Cigarette Smoke Induces Gefitinib Resistance in NSCLC Cells via ROS/Sirt3/SOD2 Pathway / 中国肺癌杂志

BACKGROUND@#Epidermal growth factor receptor (EGFR) gene mutations are the most common driver mutations in non-small cell lung cancer (NSCLC). To prolong the survival of the patients, EGFR tyrosine kinase inhibitors (TKIs) resistance in NSCLC is a major challenge that needs to be addressed urgently, and this study focuses on investigating the mechanism of cigarette smoke (CS) induced Gefitinib resistance in NSCLC.@*METHODS@#PC-9 and A549 cells were cultured in vitro and treated with 1 µmol/L Gefitinib for 4 h and 10% cigarette smoke extract (CSE) for 48 h. Western blot was used to detect Sirtuin 3 (Sirt3) and superoxide dismutase 2 (SOD2) protein expressions; DCFH-DA probe was used to detect intracellular reactive oxygen species (ROS); CCK-8 kit was used to detect cell activity, and EdU was used to detect cell proliferation ability. Sirt3 overexpression plasmid (OV-Sirt3) was transfected in PC-9 and A549 cells and treated with 1 µmol/L Gefitinib for 4 h and 10% CSE for 48 h after N-acetylcysteine (NAC) action. The expressions of Sirt3 and SOD2 were detected by Western blot; the ROS level in the cells was detected by DCFH-DA probe, and the cell activity was detected by CCK-8.@*RESULTS@#CSE induced an increase in the 50% inhibitory concentration (IC50) of both PC-9 and A549 cells to Gefitinib (P<0.01) and enhanced the proliferation of PC-9 and A549 cells, suggesting that CS induced Gefitinib resistance in NSCLC. ROS was involved in CSE-induced Gefitinib resistance (P<0.05). CSE induced low expressions of Sirt3 and SOD2 (P<0.01), and Sirt3/SOD2 was associated with poor prognosis in lung cancer patients (P<0.05). OV-Sirt3 in PC-9 and A549 cells reversed CSE-induced Gefitinib resistance (P<0.05) and significantly reduced ROS production. NAC reversed CSE-induced Gefitinib resistance in PC-9 and A549 cells (P<0.05).@*CONCLUSIONS@#The ROS/Sirt3/SOD2 pathway is involved in CS-induced Gefitinib resistance in NSCLC.

Effect of High-Fat Diet on SOD2, GPx, NeuN and BDNF Expression on Frontal Lobe of Obese Rats

@#Introduction: Obesity has been demonstrated to induce oxidative stress and inflammation processes that lead to senescence in brain cells. Obesity-induced cellular senescence in the brain is still widely investigated. This study aimed to investigate the expression of antioxidant and neuronal markers in the frontal lobes of obese rats. Methods: Eighteen adult rat Sprague Dawley divided into three groups: Control (SO), Obese-2 (DIO2), and Obese-4 (DIO4) were observed. Control rats were fed with a standard diet AIN 76A for two month. In contrast, DIO2 and DIO4 rats were fed with a high-fat diet daily for two and four months, respectively. After being sacrificed, the rats’ brains were dissected out then the frontal lobes were used for RNA extraction. Reverse transcriptase PCR of SOD2, GPx, BDNF, NeuN and beta-actin was performed to investigate the relative expression of the antioxidant and neuronal markers. Results: DIO2 and DIO4 groups had significantly increased body Weights, blood glucose level and triglyceride level after being fed with a high-fat diet for two and four months, respectively. The DIO4 group had the significantly lowest mRNA expressions of SOD2, GPx, BDNF and NeuN. Conclusion: Decreased antioxidant and neuronal markers in the rats frontal lobes were observed as the chronic effect of obesity.

Ginsenoside Rb1 Attenuates Human Umbilical Vein Endothelial Cells Senescence Induced by High Glucose through Sirt3/SOD2 Pathway / 中山大学学报(医学科学版)

