Androl 2000 Dec; 2: 283-287)展开更多
A series of novel gossypol derivatives were synthesized and screened for their in vitro anti-HIV- 1I activity. The results showed that replacing the aldehyde groups of gossypol with certain oligopeptides and Dglucosam...A series of novel gossypol derivatives were synthesized and screened for their in vitro anti-HIV- 1I activity. The results showed that replacing the aldehyde groups of gossypol with certain oligopeptides and Dglucosamine not only reduced the cytotoxicity of gossypol derivatives but also enhanced their antiviral activity against HIV-1. Interestingly, D-glucosamine derivative of gossypol that lacked the COONa group also exhibited the same potent anti-HIV-1 activity as oligopeptide derivatives with the COONa group. These compounds blocked the entry of HIV-1ⅢB into target cell. which was similar to T20. Furthermore, the molecular docking analysis rationalized their anti-HIV-1 activity. The results also implied that certain oligopeptides and D-glucosamine were important moities to prepare gossypol derivatives as HIV- 1 entry inhibitors besides certain amino acids.展开更多
Aim： To evaluate the anti-proliferative activity and mitochondrial toxicity of gossypol in endometrioma cells maintained in short-term cultures. Methods： （A） Three endometrioma cell lines from patients were treate...Aim： To evaluate the anti-proliferative activity and mitochondrial toxicity of gossypol in endometrioma cells maintained in short-term cultures. Methods： （A） Three endometrioma cell lines from patients were treated with 25 or 50 nmol/L gossypol for up to 12 days. The effect of gossypol on the cell growth was recorded. （B） A phosphorescence oxygen analyzer was used to determine the effects of gossypol on mitochondrial oxygen consumption of six endometrioma cell lines from patients. （C） Cellular gossypol accumulations in three endometrioma cell lines from patients were measured by high-pressure liquid chromatography. Results： Proliferation of the endometrioma cells was inhibited by 25 and 50 nmol/L gossypol. Respiration of the endometrioma cells was inhibited by 10 μmol/L gossypol. Cellular gossypol was detected in the endometrioma cell lines that were treated for 24 h with l0 and 0.3 μmol/L gossypol. Conclusion： Gossypol invokes a potent toxicity on cultured endometrioma cells.展开更多
This work reports the single-molecule detection of gossypol by flow injection analysis with chemiluminescence method. The method is based on the reaction of luminol with ferricyanid in sodium hydroxide medium sensitiz...This work reports the single-molecule detection of gossypol by flow injection analysis with chemiluminescence method. The method is based on the reaction of luminol with ferricyanid in sodium hydroxide medium sensitized by gossypol. Under the optimum conditions, the CL intensity is proportional to the concentration of gossypol over the range of 1.11×10^-17-2.78×10^-16 mol/L in acid solution and 8.00×10^-11-7.39×10^-8mol/L in neutral solution with correlation coefficients 0.9983 and 0.9905, respectively. The detection limits is 1.60×10^-18 mol/L （S/N=3）. The proposed method has been applied for the determination of the gossypol in cottonseeds and pharmaceutical formulations with satisfactory results.展开更多
We investigated the antiproliferative activity of （-）-gossypol on the human prostate cancer cell line PC3 in vitro and in vivo to elucidate its potential molecular mechanisms. Cell growth and viability were evaluate...We investigated the antiproliferative activity of （-）-gossypol on the human prostate cancer cell line PC3 in vitro and in vivo to elucidate its potential molecular mechanisms. Cell growth and viability were evaluated using the 3-（4,5-dimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide （MTT） assay, and cell apoptosis was detected by flow cytometry, terminal deoxynucleotidyl transferase dUTP nick end labelling （TUNEL） and electron microscopy. Expression of proliferating cell nuclear antigen （PCNA）, Bcl-2, CD31, caspase-3 and caspase-8 in tumour tissue was determined by immunohistochemistry. The drug concentration that yielded 50% cell inhibition （IC50 value） was 4.74 μg mL-1. In the PC-3 tumour xenograft study, （-）-gossypol （〉 5 mg kg-1） given once a day for 7 days significantly inhibited tumour growth in a dose-dependent manner. Immunohistochemical analysis revealed that （-）-gossypol enhanced caspase-3 and caspase-8 expression and decreased the expression of PCNA, Bcl-2 and CD31 in tumour tissues. It suggested that cell apoptosis and inhibition of angiogenesis might contribute to the anticancer action of （-）-gossypol.展开更多
