Short-term exposure to gossypol causes reversible reproductive toxicity and nephrotoxicity in mice / 南方医科大学学报

OBJECTIVE@#To study the toxic effects of short-term exposure to gossypol on the testis and kidney in mice and whether these effects are reversible.@*METHODS@#Twenty 7 to 8-week-old male mice were randomized into blank control group, solvent control group, gossypol treatment group and drug withdrawal group. In the former 3 groups, the mice were subjected to daily intragastric administration of 0.3 mL of purified water, 1% sodium carboxymethylcellulose solution, and 30 mg/mL gossypol solution for 14 days, respectively; In the drug withdrawal group, the mice were treated with gossypol solution in the same manner for 14 days followed by treatment with purified water for another 14 days. After the last administration, the mice were euthanized and tissue samples were collected. The testicular tissue was weighed and observed microscopically with HE and PAS staining; the kidney tissue was stained with HE and examined for mitochondrial ATPase activity.@*RESULTS@#Compared with those in the control group, the mice with gossypol exposure showed reduced testicular seminiferous epithelial cells with rounded seminiferous tubules, enlarged space between the seminiferous tubules, interstitium atrophy of the testis, and incomplete differentiation of the spermatogonia. The gossypol-treated mice also presented with complete, non-elongated spermatids, a large number of cells in the state of round spermatids, and negativity for acrosome PAS reaction; diffuse renal mesangial cell hyperplasia, increased mesangial matrix, and adhesion of the mesangium to the wall of the renal capsule were observed, with significantly shrinkage or even absence of the lumens of the renal capsules and reduced kidney mitochondrial ATPase activity. Compared with the gossypol-treated mice, the mice in the drug withdrawal group showed obvious recovery of morphologies of the testis and the kidney, acrosome PAS reaction and mitochondrial ATPase activity.@*CONCLUSIONS@#Shortterm treatment with gossypol can cause reproductive toxicity and nephrotoxicity in mice, but these toxic effects can be reversed after drug withdrawal.

An in vitro and in silico study of anti-dermatophytic activity of gossypol from fruits of Thespesia populnea (L.) Sol. ex Correa

Objective: To isolate, purify, and characterize gossypol from the fruits of Thespesia populnea (L) Sol. ex Correa, test its anti- dermatophytic activity, identify its targets on the dermatophyte, and confirm the binding of gossypol with the fungal target by molecular docking study. Methods: Gossypol from Thespesia populnea was characterized by high performance liquid chromatography, liquid chromatograph- mass spectrometry, Fourier transform infrared spectroscopy, and nuclear magnetic resonance. The anti-dermatophytic activity of gossypol was tested against four different dermatophytes, viz. Trichophyton mentagrophytes, Trichophyton rubrum, Microsporum canis, and Microsporum gypseum. Trichophyton mentagrophytes was selected for further studies. The inhibitory mode of action of gossypol on Trichophyton mentagrophytes was determined by analyzing the modulation of gene expression in various pathways of the dermatophyte. Results: Gossypol inhibited all the dermatophytes. The minimum inhibitory concentrations were 12.5 μg/mL for Trichophyton mentagrophytes and Microsporum canis and 25 μg/mL for Trichophyton rubrum and Microsporum gypseum. The minimum fungicidal concentrations were 50 μg/mL for Trichophyton mentagrophytes, 100 μg/mL for Microsporum canis and Trichophyton rubrum, and 200 μg/mL for Microsporum gypseum. Gossypol inhibited the mRNA expression of metalloprotease (MEP4) and isocitrate lyase (ICL). The binding of gossypol with the enzymes was confirmed by molecular docking studies. The best docking poses were found and the low binding energies were recorded with the two target enzymes. Conclusions: Gossypol is a potential antifungal agent and can be further explored as an anti-dermatophytic drug.

Chemical constituents from ethanol extract of Polyalthia rumphii branches and their cytotoxicity evaluation

ABSTRACT Twelve known compounds, including eight alkaloids, three lignans and one gossypol derivative, were isolated from the branches of Polyalthia rumphii (Blume ex Hensch.) Merr., Annonaceae. The chemical structures were determined by spectroscopic methods and comparison with literature data. All the isolates were evaluated the cytotoxicity against three human cancer cell lines: Hela, MCF-7 and A549, the results showed that partial of isolates displayed weak cytotoxicities with the IC50 values ranging from 25 to 40 µg/ml.