@#Abstract】 【Objective】To investigate the effect and mechanism of ginsenoside Rb1 attenuating human umbilical vein endothelial cells(HUVEC) senescence induced by high glucose through Sirt3/SOD2 pathway.【Methods】The senescence of HUVEC induced by high glucose(40 mmol/L)was assessed by senescence-associated β-galactosidase(SA-β-Gal)staining,and the expression of plasminogen activator inhibitor 1(PAI-1)and P16. Annexin V-FITC/PI was performed to measure apoptotic effect. The expression of sirtuins 3(Sirt3)and superoxide dismutase 2(SOD2)was detected by western blot. Meanwhile,the level of intracellular malondialdehyde(MDA)and the activity of SOD2 were measured.【Results】Treatment of HUVEC with high glucose for 24 hours induced premature senescence instead of apoptosis,as indicated by a larger proportion of the cells stained with SA-β-Gal and the up-regulated expression of PAI-1 and P16. Pretreatment of HUVEC with ginsenoside Rb1(40 μmol/L)could reverse endothelial cell senescence,as indicated by the reduced SA-β-Gal positive cells and the down-regulated expression of PAI-1 and P16. Furthermore,ginsenoside Rb1 pretreatment upregulated the protein expression of Sirt3 and SOD2,and eventually increased the activity of SOD2 and decreased the level of MDA.【Conclusion】Ginsenoside Rb1 could antagonize high glucose-induced premature senescence of HUVEC via Sirt3/SOD2 signaling pathway.

Effect of Le ’ermai Capsule on Protein Expression of HO- 1，SOD1，SOD2 and Nrf 2 in the Cerebral Cortex of Cerebral Ischemia/Reperfusion Injury Rats / 中国药房

OBJECTIVE： To study the effects and mechanism of Le’ermai capsule on oxidative stress injury in the cerebral cortex of cerebral ischemia-reperfusion injury rats. METHODS： Totally 65 rats were randomly divided into sham operation group （normal saline）， model group （normal saline）， positive control group （Naoxintong capsule， 3.40 g/kg）， Le’ermai capsule high-dose and low-dose groups （0.37， 0.18 g/kg）， with 13 rats in each group. Except that sham operation group received sham operation， and middle cerebral artery occlusion/reperfusion （MCAO/R） model was induced by suture-occluded method. 24 h after modeling， they were given relevant medicine intragastrically， once a day， for consecutive 7 d. 2 h after last administration， cerebral ischemic area was determined by TTC staining. The pathological changes of cerebral tissue were observed by HE staining. The protein expressions of HO-1， SOD1， SOD2 and Nrf2 in cerebral cortex were detected by Western blot assay. RESULTS： Compared with sham operation group， the area of cerebral ischemic was increased significantly in model group（P＜0.05）， interstitial edema was serious， inflammatory cell infiltration increased significantly； and the protein expression levels of HO-1 and SOD1 in cortex tissue were significantly increased （P＜0.05）， and the protein expression levels of SOD2 and Nrf2 were significantly decreased （P＜0.05）. Compared with model group， the area of cerebral ischemic area of Le’ermai capsule high-dose and low-dose groups were significantly decreased （P＜0.05）. The interstitial edema， inflammatory cell infiltration and microcytes proliferation were decreased. The protein expression levels of HO-1， SOD1， SOD2 and Nrf2 in the cerebral cortex tissue were significantly increased （P＜0.05）. CONCLUSIONS：Le’ermai capsule can improve cerebral ischemia/reperfusion injury to certain extent， and which may be associated with up-regulating the protein expression of antioxidant factors HO-1， SOD1， SOD2 and Nrf2 in the cortex.

Moderate-intensity exercise allows enhanced protection against oxidative stress-induced cardiac dysfunction in spontaneously hypertensive rats

The progression of myocardial injury secondary to hypertension is a complex process related to a series of physiological and molecular factors including oxidative stress. This study aimed to investigate whether moderate-intensity exercise (MIE) could improve cardiac function and oxidative stress in spontaneously hypertensive rats (SHRs). Eight-week-old male SHRs and age-matched male Wistar-Kyoto rats were randomly assigned to exercise training (treadmill running at a speed of 20 m/min for 1 h continuously) or kept sedentary for 16 weeks. Cardiac function was monitored by polygraph; cardiac mitochondrial structure was observed by scanning electron microscope; tissue free radical production was measured using dihydroethidium staining. Expression levels of SIRT3 and SOD2 protein were measured by western blot, and cardiac antioxidants were assessed by assay kits. MIE improved the cardiac function of SHRs by decreasing left ventricular systolic pressure (LVSP), and first derivation of LVP (+LVdP/dtmax and −LVdP/dtmax). In addition, exercise-induced beneficial effects in SHRs were mediated by decreasing damage to myocardial mitochondrial morphology, decreasing production of reactive oxygen species, increasing glutathione level, decreasing oxidized glutathione level, increasing expression of SIRT3/SOD2, and increasing activity of superoxide dismutase. Exercise training in SHRs improved cardiac function by inhibiting hypertension-induced myocardial mitochondrial damage and attenuating oxidative stresses, offering new insights into prevention and treatment of hypertension.