[Objective] This study aims to reduce the free gossypol(FG) and improve utilization rate of cottonseed meal(CSM) by solid state fermentation(SSF). [Method]Bacillus subtilis GJ00141 and Saccharomyces cerevisiae GJ00079...[Objective] This study aims to reduce the free gossypol(FG) and improve utilization rate of cottonseed meal(CSM) by solid state fermentation(SSF). [Method]Bacillus subtilis GJ00141 and Saccharomyces cerevisiae GJ00079 were applied for the SSF and the optimal carbon source, nitrogen source, inorganic salt, moisture content, inoculum level, fermentation temperature, and fermentation time were investigated. The detoxifying effects of different products of GJ00141 were examined with gossypol as substrate. A total of 90 one-day-old broilers were randomized into group A [control, basal diet with 36% soybean meal(SM)], group B(basal diet with 18% SM and 18% CSM), and group C [basal diet with 18% SM and 18% fermented CSM(FCSM)] and thereby the influence of FCSM on the growth of broilers was explored. [Results] The maximum reduction rate(59%) of FG was achieved under the following fermentation conditions: solid medium composed of 96% CSM, 1%glucose, 1% ammonium sulfate, and 2% corn grits, 45% moisture content, 20%inoculum, fermentation at 30 °C for 60 h. Both the viable and inactivated cells of GJ00141 can reduce the content of gossypol, but the reduction rates were only about 20% after 72 h of incubation. Cellular contents and supernatant demonstrated strong detoxifying activity, which achieved the reduction rates of about 95% after 48 h, and the removal was free from the influence of proteinase K, heat, or EDTA. In the 42 d feeding experiment on broilers, the ratios of feed to gain were insignificantly different between the group C and group A. [Conclusion] This method achieved high rate of removing FG in CSM. The reason was the likelihood that the stable compounds in the cellular contents and supernatant of GJ00141 adsorb or bind to FG. Broilers grew well with the FCSM. Thus, it was an efficient detoxifying method for CSM.展开更多
A pharmacokinetic comparison of gossypol isomers in cattle was made by investigating their transfer from ingested food to plasma and their degradation by rumen microbes. The gossypol isomers in whole cottonseed, plasm...A pharmacokinetic comparison of gossypol isomers in cattle was made by investigating their transfer from ingested food to plasma and their degradation by rumen microbes. The gossypol isomers in whole cottonseed, plasma, and rumen fluid were determined by high-performance liquid chromatography(HPLC). The intakes of(+)-and(-)-gossypol by cows on three farms were about 5.6–8.5 and 3.8–5.9 g/(d·herd), respectively. The plasma gossypol concentrations increased as ingestion increased and ranged from 0.31 to 0.48 μg/ml for the(+) form and from 0.39 to 0.59 μg/ml for the(-) form. The(+) form was slightly predominant(58.8%–59.8%) in the gossypol ingested by the cows, whereas the(-) form predominated(54.6%–55.9%) in the plasma. An in vitro study showed that at 6 h, 67.4% and 85.7% of the(-)-gossypol were degraded in 500 and 1000 μg/g treatment groups, respectively, and these increased to 83.6% and 92.5%, respectively, at 12 h. The regularity of the degradation of(+)-gossypol was similar to that of(-)-gossypol. These results showed that(-)-gossypol may be more persistent than(+)-gossypol in plasma, and that the degradation of the gossypol isomers in the rumen is rapid and not enantioselective in cattle.展开更多
Background:Gossypol found in cottonseeds is toxic to human beings and monogastric animals and is a primary parameter for the integrated utilization of cottonseed products.It is usually determined by the techniques rel...Background:Gossypol found in cottonseeds is toxic to human beings and monogastric animals and is a primary parameter for the integrated utilization of cottonseed products.It is usually determined by the techniques relied on complex pretreatment procedures and the samples after determination cannot be used in the breeding program,so it is of great importance to predict the gossypol content in cottonseeds rapidly and nondestructively to substitute the traditional analytical method.Results:Gossypol content in cottonseeds was investigated by near-infrared spectroscopy(NIRS)and high-performance liquid chromatography(HPLC).Partial least squares regression,combined with spectral pretreatment methods including Savitzky-Golay smoothing,standard normal variate,multiplicative scatter correction,and first derivate were tested for optimizing the calibration models.NIRS technique was efficient in predicting gossypol content in intact cottonseeds,as revealed by the root-mean-square error of cross-validation(RMSECV),root-mean-square error of prediction(RMSEP),coefficient for determination of prediction(R_(p)^(2)),and residual predictive deviation(RPD)values for all models,being 0.05∼0.07,0.04∼0.06,0.82∼0.92,and 2.3∼3.4,respectively.The optimized model pretreated by Savitzky-Golay smoothing+standard normal variate+first derivate resulted in a good determination of gossypol content in intact cottonseeds.Conclusions:Near-infrared spectroscopy coupled with different spectral pretreatments and partial least squares(PLS)regression has exhibited the feasibility in predicting gossypol content in intact cottonseeds,rapidly and non destructively.It could be used as an alternative method to substitute for traditional one to determi ne the gossypol content in intact cottonseeds.展开更多