Effect of cottonseed oil and ferrous sulfate in broiler diets on intestinal morphometry at 42 days of age / Efeito de óleo de semente de algodão e sulfato ferroso em dietas de aves na morfometria intestinal aos 42 dias de idade

The aim of the study was to evaluate the intestinal morphometry of broilers at 42 days of age fed diets containing different levels of cottonseed oil, with and without addition of ferrous sulfate. A total of 560 male Ross chickens were used in a completely randomized design with a 4×2 factorial arrangement and five replications with 14 birds. The cottonseed oil levels tested were 0%, 2%, 4%, and 6%. At 42 days of age, one bird per replicate was collected for morphometric. Analyzed variables were villus height, crypt length and muscle wall thickness. The statistical analysis was performed using SAS, P=0.05. In the duodenum and ileum, there was no significant effect of interaction between the treatments. In the jejunum, only the length of the crypt, the treatment with cottonseed oil 2%, more ferrous sulfate resulted in lower values; however, the same oil content without ferrous sulfate led to a greater length, which was also observed in the treatment with 4% oil plus ferrous sulfate. Inclusion of cottonseed oil in broiler diets at 42 days of age does not impair their intestinal morphology. Supplementation with ferrous sulfate at these levels is unnecessary, since no improvement was observed in the intestinal morphometric.(AU)

O objetivo de estudo foi avaliar a morfometria intestinal de aves aos 42 dias de vida alimentadas com dietas contendo níveis variados de óleo de semente de algodão com e sem adição de sulfato ferroso. Um total de 560 machos de aves Ross foram usadas em estudo randomizado com amostras 4x2 e 5 replicações com 14 aves. Os níveis de óleo de semente de algodão testados foram 0%, 2%, 4%, e 6%. Aos 42 dias de idade, um pássaro por replica foi coletado para morfometria. As variáveis analisadas foram altura de villus, comprimento de cripta, e espessura de parede muscular. A análise estatística foi realizada com SAS, P=0,05. Não houve efeito significativo de interação entre tratamentos no duodeno ou íleo. No jejuno, apenas o comprimento da cripta, no tratamento com óleo de semente de algodão a 2% com mais sulfato ferroso, apresentou valor mais baixo. No entanto, o mesmo valor de óleo sem sulfato ferroso resultou em maior comprimento, resultado percebido também com 4% de óleo. Inclusão de óleo de semente de algodão em dietas de aves aos 42 dias de idade não causa danos à morfologia intestinal. Suplementação com sulfato ferroso não é necessário, já que não houve melhora em morfometria intestinal.

The effect of feed intake containing whole cottonseed on blood parameters of Nellore bulls / Efeito do consumo de rações contendo caroço de algodão sobre parâmetros sanguíneos de bovinos Nelore

The aim of this study was to evaluate the effect of diets containing whole cottonseed (WC) on blood biochemical parameters of bulls. Thirty bulls with 30±6 months and 382.7±28.4kg were kept in feedlot (85 days) and fed the following WC levels: 0, 2.22, 4.44, 6.66, 8.88 and 11.11%. In comparing biochemical indicators from the beginning to the end of the experiment, the control group (CG) and those fed diets containing 2.22, 8.88 and 11.11% of WC had an increase (P<0.05) in serum Ca concentrations (8.34±0.65−9.56±0.92mEqL-1). In relation to Fe (202.79±69.04−300.04±79.88µg/dL), the CG and those treated with 6.66% WC showed an increase (P<0.05) in serum concentrations. As to the Mg (1.92±0.18−2.40±0.27mEqL-1), groups treated with diet containing 2.22, 4.44 and 6.66% of WC had higher (p<0.05) concentrations at the end of the study. Regarding blood lipids, groups with diets containing 2.22; 4.44; 6.66 and 8.88% of WC increased (p<0.05), respectively, in concentrations of HDL, TG, VLDL and COL. The group that received 11.11% of WC increased (P<0.05) in the concentrations of COL and HDL. It can be concluded that diets containing WC, caused no alterations in the concentrations of blood parameters analyzed in this study, with the exception of Mg.(AU)

