Protective effects of Angelica Sinensis polysacchairides on the testis of aging model mice induced by D-galactose / 解剖学报

Objective: To investigate the protective effects of Angelica Sinensis polysacchairides (ASP) on the testis of aging mice induced by D-galactose(D-Gal) and its mechanism. Methods: Forty male C57BL/6 J mice were randomly divided into the normal,ASP normal,aging model and ASP aging model groups,with 10 mice in each group.The aging model group mice were subeutaneously injected with D-Gal(120 mg/kg, qd Ã- 42); the normal group mice were subcutaneously injected with saline of the same dose at the same time; the ASP normal group mice were subeutaneously injected with the same amount of saline,qdx 15,and following intraperitoneally injected with ASP(140 mg/kg,qdÃ-27);the ASP aging model group were the same as the aging model group, from the 16th day of establishment of the aging model group on, the mice were subcutaneously injected with the same dose and the same time of ASP.The 2nd day after model establishment, peripheral blood was collected from the inner canthus vein and the content of serum testosterone was measured;the testes were sampled and testicular index determined; paraffin sections and HE staining of the testes were prepared, the testis histopathology was observed; frozen sections of the testes were prepared, the aging of testicular cells was detected by senescence associated-|3-galactosidase (SA-p-Gal) staining; the tissue homogenate of testes was prepared, the activity of superoxide dismutase (SOD) was evaluated by enzymatic method, the total antioxidant capacity (T-AOC) was detected by 2,2' -amino-di(3-ethyl-benzothiazoline sulphonic acid-6) ammonium salt(ABTS),the content of malondialdehyde (M DA) was measured by thiobarbituric acid method; the expression of aging-related proteins P53 and P21 was detected by Western blotting. Results There is no significant difference in the wet weight of testis and the index of testicular organs in the groups,but the testicular tissue structure of the aging model group was obviously damaged.The layers of the spermatogenic cells in seminiferous epithelium, the testicular interstitial cells and the serum testosterone level significantly decreased; the positive cells detected by SA-Î2-Gal stain markedly increased; the activity of superoxide dismutase (SOD) and the total antioxidant capacity(T-AOC) obviously decreased,while the content of malondialdehyde(MDA) increased significantly;and the expression of P53 and P21 was evidently up-regulated.Compared the ASP aging model group with the aging model group, the testicular tissue structure was not obviously damaged.The number of the spermatogenic cells,the testicular interstitial cells and the serum testosterone level decreased less evidently, the SA-Î2-Gal stain positive cells decreased significantly; the activity of superoxide dismutase (SOD) and the total antioxidant capacity (T-AOC) increased significantly, while the content of malondialdehyde(MDA) decreased significantly; and the expression of P53 and P21 was remarkably down-regulated. Conclusion: ASP can antagonize testis aging induced by D-galactose in mice. Inhibition of oxidative stress damage and the expression of senescence-associated genes may be the underlying mechanism.

Western diet induces endogen oxidative deoxyribonucleic acid damage and infl ammation in Wistar rats / Dieta ocidental induz endógeno dano oxidativo do ácido desoxirribonucleico e infl amação em ratos Wistar

ABSTRACT Objective Nutritional diseases such as metabolic syndrome, cardiovascular disorder, chronic inflammation or even cancer are observed in people who sustain their lifestyle by Western diet due to high calorie intake. The origin of these diseases are the degraded deoxyribonucleic acid structure. In this study, we investigated whether Western diet produced endogenous oxidative deoxyribonucleic acid damage, apoptosis or inflammation. Methods Twenty-eight male Wistar rats, aged 10-12 weeks, were divided into four groups. The rats in control group received the standard diet and the remaining rats were given one of the following three diets for four weeks: a high-fat diet containing 35% fat, a high-sucrose diet containing 69% sucrose and Western diet comprising both two types of diets. After treatment the serum 8-hydroxy-2-deoxyguanosine, poly (adenosine diphosphate ribose) polymerase-1, chitinase-3-like protein 1, soluble urokinase-type plasminogen activator receptor, Fas ligand and cytochrome c levels were measured. Results It was observed no changes in the serum soluble urokinase-type plasminogen activator receptor, Fas ligand and cytochrome c levels whereas a statistically significant increase in the serum 8-hydroxy-2-deoxyguanosine, poly (adenosine diphosphate ribose) polymerase-1 and chitinase-3-like protein 1 levels were found only in rats that were given Western diet. Conclusion The findings show that Western diet produced endogenous oxidative deoxyribonucleic acid damage, which then increased serum poly (adenosine diphosphate ribose) polymerase-1 levels, eventually leading to inflammation.