The objective was to evaluate the toxicity effect of gossypol on ultrastructure of mouse testis and the expression of Bax mRNA and Bcl-2 mRNA of sperm cells in mice. Forty-eight male mice were randomly divided into fo...The objective was to evaluate the toxicity effect of gossypol on ultrastructure of mouse testis and the expression of Bax mRNA and Bcl-2 mRNA of sperm cells in mice. Forty-eight male mice were randomly divided into four groups： control group, L-group （30 mg-kgt. d）, M-group （60 mg·kg-1 ·d） and H-group （120 mg·kg-1· d） and were orally administrated with gossypol diluted by sodium carboxymethyl cellulose （SCC） or SCC （control group） for 20 days. On the 21st day, all the mice were killed and ultrastructure changes of testis were observed by TEM. mRNA expression of Bax and Bcl-2 in testis was measured by semiquantitative RT-PCR. The results showed that the testicular ultrastructure in three treated groups was gradually damaged, according to the dosage of gossypol and cellular structure disordered and organdie degenerated, manifesting vacuolation of mitochondria, expansion of endoplasmie reticulum, mRNA expression of Bcl-2 in testis significantly increased （p〈0.05） in L-group and then significantly decreased （p〈0.05, p〈0.01） in M-group and H-group compared with that in the control group; mRNA expression of Bcl-2 in M-group and H-group significantly decreased （p〈0.05, p〈0.01） than that in L-group and Bcl-2 mRNA expression in H-group showed a significant decrease （p〈0.05） compared with that in M-group. On the other hand, mRNA expression of Bax significant increased （p〈0.05,p〈0.01） in M-group and H-group than that in the control group. The ratio of Bcl-2/Bax significantly reduced 07〈0.05, p〈0.01） in the treated group than that in the control group and was found to be an obvious dose-dependent. It demonstrated that the gnssypol could induce the changes on ultrastructure of mice testis, down-regulate mRNA expression of Bcl-2 and up-regnlate mRNA expression of Bax, which indicated that sperm ceils were induced apoptosis.展开更多
基金financial help from the National Key R&D Program of China(2021YFE0101200)the Key Research and Development Project of Jiangsu Province,China(Modern Agriculture,BE2022364)+1 种基金the State Key Laboratory of Cotton Bio-breeding and Integrated Utilization Open Fund,China(CB2024A06)support of the Ministry of Science,Technological Development and Innovation of the Republic of Serbia(451-03-66/2024-03/200007)。
文摘Two cotton research institute(CRI)near-isogenic lines,CRI-12 glanded and CRI-12 glandless,were used to pinpoint potential genes and metabolic pathways linked to gossypol biosynthesis through transcriptome sequencing.We discovered more than 235 million clean reads and 1,184 differentially expressed genes(DEGs).Consecutively,we conducted a weighted gene co-expression network analysis and found a strong correlation between white and yellow modules containing GhTPS(GH_D09G0090)and GhCYP(GH_D05G2016)hub genes with the gossypol content.Importance of the GhTPS and GhCYP genes was demonstrated using RT-qPCR,virusinduced gene silencing(VIGS),and target metabolite analysis.Silencing these genes resulted in fewer glands on both leaves and stems two weeks after the infection compared to the wild type.In addition,152 metabolites were identified through targeted metabolite profiling.Differential metabolite screening revealed 12 and 18 significantly different metabolites in TRV:GhTPS and TRV:GhCYP plants vs.the control group,respectively,showing a reduction in the accumulation of metabolites compared to the control.Content of hemigossypol,the final product of gossypol biosynthesis,was also reduced,as revealed by target metabolite analysis,suggesting the role of these genes in the gossypol biosynthetic pathway.Furthermore,a highly significant difference in gossypol content between the glanded and glandless lines was recorded.Findings of this study reveal a strong link between the gossypol content and GhTPS and GhCYP hub genes,suggesting their role in the gossypol biosynthetic pathway to reduce the accumulation of hemigossypol,which may offer new comprehension into the regulatory checkpoints of the gossypol biosynthesis pathway in cotton.