Objetivou-se avaliar o efeito de dietas contendo caroço de algodão (CA) sobre os parâmetros sanguíneos de bovinos. Trinta touros com 30±6 meses e 382,7±28,4kg foram confinados (85 dias) e receberam dietas com as seguintes proporções de CA: 0; 2,22; 4,44; 6,66; 8,88, e 11,11%. Ao se compararem as concentrações dos indicadores bioquímicos do início com as do final do experimento, foi observado que o grupo controle e aqueles com dietas contendo 2,22; 8,88 e 11,11% de CA apresentaram aumento (P<0,05) na concentração sérica de Ca (8,34±0,65−9,56±0,92 mEqL-1). Já em relação ao Fe (202,79±69,04−300,04±79,88µg/dL), observou-se que apenas o grupo controle e aquele tratado com 6,66% apresentaram aumento (P<0,05) em sua concentração. Quanto ao Mg (1,92±0,18−2,40±0,27 mEqL-1), os grupos tratados com 2,22; 4,44 e 6,66% de CA apresentaram maiores concentrações (P<0,05) ao final do estudo. Com relação aos lipídeos sanguíneos, os grupos com dietas contendo 2,22; 4,44; 6,66 e 8,88% de CA tiveram aumento (P<0,05), respectivamente, nas concentrações de HDL, TG, VLDL e COL. O grupo que recebeu 11,11% de CA teve aumento (P<0,05) nas concentrações de COL e HDL. Conclui-se que as dietas contendo CA não causaram alterações (P>0,05) nas concentrações dos parâmetros sanguíneos analisados, com exceção do Mg.(AU)

Synthesis and Anti-HIV-1 Activity of One Schiff Base Derivative with Glucosamine of Gossypol / 中国药师

Objective: To confirm the structure and preferential conformation of the Schiff base of gossypol with 1, 3, 4, 6-tetra-O-acetyl-β-D-glucosamine and explore its anti-HIV-1 activity.Methods: The Schiff base of gossypol with 11, 3, 4, 6-tetra-O-acetyl-β-D-glucosamine was synthesized and identified by FT-IR, NMR spectroscopy and the PM6 semi-classical calculation.The inhibitory activity of the novel compound against the laboratory-adapted HIV-1IIIB strain was examined using the HIV-1IIIB/TZM-bl indicator cell culture system.Results: The 1H and 13C-NMR signals of the new Schiff base were assigned.The PM6 semi-classical calculation indicated that enamine-enamine tautomeric form of the new Schiff base was more stable,which was stabilized by the intramolecular hydrogen bonds.The anti-HIV-1 test showed that the compound could block the entry of HIV-1IIIB into the target cells.Conclusion: The Schiff base of gossypol with 1, 3, 4, 6-tetra-O-acetyl-β-D-glucosamine exhibits enamine-enamine tautomeric form in solution, which shows potential anti-HIV-1 activity.

Gossypol alleviates isolated heart ischemia/reperfusion injury in rats by interfering with the oxidative stress response and suppressing the JNK/p38 MAPK signal pathway / 安徽医科大学学报

Objective To investigate the protective effect and possible mechanism of Gossypol on isolated myocardial ischemia/reperfusion injury in rats.Methods 40 male Sprague-Dawley rats were randomly divided into 4 groups:Blank group,heart ischemia reperfusion group (MI/R group),high and low-dose Gossypol group (40,20 mg/L).The model of the myocardial ischemia/reperfusion injury were established using the Langendorff method.The hemodynamicsindexes,cardiac enzymes AST and LDH,inflammatory cytokines (NF-κB,ICAM-1,TNF-α and IL-6) were measured.The effect and mechanism of Gossypol on early-stage MI/R of the oxidative stress response and the JNK/p38 MAPK signal pathway were investigated.Results Experimental results showed that Gossypol could significantly improve the functional capacity of the heart,reduce the contents of AST,LDH and inflammatory cytokines in reperfused heart tissue,and increase superoxide dismutase levels to protect the heart.The mechanism of this substance may involve anti-lipid peroxidation and inhibition of p38 kinase phosphorylation and JNK,and reduction of oxidative stress injury and apoptosis damage induced by MI/R.Conclusion This study confirm that Gossypol exerts extensive anti-MI/R effects.Its mechanism may be related to the interfering with the oxidative stress response and suppressing the JNK/p38 MAPK signal pathway.