Effect of Cimetidine on the Histopathology and the Expression of SOD2 and GPX1 in Low-dose Accumulative Irradiated Beagle Dogs Spleen / 中国药学杂志

OBJECTIVE: To study the effect of cimetidine on the histopathology and the expression of SOD2, GPX1 gene and protein in low-dose accumulative irradiated Beagle dogs spleen and to investigate the mechanism of cimetidine′s protective effect on low-dose accumulative irradiation. METHODS: Twenty-four male Beagle dogs were divided into six groups: normal control group,model group,positive drug control group and cimetidine groups at the dose of 5.33, 10.67, 21.33 mg•kg-1 respectively. The dogs were irradiated with 60Co-γ-ray at 0.040 8 mGy•min-1 rate for 23 d. Cimitidine was administered intragastrically during irradiation, once a day. Microscope was adopted to observe the pathologic change of spleen and the expression levels of SOD2,GPX1 gene and protein in the spleen tissue of dogs were detected by qRT-PCR and immunohistochemistry techniques. RESULTS: Compared with the model group,cimetidine ameliorated the pathological changes and ultrastructure lesions in the spleen of dogs. Also compared with the model group, the levels of SOD2 and GPX1 protein in cimetidine groups at the dose of 5.33, 10.67, 21.33 mg•kg-1 were higher(P<0.05) and the levels of SOD2 and GPX1 mRNA of the three cimetidine groups were increased significantly(P<0.01). CONCLUSION: Cimetidine can reduce the histological damage. Cimetidine can upregulate the expressions of SOD2 and GPX1 gene and protein and it has good protective effect on the antioxidant system injury.

Protective effects of silymarin on methotrexate-induced damages in rat testes

Abstract The present study aimed to investigate the protective effects of silymarin (SMN), an antioxidant, on methotrexate (MTX)-induced damage in rat testes. Thirty-two Wistar albino rats were divided into four groups (n = 8): control, MTX (20 mg/kg, i.p. on days 1 and 5), SMN (200 mg/kg, orally), and MTX + SMN (20 mg/kg, i.p. on days 1 and 5 and SMN 200 mg/kg orally) groups. At the end of the 6-week trial period, histopathological, immunohistochemical, biochemical, and spermatological analyses were performed on testes tissues. Histopathologically, MTX-induced damage, including depletion of germ cell and loos of spermatozoa, was significantly improved with SMN treatment. Immunohistochemically, the immunoreactivity of glutathione peroxidase 1 (GPx1) and manganese superoxide dismutase 2 (SOD2) were detected more intensely in the MTX + SMN group than in the MTX group. Biochemical examinations revealed that SMN supplementation decreased the lipid peroxidation and increased enzymatic antioxidants in the SMN-treated rats. Spermatologically, significant differences were found in the density, motility, dead-to-live sperm ratio, and abnormal sperm rate in the MTX + SMN group compared to the MTX group. In conclusion, SMN seems to have protective effects as an antioxidant against MTX-induced damage in rat testes.

Protective effect of Lactobacillus casei Zhang against Acetaminophen-induced acute liver injury in mice / 中国免疫学杂志

Objective:To investigate the protective effects of Lactobacillus casei Zhang ( Lcz) on mice with Acetaminophen (APAP)-induced acute liver injury.Methods:Mice were randomly divided into five groups:Control group (ctrl),APAP-induced liver injury group ( APAP ) , N-Acetylcysteine positive control group ( NAC ) , Lcz preventive group ( Lcz/APAP ) and Lcz control group (Lcz).Mice were orally administrated with Lcz (1×109 CFU/ml) for 30 continuous days prior to APAP exposure.Positive control group were intraperitoneally injected with 150 mg/kg NAC 1 h before APAP challenge.Mice in the APAP,NAC and Lcz/APAP group were intraperitoneally injected with 300 mg/kg APAP.Blood and liver samples were collected 18 h post APAP challenge.The expression of HO-1,SOD2,Bcl-2 and TLR4 were detected by Western blot.Results: Lcz could significantly inhibit APAP-induced increase in ALT and AST levels.Furthermore,pretreatment with Lcz increased the hepatic expression of HO-1,SOD2 and Bcl-2,while suppressing the expression of Toll-like receptor 4 (TLR4).Conclusion:Lcz can protect against APAP-induced acute liver injury in mice.The hepato-protective effects of Lcz are associated with its anti-oxidative and anti-inflammatory capacity.