基金supported by the Provincial Science and Technology Innovative Program for Carbon Peak and Carbon neutrality of Jiangsu of China (BE2022422)National Natural Science Foundation of China (32373145)。
文摘Background Intestinal barrier is a dynamic interface between the body and the ingested food components, however, dietary components or xenobiotics could compromise intestinal integrity, causing health risks to the host. Gossypol, a toxic component in cottonseed meal(CSM), caused intestinal injury in fish or other monogastric animals. It has been demonstrated that probiotics administration benefits the intestinal barrier integrity, but the efficacy of probiotics in maintaining intestinal health when the host is exposed to gossypol remains unclear. Here, a strain(YC) affiliated to Pediococcus pentosaceus was isolated from the gut of Nile tilapia(Oreochromis niloticus) and its potential to repair gossypol-induced intestinal damage was evaluated.Results A total of 270 Nile tilapia(2.20 ± 0.02 g) were allotted in 3 groups with 3 tanks each and fed with 3 diets including CON(control diet), GOS(control diet containing 300 mg/kg gossypol) and GP(control diet containing 300 mg/kg gossypol and 10^(8) colony-forming unit(CFU)/g P. pentosaceus YC), respectively. After 10 weeks, addition of P. pentosaceus YC restored growth retardation and intestinal injury induced by gossypol in Nile tilapia. Transcriptome analysis and si RNA interference experiments demonstrated that NOD-like receptors(NLR) family caspase recruitment domain(CARD) domain containing 3(Nlrc3) inhibition might promote intestinal stem cell(ISC) proliferation, as well as maintaining gut barrier integrity. 16S r RNA sequencing and gas chromatography-mass spectrometry(GC-MS) revealed that addition of P. pentosaceus YC altered the composition of gut microbiota and increased the content of propionate in fish gut. In vitro studies on propionate's function demonstrated that it suppressed nlrc3 expression and promoted wound healing in Caco-2 cell model.Conclusions The present study reveals that P. pentosaceus YC has the capacity to ameliorate intestinal barrier injury by modulating gut microbiota composition and elevating propionate level. This finding offers a promising strategy for the feed industry to incorporate cottonseed meal into fish feed formulations.
文摘BACKGROUND Leukemia stem cells(LSCs)are found to be one of the main factors contributing to poor therapeutic effects in acute myeloid leukemia(AML),as they are protected by the bone marrow microenvironment(BMM)against conventional therapies.Gossypol acetic acid(GAA),which is extracted from the seeds of cotton plants,exerts anti-tumor roles in several types of cancer and has been reported to induce apoptosis of LSCs by inhibiting Bcl2.AIM To investigate the exact roles of GAA in regulating LSCs under different microenvironments and the exact mechanism.METHODS In this study,LSCs were magnetically sorted from AML cell lines and the CD34+CD38-population was obtained.The expression of leucine-rich pentatricopeptide repeat-containing protein(LRPPRC)and forkhead box M1(FOXM1)was evaluated in LSCs,and the effects of GAA on malignancies and mitochondrial RESULTS LRPPRC was found to be upregulated,and GAA inhibited cell proliferation by degrading LRPPRC.GAA induced LRPPRC degradation and inhibited the activation of interleukin 6(IL-6)/janus kinase(JAK)1/signal transducer and activator of transcription(STAT)3 signaling,enhancing chemosensitivity in LSCs against conventional chemotherapies,including L-Asparaginase,Dexamethasone,and cytarabine.GAA was also found to downregulate FOXM1 indirectly by regulating LRPPRC.Furthermore,GAA induced reactive oxygen species accumulation,disturbed mitochondrial homeostasis,and caused mitochondrial dysfunction.By inhibiting IL-6/JAK1/STAT3 signaling via degrading LRPPRC,GAA resulted in the elimination of LSCs.Meanwhile,GAA induced oxidative stress and subsequent cell damage by causing mitochondrial damage.CONCLUSION Taken together,the results indicate that GAA might overcome the BMM protective effect and be considered as a novel and effective combination therapy for AML.