Amino-functionalized poloxamer 407 with both mucoadhesive and thermosensitive properties: preparation, characterization and application in a vaginal drug delivery system

Lack of mucoadhesive properties is the major drawback to poloxamer 407 (F127)-basedhydrogels for mucosal administration. The objective of the present study was to construct a novel mucoadhesive and thermosensitivehydrogel drug delivery system based on an amino-functionalized poloxamer for vaginal administration. First, amino-functionalized poloxamer 407 (F127-NH) was synthesized and characterized with respect to its micellization behavior and interaction with mucin. Then using acetate gossypol (AG) as model drug, AG-loaded F127-NH-basedhydrogels (NFGs) were evaluated with respect to rheology, drug release,vaginal mucosal adhesion,intravaginal retention and local irritation after vaginal administration to healthy female mice. The results show that F127-NHis capable of forming a thermosensitivehydrogel with sustained drug release properties. An interaction between positively charged F127-NHand negatively charged mucin was revealed by changes in the particle size and zeta potential of mucin particles as well as an increase in the complex modulus of NFG caused by mucin.andfluorescence imaging and quantitative analysis of the amount of AG remaining in mouse vaginal lavage all demonstrated greater intravaginal retention of NFG than that of an unmodified F127-basedhydrogel. In conclusion, amino group functionalization confers valuable mucoadhesive properties on poloxamer 407.

Meat characteristics of cattle fed diets containing whole cottonseed / Características da carne de bovinos alimentados com dietas contendo caroço de algodão

Sensory and instrumental analyses evaluated the meat and fat characteristics of feedlot-finished steers fed a diet containing cottonseed. Global impression, texture, meat color, and fat color were assessed. Thirty Nellore bulls with an average age of 30 ± 6 months and initial body weight of 382.7 ± 28.4kg were kept in feedlot stalls and fed the following cottonseed levels: 0; 2.22 %; 4.44 %; 6.66 %; 8.88 %; 11.11 % of the dietary dry matter. The cottonseed used in this experiment had an average free gossypol content of 4.5g/kg of cottonseed. The overall impression of the samples, assessed by the triangle test for difference, did not differ for more than 62 % of the panelists (P>0.01). The shear strength of roasted meat varied from 6.00 to 6.54kg. According to texture profile analysis (TPA), the hardness, springiness, and chewiness of roast meat ranged from 24.15 to 28.01 N, 0.52 to 0.56, 8.42 to 11.01 N, respectively; of raw meat, 9.51 to 13.86 N, 0.26 to 0.29, and 1.38 to 1.81 N, respectively. The different treatments did not affect meat texture, meat color, or fat color (P>0.05). Meat color, luminosity, and red intensity ranged from 37.71 to 42.85, 20.68 to 25.25, and 6.74 to 8.61, respectively; fat color, luminosity, and yellow intensity ranged from 62.26 to 63.78, 11.13 to 11.62, and 10.53 to 10.86, respectively. Cottonseed intake of up to 1.13kg/animal/day, equivalent to a free gossypol intake of 5.05g/animal/day, in place of soybean meal and ground corn, did not significantly change the global sensory impression, texture, and color of the meat and fat.(AU)

Avaliou-se, por meio de análise sensorial e instrumental, o efeito da adição de caroço de algodão à dieta de bovinos confinados sobre as características da carne e da gordura. Foram avaliadas a impressão global, a textura, a cor da carne e a cor da gordura. Trinta touros da raça Nelore, com médias de idade e peso vivo inicial de 30 ± 6 meses e 382,7 ± 28,4kg, foram confinados e receberam dietas com os seguintes teores de caroço de algodão: 0; 2,22%; 4,44%; 6,66%; 8,88%; 11,11% na matéria seca da dieta. O caroço de algodão utilizado neste experimento apresentou conteúdo médio de 4,5g de gossipol livre/kg de caroço de algodão. A impressão global da carne, avaliada por meio de teste triangular de diferença, mostrou que mais de 62% dos provadores não perceberam diferença significativa (P>0,01) entre as amostras. A textura da carne assada avaliada por meio do teste de força de cisalhamento variou de 6,00 a 6,54kg. Na análise do perfil de textura (TPA) da carne assada, a dureza, a elasticidade e a mastigabilidade variaram, respectivamente, de 24,15 a 28,01N, de 0,52 a 0,56N e de 8,42 a 11,01N. Na TPA da carne crua, variaram, respectivamente, de 9,51 a 13,86N, de 0,26 a 0,29N e de 1,38 a 1,81N. Na avaliação da cor da carne, a luminosidade, a intensidade de vermelho e a intensidade de amarelo da cor da carne variaram, respectivamente, de 37,71 a 42,85, de 20,68 a 25,25 e de 6,74 a 8,61. E para a cor da gordura, variaram, respectivamente, de 62,26 a 63,78, de 11,13 a 11,62 e de 10,53 a 10,86. A textura, a cor da carne e a cor da gordura não apresentaram diferenças significativas (P>0,05) entre os diferentes tratamentos. O consumo de caroço de algodão em até 1,13kg/animal/dia, que resultou no consumo de 5,05g de gossipol livre/animal/dia, em substituição ao farelo de soja e ao grão de milho triturado, não causou alterações significativas nas características da carne quanto à impressão sensorial global, à textura, à cor da carne e à cor da gordura.(AU)