RESUMO Objetivo Doenças nutricionais, como síndrome metabólica, distúrbios cardiovasculares, inflamação crônica ou mesmo câncer, são observadas em pessoas que sustentam seu estilo de vida na dieta ocidental, caracterizada pela alta ingestão de calorias. Dado que a origem dessas doenças é a estrutura degradada do ácido desoxirribonucleico, o presente estudo investigou se a dieta ocidental produzia dano oxidativo endógeno ao ácido desoxirribonucleico, apoptose ou inflamação. Métodos Foram utilizados 28 ratos Wistar machos, com idade entre 10-12 semanas, divididos em quatro grupos. Os ratos do grupo controle receberam a dieta padrão, ao passo que os ratos restantes receberam uma das três dietas seguintes por quatro semanas: uma dieta rica em gordura contendo 35% de gordura; uma dieta rica em sacarose contendo 69% de sacarose; e dieta ocidental compreendendo os dois tipos de dietas. Após o tratamento soro 8-hidroxi-2-desoxiguanosina, poli (adenosina difosfato ribose) polimerase-1, quitinase-3-like proteína 1, uroquinase solúvel tipo de receptor ativador de plasminogênio, os níveis do ligante Fas e do citocromo c foram medidos. Resultados Não foram observadas alterações nos níveis séricos de uroquinase solúvel tipo de receptor ativador de plasminogênio, ligante Fas e citocromo c, enquanto um aumento estatisticamente significativo nos níveis séricos de 8-hidroxi-2-desoxiguanosina, poli (adenosina difosfato ribose) polimerase-1 e quitinase-3-like proteína 1 foi encontrado apenas em ratos que receberam dieta ocidental. Conclusão Os resultados mostram que a dieta ocidental produziu danos no ácido desoxirribonucleico oxidativo endógeno, o que aumentou os níveis séricos de poli (adenosina difosfato ribose) polimerase-1, levando à inflamação.

Rg1 alleviates the damage in ALS model through regulation of miR153/Nrf2/HO-1 / 药学学报

The impact of Rg1 in the disease progress and pathology of amyotrophic lateral sclerosis (ALS) was investigated in mouse model (SOD1 G93A). Body weight and survival rate were monitored to check the course of disease. Rotarod test was used to evaluate the coordination of muscle movement. Toluidine blue staining and immunofluorescence were used to check the effect of Rg1 on motor neuron and microglia. The expression of oxidative stress related protein Nrf2 and the miRNA were tested to investigate the mechanism of Rg1. We found that 20 mg·kg-1·d-1 Rg1 significantly postponed the disease onset and process, improved the motor syndrome, reduced the loss of motor neuron and inhibited the activation of microglia cells. Rg1 inhibited the aggregation of miR-153 in the spinal cord of ALS mice, which relieved the inhibition of Nrf2 and contributed to its up-regulation in the activation of HO-1 anti-oxidative signal pathway. Our study confirmed that Rg1 could protect ALS mice from oxidative damage through the up-regulation of miR-153/Nrf2/HO-1, which provides a theoretical foundation for Rg1 application to the ALS treatment.

Moderate Dose of Trolox Preventing the Deleterious Effects of Wi-Fi Radiation on Spermatozoathrough Reduction of Oxidative Stress Damage / 中华医学杂志(英文版)