基金Supported by the Modern Agro-Industry Technology Research System(No.CARS-46-19)the Special Fund for Agro-Scientific Research in the Public Interest(No.201303053)+2 种基金the National Basic Research Program of China(973 Program)(No.2014CB138602)the National Natural Science Foundation of China(No.31101908)the Fund Project in the State Key Laboratory of Freshwater Ecology and Biotechnology(No.2014FBZ04)
文摘Cottonseed meal(CM) was used at up to 36.95% content in the diet(replacing 60% of dietary fish meal protein) without any negative effects on growth performance of pre-adult grass carp(initial body weight,761 g) under outdoor conditions.A culture trial was conducted in net cages installed in a large concrete pond.Seven isonitrogenous and isoenergetic diets containing a gradient of CM concentrations(0,12.2%,24.4%,36.6%,48.8%,54.8%,and 61.0%) as replacement for dietary fish meal protein(0,20%,40%,60%,80%,90%,and 100%) were formulated.Dietary non-resistant starch(from maize) was inverse to dietary CM.Growth performance and feed utilization of fish fed the diets containing CM replacing 0-40% fishmeal protein were not affected after the 6-week feeding trial.Accumulation of hepatopancreatic total gossypol in the hepatopancreas was significantly correlated with free gossypol content in the diets(HTG=88.6+1.5×DFG,R^2=0.89,P<0.05).Intestinal a-amylase and γ-glutamyl transpeptidase activities rose along with increasing dietary CM level.The structure of the mid-intestinal tissues and the ultrastructure of the enterocyte microvilli were normal when dietary CM was <36.6%(60% protein replacement).Increasing dietary CM content increased serum alanine aminotransferase levels but decreased serum alkaline phosphatase,cholesterol,high-density lipoprotein cholesterol,low-density lipoprotein cholesterol,triglycerides,and albumin(P<0.05).
文摘To investigate the structural form of gossypol and gossypolone Schiff＇s bases, seven relevant Schiff＇s bases were synthesized and the electrospray ionization-tandem mass spectrometry （ESI-MS/MS） with low-energy collision-induced dissociation was used to analyze their fragmentations. A common fragmentation pathway with the loss of RNH2 from those schiff＇s bases quasi-molecular ions was observed and proposed on the basis of their MS/MS spectra data. This common pathway indicated that those Schiff＇s bases existed mainly as the enamine form not the imine form previously showed in most reports.
基金Project (No. 30471255) supported by the National Natural ScienceFoundation of China
文摘The objective of this work was to investigate the effect of six individual strains of fungi on the reduction of gossypol levels and nutritional value during solid substrate fen＇aentation of cottonseed meal （CSM）. Six groups of disinfected CSM substrate were incubated for 48 h after inoculation with either of the fungi C. capsuligena ZD- 1, C. tropicalis ZD-3, S. cerevisae ZD-5, A. terricola ZD-6, A. oryzae ZD-7, or A. niger ZD-8. One not inoculated group （substrate） was used as a control. Levels of initial and final free gossypol （FG）, crude protein （CP）, amino acids （AA） and in vitro digestibility were assayed. The experiment was done in triplicate. The experimental results indicated that microbial fermentation could greatly decrease （P〈0.05） FG levels in CSM. The detoxification efficiency differed between the species of microorganisms applied. From the perspective of reducing CSM potential toxicity, C. tropicalis ZD-3 was most successful followed by S. cerevisae ZD-5 and A. niger ZD-8. They could reduce FG levels of CSM to 29.8, 63.07 and 81.50 mg/kg based on DM （dry matter）, respectively, and their detoxification rates were 94.57%, 88.51% and 85.16%, respectively. If crude protein, amino acids content and their in vitro digestibility were also taken into account, Aniger ZD-8 may be the best choice. The CP content of CSM substrate fermented by C. tropicalis ZD-3 and A. niger ZD-8 were improved by 10.76% and 22.24%; the TAA （total amino acids） contents were increased by 7.06% and 11.46%, and the EAA （essential amino acids） were raised by 7.77% and 12.64%, respectively. Especially, the levels of methionine, lysine and threonine were improved greatly （P〈0.05）. The in vitro CP digestibility of CSM fermented by C. tropicalis ZD-3 and A. niger ZD-8 was improved by 13.42% and 18.22%, the TAA were increased by 17.75% and 22.88%, and the EAA by 16.61% and 21.01%, respectively. In addition, the in vitro digestibility of methionine, lysine and threonine was also improved greatly （P〈0.05）.