Mechanisms involved in reproductive damage caused by gossypol in rats and protective effects of vitamin E

BACKGROUND: Gossypol is a chemical present in the seeds of cotton plants (Gossypium sp.) that reduces fertility in farm animals. Vitamin E is an antioxidant and may help to protect cells and tissues against the deleterious effects of free radicals. The aim of this study was to evaluate the mechanisms of reproductive toxicity of gossypol in rats and the protective effects of vitamin E. Forty Wistar rats were used, divided into four experimental groups (n = 10): DMSO/ saline + corn oil; DMSO/saline + vitamin E; gossypol + corn oil; and gossypol + vitamin E. RESULTS: Fertility was significantly reduced in male rats treated with gossypol in that a significant decrease in epididy-mal sperm count was observed (P < 0.05) and the number of offspring was significantly reduced in females mated with them (P < 0.05). This dysfunction was prevented by vitamin E. Gossypol caused a significant increase in the activity of the enzymes glutathione peroxidase (P < 0.01) and glutathione reductase (P < 0.01), but vitamin E did not reduce the enzyme activities (P > 0.05). The levels of reduced glutathione and pyridine nucleotides in testis homogen-ate were significantly reduced by gossypol (P < 0.05 and P < 0.01, respectively) and this reduction was accompanied by increased levels of oxidized glutathione (P < 0.05). Vitamin E showed a preventive effect on the changes in the levels of these substances. Gossypol significantly increased the levels of malondialdehyde (P < 0.01), a lipid peroxida-tion indicator, whereas treatment with vitamin E inhibited the action of the gossypol. Vitamin E prevented a decrease in mitochondrial ATP induced by gossypol (P < 0.05). CONCLUSIONS: This study suggests that the reproductive dysfunction caused by gossypol may be related to oxidative stress and mitochondrial bioenergetic damage and that treatment with vitamin E can prevent the infertility caused by the toxin.

Inhibitory effect of gossypol on migration of gastric carcinoma cell lines through Akt/β-catenin passway / 中国药理学通报

Aim To investigate the inhibitory effects of gossypol on migration in gastric carcinoma cell lines and its mechanisms. Methods Gastric carcinoma cells were treated with gossypol at different concentra-tions. The effects of gossypol on cells proliferation were measured using the MTT assay. The migration of gas-tric carcinoma cells was detected by transwell assay. The activation of Akt/β-catenin pathway and the ex-pressions of pathway related proteins ( p-Akt,β-cate-nin, cyclin D1, MMP-2, E-cadherin and vimentin ) were detected by Western blot. Results Gossypol treatment could significantly inhibit the proliferation of gastric carcinoma cells in a dose-dependent manner. Transwell assay showed that the migration ability of gastric carcinoma cells was significantly decreased. The inhibitory effect of gossypol on cells migration was more significant than the effect of gossypol on cells prolifera-tion. Compared with the control group, treatment with gossypol significantly suppressed the expressions of p-Akt,β-catenin, cyclin D1, MMP-2 and vimentin pro-tein, whereas the expression of E-cadherin was signifi-cantly up-regulated in gastric carcinoma cells in a dose-dependent manner. Conclusion These results demonstrate that gossypol represses cell migration of gastric carcinoma cells through the down-regulation of the activity of Akt/β-catenin pathway.

Interaction and Molecular Simulation between Gossypol and Human Serum Albumin / 中国药师

Objective:To explore the interaction between gossypol and human serum albumin ( HSA) . Methods:The interaction of gossypol and HSA under physiological conditions was studied by fluorescence spectroscopy, and the molecular docking software was used to simulate the interaction. Results:The binding constant of gossypol and HSA at 293K and 303K was 2. 390 6 × 105 L·mol-1 and 3. 576 8 × 103 L·mol-1 , respectively. There was one binding site on HSA for gossypol. Hydrogen bond and Van Der Waals inter-actions were involved in the binding process. The binding of gossypol and HAS was closer to tyrosine residue in HSA. The molecular simulation analysis verified the above results. Conclusion: The gossypol-induced fluorescence quenching of HSA belongs to a static quenching procedure.