<p><b>Background</b>The worsening of semen quality, due to the application of Wi-Fi, can be ameliorated by Vitamin E. This study aimed to demonstrate whether a moderate dose of trolox, a new Vitamin E, inhibits oxidative damage on sperms in vitro after exposure to Wi-Fi radiation.</p><p><b>Methods</b>Each of the twenty qualified semen, gathered from June to October 2014 in eugenics clinic, was separated into four aliquots, including sham, Wi-Fi-exposed, Wi-Fi plus 5 mmol/L trolox, and Wi-Fi plus 10 mmol/L trolox groups. At 0 min, all baseline parameters of the 20 samples were measured in sequence. Reactive oxygen species, glutathione, and superoxide dismutase were evaluated in the four aliquots at 45 and 90 min, as were sperm DNA fragments, sperm mitochondrial potential, relative amplification of sperm mitochondrial DNA, sperm vitality, and progressive and immotility sperm. The parameters were analyzed by one-way analysis of variance and Tukey's posttest.</p><p><b>Results</b>Among Wi-Fi plus 5 mmol/L trolox, Wi-Fi-exposed and Wi-Fi plus 10 mmol/L trolox groups, reactive oxygen species levels (45 min: 3.80 ± 0.41 RLU·10·mlvs. 7.50 ± 0.35 RLU·10·mlvs. 6.70 ± 0.47 RLU·10·ml, P < 0.001; 90 min: 5.40 ± 0.21 RLU·10·mlvs. 10.10 ± 0.31 RLU·10·mlvs. 7.00 ± 0.42 RLU·10·ml, P < 0.001, respectively), percentages of tail DNA (45 min: 16.8 ± 2.0% vs. 31.9 ± 2.5% vs. 61.3 ± 1.6%, P < 0.001; 90 min: 19.7 ± 1.5% vs. 73.7 ± 1.3% vs. 73.1 ± 1.1%, P < 0.001, respectively), 8-hydroxy-2'-deoxyguanosine (45 min: 51.89 ± 1.46 pg/ml vs. 104.89 ± 2.19 pg/ml vs. 106.11 ± 1.81 pg/ml , P = 0.012; 90 min: 79.96 ± 1.73 pg/ml vs. 141.73 ± 2.90 pg/ml vs. 139.06 ± 2.79 pg/ml; P < 0.001), and percentages of immotility sperm (45 min: 27.7 ± 2.7% vs. 41.7 ± 2.2% vs. 41.7 ± 2.5%; 90 min: 29.9 ± 3.3% vs. 58.9 ± 4.0% vs. 63.1 ± 4.0%; all P < 0.001) were lowest, and glutathione peroxidase (45 min: 60.50 ± 1.54 U/ml vs. 37.09 ± 1.77 U/ml vs. 28.18 ± 1.06 U/ml; 90 min: 44.61 ± 1.23 U/ml vs. 16.86 ± 0.93 U/ml vs. 29.94 ± 1.56 U/ml; all P < 0.001), percentages of head DNA (45 min: 83.2 ± 2.0% vs. 68.2 ± 2.5% vs. 38.8 ± 1.6%; 90 min: 80.3 ± 1.5% vs. 26.3 ± 1.3% vs. 26.9 ± 1.1%; all P < 0.001), percentages of sperm vitality (45 min: 89.5 ± 1.6% vs. 70.7 ± 3.1% vs. 57.7 ± 2.4%; 90 min: 80.8 ± 2.2% vs. 40.4 ± 4.0% vs. 34.7 ± 3.9%; all P < 0.001), and progressive sperm (45 min: 69.3 ± 2.7% vs. 55.8 ± 2.2% vs. 55.4 ± 2.5%; 90 min: 67.2 ± 3.3% vs. 38.2 ± 4.0% vs. 33.9 ± 4.0%; all P < 0.001) were highest in Wi-Fi plus 5 mmol/L trolox group at 45 and 90 min, respectively. Other parameters were not affected, while the sham group maintained the baseline.</p><p><b>Conclusion</b>This study found that 5 mmol/L trolox protected the Wi-Fi-exposed semen in vitro from the damage of electromagnetic radiation-induced oxidative stress.</p>

Chemical constituents from petroleum ether fraction of Swertia chirayita and their activities in vitro / 中国中药杂志

The present work is to study the chemical constituents from petroleum ether fraction of Tibetan medicine Swertia chirayita by column chromatography and recrystallization. The structures were identified by physical and chemical properties and spectral data as swerchirin (1), decussatin (2), 1,8-dihydroxy-3,5,7-trimethoxyxanthone (3), 1-hydroxy-3,5,7,8-tetramethoxyxanthone (4), bellidifolin (5), 1-hydroxy-3, 7-dimethoxyxanthone (6), methylswertianin (7), 1-hydroxy-3,5-dimethoxyxanthone (8), erythrodiol (9), oleanolic acid (10), gnetiolactone (11), scopoletin (12), sinapaldehyde (13), syringaldehyde (14), and β-sitosterol (15). Compounds 3, 4, 9, 11-14 were isolated from S. chirayita for the first time. Compounds 9 and 12 were firstly isolated from the genus Swertia. The cytotoxic activities of compounds 1, 2, 5, 7 and 8 against human pancreatic cancer cell lines SW1990 and BxPC-3,and the protective effects of these compounds against hydrogen peroxide (H2O2)-induced oxidative stress in human endothelium-derived EA.hy926 were investigated in vitro. The results showed no obvious effect at the high concentration of 50 μmol•L⁻¹.

Protective effects of penehyclidine hydrochloride on A549 cells against oxidative injury / 天津医药

Objective To explore the effects of penehyclidine hydrochloride (PHC) on lung epithelial type Ⅱcells against oxidative stress damage. Methods A549 cells treated with H2O2 were used as oxidative stress damage cell model. A549 cells were divided into 3 groups: control group (C group), H2O2 treated group (H group) and PHC treated group (P group). The viability of A549 cells was measured by MTT assay. The apoptotic rate was measured by TUNEL assay. The lev?els of malonicdialed (MDA), reactive oxygen species (SOD), reduced glutathione hormone (GSH) and nicotinamide adenine dinucleotide phosphate (NADPH) of cells were detected by biochemistry colorimetry. Results Compared with group C, the viability of A549 and the contents of SOD, GSH and NADPH were significantly decreased in group H, while MDA content and apoptotic rate were increased (P<0.05). Compared with group H, the viability of A549, the contents of SOD, GSH and NADPH were significantly increased in group P, while MDA content and apoptotic rate were reduced (P<0.05). Conclu?sion Penehyclidine hydrochloride shows protective effects on A549 cells through reducing the oxidative damage induced by H2O2.