文摘Aim: To ascertain whether the side effects of gossypol, hypokalemia and irreversibility, could be avoided on dose re-duction. Methods: Seventy-seven male volunteers were divided into 3 groups: control (22 cases), 10 mg gossypol(29 cases) and 12.5 mg (26 cases). Serum levels of testosterone, FSH and LH were measured by RIA and potassiumby flame photometry. Spema counts and motility were examined before and regularly after treatment for the evaluationof contraceptive efficacy. Results: The average sperm density and motility started to decrease significantly by theend of month 2 of medication and gradually reached the infertility levels ( < 4 million /mL) in both treated groups. Af-ter that the 10 mg group was asked to take the same dose every other day for up to a total observation period of 16-18months for the maintenance of infertility. Subjects in the 12.5 mg group did not take gossypol any more so as to ob-serve the length of the loading dose required, but in a few, a maintenance dose of 12.5 mg every other day was insti-tuted for a few more months. In both treated groups, none of the spouses was pregnant during the maintenance dose pe-riod. Serum levels of potassium, FSH, LH and testosterone were not significantly changed and not a single volunteercomplained of myoasthenia. After cessation of drug administratioin, the semen data returned to pretreatment levels.Conclusion: A regimen with 10 or 12.5 mg of gossypol as the daily loading dose and 35 or 43.75 mg as the week-ly maintenance dose could induce infertility in male volunteers without developing hypokalemia or irreversibility.(Asian J Androl 2000 Dec; 2: 283-287)
基金the National Natural Science Foundation of China(No.30770228)for financial support
文摘A series of novel gossypol derivatives were synthesized and screened for their in vitro anti-HIV- 1I activity. The results showed that replacing the aldehyde groups of gossypol with certain oligopeptides and Dglucosamine not only reduced the cytotoxicity of gossypol derivatives but also enhanced their antiviral activity against HIV-1. Interestingly, D-glucosamine derivative of gossypol that lacked the COONa group also exhibited the same potent anti-HIV-1 activity as oligopeptide derivatives with the COONa group. These compounds blocked the entry of HIV-1ⅢB into target cell. which was similar to T20. Furthermore, the molecular docking analysis rationalized their anti-HIV-1 activity. The results also implied that certain oligopeptides and D-glucosamine were important moities to prepare gossypol derivatives as HIV- 1 entry inhibitors besides certain amino acids.
文摘Aim： To evaluate the anti-proliferative activity and mitochondrial toxicity of gossypol in endometrioma cells maintained in short-term cultures. Methods： （A） Three endometrioma cell lines from patients were treated with 25 or 50 nmol/L gossypol for up to 12 days. The effect of gossypol on the cell growth was recorded. （B） A phosphorescence oxygen analyzer was used to determine the effects of gossypol on mitochondrial oxygen consumption of six endometrioma cell lines from patients. （C） Cellular gossypol accumulations in three endometrioma cell lines from patients were measured by high-pressure liquid chromatography. Results： Proliferation of the endometrioma cells was inhibited by 25 and 50 nmol/L gossypol. Respiration of the endometrioma cells was inhibited by 10 μmol/L gossypol. Cellular gossypol was detected in the endometrioma cell lines that were treated for 24 h with l0 and 0.3 μmol/L gossypol. Conclusion： Gossypol invokes a potent toxicity on cultured endometrioma cells.
基金financial support from the National Natural Science Foundation of China(Grant No.20075017)and from Tianjin Normal University.