Isochamaejasmin induces apoptosis in leukemia cells through inhibiting Bcl-2 family proteins / 中国天然药物

The biflavonoid isochamaejasmin is mainly distributed in the root of Stellera chamaejasme L. (Thymelaeaceae) that is used in traditional Chinese medicine (TCM) to treat tumors, tuberculosis, and psoriasis. Herein, isochamaejasmin was found to show similar bioactivity against Bcl-2 family proteins to the reference Bcl-2 ligand (-)-gossypol through 3D similarity search. It selectively bound to Bcl-xl and Mcl-1 with Ki values being 1.93 ± 0.13 μmol·L(-1) and 9.98 ± 0.21 μmol·L(-1), respectively. In addition, isochamaejasmin showed slight growth inhibitory activity against HL-60 with IC50 value being 50.40 ± 1.21 μmol·L(-1) and moderate growth inhibitory activity against K562 cells with IC50 value being 24.51 ± 1.62 μmol·L(-1). Furthermore, isochamaejasmin induced apoptosis of K562 cells by increasing the intracellular expression levels of proteins of the cleavage of caspase-9, caspase-3, and PARP which involved in the Bcl-2-induced apoptosis pathway. These results indicated that isochamaejasmin induces apoptosis in leukemia cells by inhibiting the activity of Bcl-2 family proteins, providing evidence for further studying the underlying anti-cancer mechanism of S. chamaejasme L.

Nanosuspensions study of gossypol: preparation, characterization and in situ single pass perfusion intestinal absorption / 中国药学杂志

OBJECTIVE: To prepare gossypol nanosuspensions and investigate their physicochemical properties and intestinal absorption behavior in rats. METHODS: Gossypol nanosuspensions were prepared by a high pressure homogenization method. Nanosuspensions' characteristics were investigated from particle size distribution, Zeta potential, particle morphology, crystal physical state, storage stability, saturation solubility, and in vitro release. Nanosuspensions' intestinal absorption characteristics were also observed in rats. RESULTS: The nanosuspensions existed as uniform rods. The mean particle size was (217±23) nm, and Zeta potential was -22.7 mV. Gossypol existed in crystalline form. Long-term stability test showed that gossypol nanosuspensions were stable at 4°C for 3 months. The experiment of intestinal absorption showed that the nanosuspensions could be absorbed through passive diffusion with significant variation at different sections. The experiment also showed that the absorption process was not affected by the efflux of P-glyco-protein (drug efflux pump). CONCLUSION: Gossypol nanosuspensions are successfully developed. The solubility in water and the release in vitro of the nanosuspensions are enhanced compared with the bulk drug of gossypol. This study provides a reference for the further pharmaceutical study of gossypol.

Effects of Gossypol Acetic Acid on Protein and mRNA Expressions of hMLH1 Gene in Human Tongue Carcinoma Cell Line Tca8113 / 昆明医科大学学报

Objective The purpose of this study was to investigate the effects of gossypol acetic acid (GAA) on protein and mRNA expressions of hMLH1 gene in human tongue carcinoma cell line Tca8113 in vitro in order to discuss the mechanism of tumor suppression of GAA. Methods (1) Western-blot was used to study the effects of GAA on protein expressions of hMLH1 gene in Tca8113 cell line treated by different concentrations of GAA for 48 h. (2) Real-time fluorescence quantitative PCR (RFQ-PCR) was used to investigate the effects on the mRNA expressions of hMLH1 gene in Tca8113 cell line treated by GAA for 48 h. Results (1) Compared with the control group, the results of Western-blot showed that the protein expression of hMLH1 gene was increased after treatment by GAA for 48 h ( <0.05) . (2) The results of RFQ-PCR indicated that the mRNA expression of hMLH1 gene was increased after GAA treatment for 48 h ( <0.05) . Conclusion GAA could up-regulate protein and mRNA expression of hMLH1 in Tca8113 cell line, which indicated that it may be one of the mechanisms of tumor suppression effect of GAA.

The effects of gossypol acetic acid on DNA methylation level and the mRNA expression of hMLH1 gene in human tongue carcinoma Tca8 1 1 3 cells / 实用口腔医学杂志

Objective:To study the effects of gossypol acetic acid(GAA)on the methylation level and the mRNA expression of hM-LH1 in human tongue cancer Tca8113 cells.Methods:Tca8113 cells were treated by GAA at various doses for 24 h,48 h and 72 h respectively.MTT assay was used to detect the cell proliferation.Nested methylation specific PCR(nMSP)was used to detect methyl-ation level of hMLH1 .Real-time fluorescence quantitative PCR(RFQ-PCR)was applied to investigate the mRNA expression of hM-LH1 gene.Results:GAA inhibited the proliferation of Tca8113 cells dose-and-time dependently,decreased the DNA methylation level of hMLH1(P<0.05)and increased hMLH1 mRNA expression in Tca8113 cells(P<0.05).Conclusion:GAA can suppress proliferation of Tca8113 cells by demethylation of hMLH1 gene and increase of hMLH1 mRNA expression.