Antioxidant effect of mogrosides against oxidative stress induced by palmitic acid in mouse insulinoma NIT-1 cells

Excessive oxidative stress in pancreatic β cells, caused by glucose and fatty acids, is associated with the pathogenesis of type 2 diabetes. Mogrosides have shown antioxidant and antidiabetic activities in animal models of diabetes, but the underlying mechanisms remain unclear. This study evaluated the antioxidant effect of mogrosides on insulinoma cells under oxidative stress caused by palmitic acid, and investigated the underlying molecular mechanisms. Mouse insulinoma NIT-1 cells were cultured in medium containing 0.75 mM palmitic acid, mimicking oxidative stress. The effects of 1 mM mogrosides were determined with the dichlorodihydrofluorescein diacetate assay for intracellular reactive oxygen species (ROS) and FITC-Annexin V/PI assay for cell apoptosis. Expression of glucose transporter-2 (GLUT2) and pyruvate kinase was determined by semi-quantitative reverse-transcription polymerase chain reaction. Palmitic acid significantly increased intracellular ROS concentration 2-fold (P<0.05), and decreased expression of GLUT2 (by 60%, P<0.05) and pyruvate kinase (by 80%, P<0.05) mRNAs in NIT-1 cells. Compared with palmitic acid, co-treatment with 1 mM mogrosides for 48 h significantly reduced intracellular ROS concentration and restored mRNA expression levels of GLUT2 and pyruvate kinase. However, mogrosides did not reverse palmitic acid-induced apoptosis in NIT-1 cells. Our results indicate that mogrosides might exert their antioxidant effect by reducing intracellular ROS and regulating expression of genes involved in glucose metabolism. Further research is needed to achieve a better understanding of the signaling pathway involved in the antioxidant effect of mogrosides.

Ulinastatin attenuates paraquat-induced myocardial injury / 中华急诊医学杂志

Objective To investigate the effect of ulinastatin (UTI) on endogenous antioxidant systems in the process of myocardial oxidative stress injury induced by paraquat (PQ),so as to elucidate UTI protecting cadiocytes against PQ-induced injury.Methods Thirty Japan white rabbits were randomly (random number) divided into A,a sham control group; B,PQ intoxication model group and C,D,E,three UTI groups as per different dosages of UTI given.PQ (30 mg/kg) was administered by intraperitoneal route to the rabbits of PQ intoxication model and rabbits of three UTI groups.UTI in doses of 15 000,30 000,50 000 U/kg was administered intravenously every day to the rabbits of three UTI groups respectively for 7 days after PQ intoxication.After the rabbits were sacrificed with 10％ chloral hydrate,left ventricles of rabbits were taken.Histomorphological changes of myocardium were observed by using HE staining.Hydroxyproline was measured by alkaline hydrolysis method,myocardial oxidative stress evaluated by 4-hydroxy-2-nonenal (4-HNE) immunohistochemistry staining,transcription levels of transforming growth factor-β1 (TGF-β1),adiponectin,peroxisome proliferator-activated receptor α (PPAR-α),AMP-activated protein kinase-β1 (AMPK-β1),uncoupling protein-1 (UCP-1) were assayed with reverse transcriptionpolymerase chain reaction (RT-PCR).The differences between two groups were analyzed by t test ; for multiple comparisons,data were analyzed by One-Way ANOVA followed by Dunnett' s post hoc test; for two and multiple ranked data comparisons,Mann-Whitney U test and Kruskal-Wallis test were used,and Spearman rank correlation analysis was used to investigate the correlation between UTI dose and transcription levels of endogenous antioxidant.Results Compared with group A,left ventricular cadiocytes in group B became swelled and disarranged,concentration of tissue HYP was elevated to (2.37 ± 0.49),P =0.001;scores of 4-HNE and TGF-β1 mRNA expressions were all upregulated (both P =0.001).Compared with group B,disordered myocytes in UTI treated groups tended towards normal,4-HNE scores in group D and group E descended respectively to (1.83 ± 0.53) and (1.70 ± 0.47),both P =0.001,HYP concentration reduced to (3.51 ±0.39) μg/mg and (3.29 ±0.37) μg/mg (P =0.002; P =0.001),and expressions of TGF-β1 mRNA in these two groups were decreased (both P =0.001).Transcription levels of four endogenuous antioxidant components,adiponectin,PPAR-α,AMPK-β1 and UCP-1,were all augmented in the group E with high dose UTI.The mRNA expressions of these four components positively correlated with the dose of UTI in the myocardium of PQ intoxicated rabbits.Conclusions UTI could protect myocardia against PQ-induced injury by increasing endogenous antioxidant systems.