Aerobic exercise combined with huwentoxin-I upregulates phase-Ⅱ detoxification enzymes to alleviate obstructive jaundice-induced central nervous system injury in mice / 南方医科大学学报

OBJECTIVE@#To explore the effects of aerobic exercise combined with huwentoxin-I (HWTX-I)-mediated Keap1-Nrf2-ARE pathway on phase II detoxification enzymes HO-1 and NQO1 and their protective effects against obstructive jaundice (OJ)-induced central nervous system injury in mice.@*METHODS@#50 male KM mice were randomly divided into blank group (GO), model group (M), aerobic exercise group (T), HWTX-I group (H), and aerobic exercise combined with HWTX-I group (TH). Mouse models of OJ were established with surgical suture for 72 h in the mice in all the groups except for the blank control group. The mice received interventions by aerobic exercise and tail vein injection of HWTX-I (0.05 μg/g) and were assessed by behavioral observation, Clark's neurological function scores, enzyme-linked immunosorbent assay (ELISA), brain tissue Nissl staining, hippocampal tissue Western blotting, and liver tissue mRNA expression profiling and sequencing.@*RESULTS@#The mice in group M had obvious jaundice symptoms after the operation with significantly increased Clark's neurological score ( < 0.01). Compared with those in group M, the mice in group T, group H, and group TH showed significantly decreased serum levels of ALT, AST, TBIL, and TBA ( < 0.01) with increased contents of 5-HT and BDNF and decreased contents of S100B and NSE in the hippocampus ( < 0.01). Synergistic effects between aerobic exercise and HWTX-I were noted on the above parameters except for the liver function indicators. Interventions with aerobic exercise and HWTX-I, alone or in combination, obviously lessened pathologies in the brain tissue induced by OJ, and the combined treatment produced the strongest effect. The treatment also increased the expression levels of Nrf2, HO-1, and NQO1 mRNA and protein in brain tissues ( < 0.01 or 0.05) with a synergistic effect between aerobic exercise and HWTX-I. Illumina high-throughput sequencing showed that the differentially expressed factors participated mainly in such neural regulatory pathways as neuroactive ligand-receptor interaction, GABAergic synapses, dopaminergic synapses, synaptic vesicle circulation, and axon guidance, involving tissue cell neuronal signal transduction, apoptosis inhibition, immune response, and toxicity. Aerobic exercise and HWTX-I synergistically increased the accumulation of the signal pathways related with neuron damage repair and proliferation.@*CONCLUSIONS@#Aerobic exercise combined with HWTX-I can up-regulate the expression of phase Ⅱ detoxification enzymes HO-1 and NQO1 through the Keap1-Nrf2-ARE pathway to protect the central nervous system against OJ-induced damage in mice.

Effects of glutathione s-transferase (GST) M1 and T1 polymorphisms on antioxidant vitamins and oxidative stress-related parameters in Korean subclinical hypertensive subjects after kale juice (Brassica oleracea acephala) supplementation

BACKGROUND/OBJECTIVES: Glutathione s-transferase (GST) is involved in the formation of a multigene family comprising phase II detoxification enzymes, involved in the detoxification of reactive oxygen species. This study evaluated whether daily supplementation with kale juice could modulate levels of plasma antioxidant vitamins and oxidative stress-related parameters. We further examined whether this modulation was affected by combined GSTM1 and T1 polymorphisms. SUBJECTS/METHODS: Totally, 84 subclinical hypertensive patients having systolic blood pressure (BP) over 130 mmHg or diastolic BP over 85 mmHg, received 300 mL of kale juice daily for 6 weeks. Blood samples were drawn before start of study and after completion of 6 weeks. RESULTS: After supplementation, we observed significant decrease in DNA damage and increase in erythrocyte catalase activity in all genotypes. Plasma level of vitamin C was significantly increased in the wild/null and double null genotypes. The plasma levels of β-carotene, erythrocyte glutathione peroxidase activity, and nitric oxide were increased only in the wild/null genotype after kale juice supplementation. CONCLUSIONS: The effect of kale juice was significantly greater in the GSTM1 null genotype and wild/null genotype groups, suggesting possibility of personalized nutritional prescriptions based on personal genetics.

Lymphocyte DNA damage and plasma antioxidant status in Korean subclinical hypertensive patients by glutathione S-transferase polymorphism