Mecanismos da intoxicação do fígado de rato causada pelo gossipol / Mechanisms of the intoxication of rat liver caused by gossypol

O fígado desempenha uma função central no metabolismo devido à sua interposição entre o trato digestivo e a circulação geral do organismo. Ele é também o principal órgão envolvido na biotransformação de substâncias exógenas (xenobióticos), com capacidade de converter compostos hidrofóbicos em hidrossolúveis, mais facilmente eliminados pelo organismo. O gossipol é uma substância fenólica tóxica presente na semente de algodão (Gossypium sp). Com o objetivo de estudar os mecanismos envolvidos na hepatotoxicidade do gossipol avaliou-se os seus efeitos no sistema antioxidante do fígado de ratos no que diz respeito ao estresse oxidativo e aspectos histopatológicos. Foram utilizados ratos machos da linhagem Wistar, separados em dois grupos, sendo que um recebeu óleo de canola (veículo, grupo Controle) e o outro recebeu gossipol na dosagem de 40 mg/kg de peso vivo do animal por 15 dias (grupo Tratado). O tratamento com gossipol promoveu alterações na atividade sérica das enzimas marcadoras de dano hepático e um significativo estresse oxidativo caracterizado pela diminuição nos níveis da glutationa reduzida (GSH) e consequente aumento da glutationa oxidada (GSSG), incluindo, ainda, danos à membrana plasmática e de organelas demonstrados pela peroxidação lipídica. O resultado da avaliação histopatológica demonstrou degeneração dos hepatócitos.

The liver plays a central role in metabolism due to its interposition between the digestive tract and the general circulation of the organism. It is also the main organ involved in biotransformation of exogenous substances (xenobiotics), with ability to convert hydrophobic compounds in water-soluble, more easily eliminated by the body. Gossypol is a toxic phenolic substance present in cotton seed (Gossypium sp.). Aiming to study the mechanisms involved in the hepatotoxicity of gossypol we evaluate its effects on the antioxidant system of rat liver performing an experiment that investigated the oxidative stress and the histopathological alterations. In this study, we used Wistar rats, divided into two groups, one that received canola oil (vehicle, Control group) and another that received gossypol at a dose of 40mg/kg body weight of the animal for 15 days (Treated group). The treatment with gossypol caused alterations in the activity of seric enzymes that indicate hepatic injury and a significant oxidative stress characterized by a decrease of reduced glutathione (GSH) levels and a consequent increase in oxidized glutathione (GSSG), including further damage to the plasma membrane and organelles showed by lipid peroxidation. The result of histopathological evaluation showed degeneration of the hepatocytes.

Biological aspects of Aphis gossypii Glõver, 1877 (Hemiptera: Aphididae) on colored lint cotton cultivars / Aspectos biológicos de Aphis gossypii Glõver, 1877 (Hemiptera: Aphididae) em cultivares de algodoeiro com fibra colorida

This paper outlines the results of a study into the biological aspects of Aphis gossypii Glõver on colored cotton cultivars through the construction of life tables. In addition, we evaluate the influence of density of gossypol glands and trichomes of leaves on the biology of the aphid. Tests were conducted in a climate chamber at 25 ± 2C, with relative humidity (RH) at 70 ± 10%, and a photophase of 12 hours, using the cultivars BRS Rubi, BRS Safira, and BRS Verde. Newly hatched nymphs were individually isolated in petri dishes containing leaf discs of cotton cultivars on a layer of water-agar (1%) of approximately 5 mm thick. An evaluation of the trichomes and gossypol gland densities of the cotton leaf plants was performed under a stereomicroscope, delimited to an area of 1 cm², and a count and identification of those on the surface was made. The feed substrates that were evaluated influenced the nymphal stage of A. gossypii. BRS Verde provided the shortest duration and BRS Safira was the longest during this phase. The cultivar BRS Verde, with the lowest density of trichomes, provided a large number of nymphs and led to a higher net reproductive rate (R0). Given these results, conclusions can be drawn that the colored cotton cultivars influence the duration of nymphal and adult stages of A. gossypii. The cultivar with a high density of trichomes on leaves (BRS Safira) adverse [...]