Association of manganese superoxide dismutase gene polymorphism (Ala-9Val) and breast cancer in males and females / Associação do polimorfismo do gene da superóxido dismutase dependente de manganês (Ala-9Val) em homens e mulheres com câncer de mama

INTRODUCTION: One of the several metabolic pathways involved in breast carcinogenesis is the human polymorphism in the mitochondrial targeting sequence Ala-9Val of the manganese superoxide dismutase (MnSOD) gene, which has been previously associated with increased risk of breast cancer in females. Since there is no previous report on this polymorphism in male breast cancer, the objective of this study is to analyze MnSOD polymorphism in a population of males and females with breast cancer from the southernmost state of Brazil, compared to healthy controls. METHODS: A case-control study of one hundred patients affected by breast cancer (11 men and 89 women) and 370 healthy age-adjusted database controls was performed. DNA was extracted from paraffin-embedded tumoral tissue. MnSOD polymorphism was determined by PCR-RFLP techniques using restriction enzyme Hae III. Chi-square test was used to compare MnSOD frequency distribution. RESULTS: MnSOD genotypic frequencies in all patients with breast cancer were AA = 15 percent; AV = 56 percent; VV = 29 percent and controls AA = 6.5 percent; AV = 68.1 percent and VV = 25.4 percent. Both male and female patients with breast cancer presented significantly higher AA frequencies compared to controls (p = 0.035), suggesting strong association of this genotype with breast cancer. A 2.15-fold (95 percent confidence interval [CI] 1.393-4.541) risk of breast cancer was found among individuals carrying the MnSOD AA allele-containing genotypes compared with the MnSOD VV and AV genotypes. DISCUSSION: These results confirm the already established association of MnSOD AA genotype with female breast cancer and further indicate a similar frequency distribution and increased risk in the male population.

INTRODUÇÃO: Uma das diversas rotas metabólicas envolvidas no processo de carcinogênese da mama é o polimorfismo Ala-9Val do gene da superóxido dismutase dependente de manganês, cuja associação com o aumento do risco de câncer de mama em mulheres já é bem estabelecida na literatura. Contudo, não existem estudos envolvendo esse polimorfismo no carcinoma de mama em homens, principalmente devido à baixa prevalência dessa neoplasia. O objetivo deste estudo é analisar o polimorfismo da MnSOD em uma população de homens e mulheres com câncer de mama no sul do Brasil, comparando tais achados com controles saudáveis. MÉTODOS: Foi realizado um estudo de caso-controle em cem pacientes com câncer de mama (11 homens e 89 mulheres) e 370 controles saudáveis. O DNA foi extraído do tecido tumoral emblocado em parafina. O polimorfismo da MnSOD foi determinado por técnicas de PCR-RFLP usando a enzima de restrição Hae III. O teste do qui quadrado foi usado para comparar a distribuição das freqüências dos polimorfismos. RESULTADOS: As freqüências genotípicas dos pacientes com câncer de mama foram AA = 15 por cento; AV = 56 por cento; VV = 29 por cento e dos controles AA = 6,5 por cento; AV = 68,1 por cento e VV = 25,4 por cento. Os pacientes com câncer de mama, tanto as mulheres como os homens, apresentaram freqüências significativamente mais elevadas do genótipo AA quando comparadas aos controles (p = 0,035), sugerindo associação forte desse genótipo com o câncer de mama. O intervalo de confiança foi de 1,393-4,541 (95 por cento) e o risco encontrado foi de 2,15 para indivíduos portadores do genótipo AA, quando comparados com os controles que tinham os genótipos VV e AV da MnSOD. DISCUSSÃO: Esses resultados confirmam a associação já estabelecida do genótipo da MnSOD AA com câncer de mama em mulheres e indicam distribuição de freqüência similar e risco aumentado na população masculina.

Study of the relationship between aluminum overload-induced neurodegeneration and systems of oxidative stress and anti-oxidative stress in mice / 重庆医科大学学报

Objective: To study the roles of the oxidative stress system and the anti-oxidative stress system in neurodegenerative processes induced by aluminum overload in mice.Methods: The neurodegenerative model of mice was established via intragastric administration of AlCl3 solution(element aluminum 400 mg/kg)once a day,with 6 days of week for 3 months.The light-dark test and morris-water-maze were used to evaluate the learning and memory functions.The activities of SOD and SOD2 as well as the contents of MDA in both cortices and hippocampi were measured.Pathomorphological changes of hippocampi were observed.Results: Aluminum overload significantly decreased capacity of passive avoidance learning and memory and spatial recognition,caused the pyknosis and neuron loss of hippocampi,and parallelly increased SOD and SOD2 activities as well as MDA content in cortex and hippocampi,with a tendency of raising first and then gradually decreasing.Conclusions: Aluminum overload markedly causes neurodegeneration and damages learning and memory function in mice,the mechanism of which may be related to the imbalance between oxidative stress and anti-oxidative stress systems in brain induced by aluminum overload.