文摘This work reports the single-molecule detection of gossypol by flow injection analysis with chemiluminescence method. The method is based on the reaction of luminol with ferricyanid in sodium hydroxide medium sensitized by gossypol. Under the optimum conditions, the CL intensity is proportional to the concentration of gossypol over the range of 1.11×10^-17-2.78×10^-16 mol/L in acid solution and 8.00×10^-11-7.39×10^-8mol/L in neutral solution with correlation coefficients 0.9983 and 0.9905, respectively. The detection limits is 1.60×10^-18 mol/L （S/N=3）. The proposed method has been applied for the determination of the gossypol in cottonseeds and pharmaceutical formulations with satisfactory results.
基金This study was supported in part by grants from National Natural Science Foundation of China （No. 30570494 and No. 30772658）. We thank Dr Xing- Bin Hu （The Second Department of Blood Transfusion, Xijing Hospital, Xi＇an, China） for assisting writing this manuscript.
文摘We investigated the antiproliferative activity of （-）-gossypol on the human prostate cancer cell line PC3 in vitro and in vivo to elucidate its potential molecular mechanisms. Cell growth and viability were evaluated using the 3-（4,5-dimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide （MTT） assay, and cell apoptosis was detected by flow cytometry, terminal deoxynucleotidyl transferase dUTP nick end labelling （TUNEL） and electron microscopy. Expression of proliferating cell nuclear antigen （PCNA）, Bcl-2, CD31, caspase-3 and caspase-8 in tumour tissue was determined by immunohistochemistry. The drug concentration that yielded 50% cell inhibition （IC50 value） was 4.74 μg mL-1. In the PC-3 tumour xenograft study, （-）-gossypol （〉 5 mg kg-1） given once a day for 7 days significantly inhibited tumour growth in a dose-dependent manner. Immunohistochemical analysis revealed that （-）-gossypol enhanced caspase-3 and caspase-8 expression and decreased the expression of PCNA, Bcl-2 and CD31 in tumour tissues. It suggested that cell apoptosis and inhibition of angiogenesis might contribute to the anticancer action of （-）-gossypol.
基金Supported by the Academic Research Projects of Beijing Union University (ZK30202104, ZK20202101)。
文摘[Objective] This study aims to reduce the free gossypol(FG) and improve utilization rate of cottonseed meal(CSM) by solid state fermentation(SSF). [Method]Bacillus subtilis GJ00141 and Saccharomyces cerevisiae GJ00079 were applied for the SSF and the optimal carbon source, nitrogen source, inorganic salt, moisture content, inoculum level, fermentation temperature, and fermentation time were investigated. The detoxifying effects of different products of GJ00141 were examined with gossypol as substrate. A total of 90 one-day-old broilers were randomized into group A [control, basal diet with 36% soybean meal(SM)], group B(basal diet with 18% SM and 18% CSM), and group C [basal diet with 18% SM and 18% fermented CSM(FCSM)] and thereby the influence of FCSM on the growth of broilers was explored. [Results] The maximum reduction rate(59%) of FG was achieved under the following fermentation conditions: solid medium composed of 96% CSM, 1%glucose, 1% ammonium sulfate, and 2% corn grits, 45% moisture content, 20%inoculum, fermentation at 30 °C for 60 h. Both the viable and inactivated cells of GJ00141 can reduce the content of gossypol, but the reduction rates were only about 20% after 72 h of incubation. Cellular contents and supernatant demonstrated strong detoxifying activity, which achieved the reduction rates of about 95% after 48 h, and the removal was free from the influence of proteinase K, heat, or EDTA. In the 42 d feeding experiment on broilers, the ratios of feed to gain were insignificantly different between the group C and group A. [Conclusion] This method achieved high rate of removing FG in CSM. The reason was the likelihood that the stable compounds in the cellular contents and supernatant of GJ00141 adsorb or bind to FG. Broilers grew well with the FCSM. Thus, it was an efficient detoxifying method for CSM.