Este trabalho teve por objetivos estudar aspectos biológicos de Aphis gossypii em cultivares de algodoeiro de fibra colorida, construir tabelas de vida e avaliar a influência da densidade de tricomas e glândulas de gossipol das folhas das plantas na biologia do pulgão. Os ensaios foram conduzidos em câmara climatizada regulada a 25 ± 2C, UR de 70 ± 10% e fotofase de 12 horas, utilizando-se as seguintes cultivares: BRS Rubi, BRS Safira e BRS Verde. Ninfas recém-nascidas foram isoladas individualmente em placas de Petri contendo discos foliares de algodoeiro das cultivares sobre uma lâmina de ágar-água (1%) de aproximadamente 5 mm. A avaliação da densidade de tricomas e de glândulas de gossipol das folhas das plantas foi realizada sob microscópio estereoscópico, delimitando-se uma área de 1 cm² e, em seguida, realizando-se a contagem e identificação dos mesmos nessa superfície. Os substratos alimentares avaliados influenciaram a fase ninfal de A. gossypii, sendo que a BRS Verde proporcionou menor tempo de duração e a BRS Safira, o maior período desta fase. A cultivar BRS Verde com a menor densidade de tricomas proporcionou elevada produção de ninfas e a maior taxa líquida de reprodução (R0). Diante dos resultados, conclui-se que as cultivares de algodoeiro colorido influenciam o tempo de duração da fase ninfal e adulta do pulgão A. gossypii. A cultivar com elevada densidade [...]

Effects of gossypol acetic acid on apoptosis and DNMT1 expression of human tongue carcinoma cell line Tca8113 / 肿瘤

Objective: To investigate the effects of GAA (gossypol acetic acid) on the proliferation, apoptosis and DNMT1 (DNA methyltransferase 1) mRNA expression of human tongue carcinoma cell line Tca8113 in vitro. Methods: The proliferation and apoptosis of Tca8113 cells after treatment with different concentrations of GAA were detected by MTT and flow cytometry, respectively. The expression level of DNMT1 mRNA was examined by real-time fluorescence quantitative-PCR. Results: The proliferative abilities of Tca8113 cells were inhibited after treatment with GAA for 24, 48, and 72 h. The apoptotic rates of Tca8113 cells after treatment with 30 μmol/L GAA for 48 h and 15 μmol/L GAA for 72 h were higher than those of the control cells (without GAA treatment) (P < 0.05). The expression levels of DNMT1 mRNA in Tca8113 cells after treatment with 5, 10, 15 and 20 μmol/L GAA for 48 h were lower than those of the control cells (P < 0.05). Conclusion: GAA can inhibit the proliferation and induce the apoptosis of human tongue carcinoma cell line Tca8113 with a decreased expression level of DNMT1 mRNA. Copyright © 2013 by TUMOR.

Gossypol acetic acid induces apoptosis in RAW264.7 cells via a caspase-dependent mitochondrial signaling pathway

To investigate the effects of gossypol acetic acid (GA) on proliferation and apoptosis of the macrophage cell line RAW264.7 and further understand the possible underlying mechanism responsible for GA-induced cell apoptosis, RAW264.7 cells were treated with GA (25~35 micromol/L) for 24 h and the cytotoxicity was determined by MTT assay, while apoptotic cells were identified by TUNEL assay, acridine orange/ethidium bromide staining and flow cytometry. Moreover, mitochondrial membrane potential (DeltaPsi(m)) with Rhodamine 123 and reactive oxygen species (ROS) with DCFH-DA were analyzed by fluorescence spectrofluorometry. In addition, the expression of caspase-3 and caspase-9 was assessed by Western Blot assay. Finally, the GA-induced cell apoptosis was evaluated by flow cytometry in the present of caspase inhibitors Z-VAD-FMK and Ac-LEHD-FMK, respectively. GA significantly inhibited the proliferation of RAW264.7 cells in a dose-dependent manner, and caused obvious cell apoptosis and a loss of DeltaPsi(m) in RAW264.7 cells. Moreover, the ROS production in cells was elevated, and the levels of activated caspase-3 and caspase-9 were up-regulated in a dose-dependent manner. Notably, GA-induced cell apoptosis was markedly inhibited by caspase inhibitors. These results suggest that GA-induced RAW264.7 cell apoptosis may be mediated via a caspase-dependent mitochondrial signaling pathway.