基金Project supported by the National Natural Science Foundation of China(No.31472123)
文摘A pharmacokinetic comparison of gossypol isomers in cattle was made by investigating their transfer from ingested food to plasma and their degradation by rumen microbes. The gossypol isomers in whole cottonseed, plasma, and rumen fluid were determined by high-performance liquid chromatography(HPLC). The intakes of(+)-and(-)-gossypol by cows on three farms were about 5.6–8.5 and 3.8–5.9 g/(d·herd), respectively. The plasma gossypol concentrations increased as ingestion increased and ranged from 0.31 to 0.48 μg/ml for the(+) form and from 0.39 to 0.59 μg/ml for the(-) form. The(+) form was slightly predominant(58.8%–59.8%) in the gossypol ingested by the cows, whereas the(-) form predominated(54.6%–55.9%) in the plasma. An in vitro study showed that at 6 h, 67.4% and 85.7% of the(-)-gossypol were degraded in 500 and 1000 μg/g treatment groups, respectively, and these increased to 83.6% and 92.5%, respectively, at 12 h. The regularity of the degradation of(+)-gossypol was similar to that of(-)-gossypol. These results showed that(-)-gossypol may be more persistent than(+)-gossypol in plasma, and that the degradation of the gossypol isomers in the rumen is rapid and not enantioselective in cattle.
基金The research work was funded by The National Key Technology R&D Program of China(2016YFD0101404)China Agriculture Research System(CARS-18-25)Jiangsu Collaborative Innovation Center for Modern Crop Production.
文摘Background:Gossypol found in cottonseeds is toxic to human beings and monogastric animals and is a primary parameter for the integrated utilization of cottonseed products.It is usually determined by the techniques relied on complex pretreatment procedures and the samples after determination cannot be used in the breeding program,so it is of great importance to predict the gossypol content in cottonseeds rapidly and nondestructively to substitute the traditional analytical method.Results:Gossypol content in cottonseeds was investigated by near-infrared spectroscopy(NIRS)and high-performance liquid chromatography(HPLC).Partial least squares regression,combined with spectral pretreatment methods including Savitzky-Golay smoothing,standard normal variate,multiplicative scatter correction,and first derivate were tested for optimizing the calibration models.NIRS technique was efficient in predicting gossypol content in intact cottonseeds,as revealed by the root-mean-square error of cross-validation(RMSECV),root-mean-square error of prediction(RMSEP),coefficient for determination of prediction(R_(p)^(2)),and residual predictive deviation(RPD)values for all models,being 0.05∼0.07,0.04∼0.06,0.82∼0.92,and 2.3∼3.4,respectively.The optimized model pretreated by Savitzky-Golay smoothing+standard normal variate+first derivate resulted in a good determination of gossypol content in intact cottonseeds.Conclusions:Near-infrared spectroscopy coupled with different spectral pretreatments and partial least squares(PLS)regression has exhibited the feasibility in predicting gossypol content in intact cottonseeds,rapidly and non destructively.It could be used as an alternative method to substitute for traditional one to determi ne the gossypol content in intact cottonseeds.
文摘The objective was to evaluate the toxicity effect of gossypol on ultrastructure of mouse testis and the expression of Bax mRNA and Bcl-2 mRNA of sperm cells in mice. Forty-eight male mice were randomly divided into four groups： control group, L-group （30 mg-kgt. d）, M-group （60 mg·kg-1 ·d） and H-group （120 mg·kg-1· d） and were orally administrated with gossypol diluted by sodium carboxymethyl cellulose （SCC） or SCC （control group） for 20 days. On the 21st day, all the mice were killed and ultrastructure changes of testis were observed by TEM. mRNA expression of Bax and Bcl-2 in testis was measured by semiquantitative RT-PCR. The results showed that the testicular ultrastructure in three treated groups was gradually damaged, according to the dosage of gossypol and cellular structure disordered and organdie degenerated, manifesting vacuolation of mitochondria, expansion of endoplasmie reticulum, mRNA expression of Bcl-2 in testis significantly increased （p〈0.05） in L-group and then significantly decreased （p〈0.05, p〈0.01） in M-group and H-group compared with that in the control group; mRNA expression of Bcl-2 in M-group and H-group significantly decreased （p〈0.05, p〈0.01） than that in L-group and Bcl-2 mRNA expression in H-group showed a significant decrease （p〈0.05） compared with that in M-group. On the other hand, mRNA expression of Bax significant increased （p〈0.05,p〈0.01） in M-group and H-group than that in the control group. The ratio of Bcl-2/Bax significantly reduced 07〈0.05, p〈0.01） in the treated group than that in the control group and was found to be an obvious dose-dependent. It demonstrated that the gnssypol could induce the changes on ultrastructure of mice testis, down-regulate mRNA expression of Bcl-2 and up-regnlate mRNA expression of Bax, which indicated that sperm ceils were induced apoptosis.