BACKGROUND/OBJECTIVES: Glutathione S-transferase (GST) forms a multigene family of phase II detoxification enzymes which are involved in the detoxification of xenobiotics by conjugating substances with glutathione. The aim of this study is to assess the antioxidative status and the degree of DNA damage in the subclinical hypertensive patients in Korea using glutathione S-transferase polymorphisms. SUBJECTS/METHODS: We examined whether DNA damage and antioxidative status show a difference between GSTM1 or GSTT1 genotype in 227 newly diagnosed, untreated (systolic blood pressure (BP) ≥ 130 mmHg or diastolic BP ≥ 85 mmHg) subclinical hypertensive patients and 130 normotensive subjects (systolic BP < 120 mmHg and diastolic BP < 80 mmHg). From the blood of the subjects, the degree of the DNA damage in lymphocyte, the activities of erythrocyte superoxide dismutase, the catalase, and the glutathione peroxidase, the level of glutathione, plasma total radical-trapping antioxidant potential (TRAP), anti-oxidative vitamins, as well as plasma lipid profiles and conjugated diene (CD) were analyzed. RESULTS: Of the 227 subjects studied, 68.3% were GSTM1 null genotype and 66.5% were GSTT1 null genotype. GSTM1 null genotype had an increased risk of hypertension (OR: 2.104, CI: 1.38-3.35), but no significant association in GSTT1 null genotype (OR 0.982, CI: 0.62-1.55). No difference in erythrocyte activities of superoxide dismutase, catalase, or glutathione peroxidase, and plasma TRAP, CD, lipid profiles, and GSH levels were observed between GSTM1 or GSTT1 genotype. Plasma levels of α-tocopherol increased significantly in GSTT1 wild genotype (P < 0.05); however, plasma level of β-carotene increased significantly in GSTT1 null genotype (P < 0.01). DNA damage assessed by the Comet assay was significantly higher in GSTM1 null genotype than wild genotype (P < 0.05). CONCLUSIONS: These results confirm the association between GSTM1 null genotype and risk of hypertension as they suggest that GSTM1 null genotype leads to an increased oxidative stress compared with wild genotype.

Bucillamine prevents cisplatin-induced ototoxicity through induction of glutathione and antioxidant genes

Bucillamine is used for the treatment of rheumatoid arthritis. This study investigated the protective effects of bucillamine against cisplatin-induced damage in auditory cells, the organ of Corti from postnatal rats (P2) and adult Balb/C mice. Cisplatin increases the catalytic activity of caspase-3 and caspase-8 proteases and the production of free radicals, which were significantly suppressed by pretreatment with bucillamine. Bucillamine induces the intranuclear translocation of Nrf2 and thereby increases the expression of gamma-glutamylcysteine synthetase (gamma-GCS) and glutathione synthetase (GSS), which further induces intracellular antioxidant glutathione (GSH), heme oxygenase 1 (HO-1) and superoxide dismutase 2 (SOD2). However, knockdown studies of HO-1 and SOD2 suggest that the protective effect of bucillamine against cisplatin is independent of the enzymatic activity of HO-1 and SOD. Furthermore, pretreatment with bucillamine protects sensory hair cells on organ of Corti explants from cisplatin-induced cytotoxicity concomitantly with inhibition of caspase-3 activation. The auditory-brainstem-evoked response of cisplatin-injected mice shows marked increases in hearing threshold shifts, which was markedly suppressed by pretreatment with bucillamine in vivo. Taken together, bucillamine protects sensory hair cells from cisplatin through a scavenging effect on itself, as well as the induction of intracellular GSH.

The effect of glutathione S-transferase M1 and T1 polymorphisms on blood pressure, blood glucose, and lipid profiles following the supplementation of kale (Brassica oleracea acephala) juice in South Korean subclinical hypertensive patients

BACKGROUND/OBJECTIVES: Glutathione S-transferase (GST) forms a multigene family of phase II detoxification enzymes which are involved in the detoxification of reactive oxygen species. This study examines whether daily supplementation of kale juice can modulate blood pressure (BP), levels of lipid profiles, and blood glucose, and whether this modulation could be affected by the GSTM1 and GSTT1 polymorphisms. SUBJECTS/METHODS: 84 subclinical hypertensive patients showing systolic BP over 130 mmHg or diastolic BP over 85 mmHg received 300 ml/day of kale juice for 6 weeks, and blood samples were collected on 0-week and 6-week in order to evaluate plasma lipid profiles (total cholesterol, triglyceride, HDL-cholesterol, and LDL-cholesterol) and blood glucose. RESULTS: Systolic and diastolic blood pressure was significantly decreased in all patients regardless of their GSTM1 or GSTT1 polymorphisms after kale juice supplementation. Blood glucose level was decreased only in the GSTM1-present genotype, and plasma lipid profiles showed no difference in both the GSTM1-null and GSTM1-present genotypes. In the case of GSTT1, on the other hand, plasma HDL-C was increased and LDL-C was decreased only in the GSTT1-present type, while blood glucose was decreased only in the GSTT1-null genotype. CONCLUSIONS: These findings suggest that the supplementation of kale juice affected blood pressure, lipid profiles, and blood glucose in subclinical hypertensive patients depending on their GST genetic polymorphisms, and the improvement of lipid profiles was mainly greater in the GSTT1-present genotype and the decrease of blood glucose was greater in the GSTM1-present or GSTT1-null genotypes.

Ensaio clínico de fase II da combinação de erlotinibe, quimioterapia e radioterapia em pacientes com carcinoma epidermoide do colo de útero estadiamentos IIB a IIIB / [Phase II clinical trial of the combination of erlotinib, chemotherapy and radiation therapy in patients with cervical squamous cell carcinoma staging IIB a IIIB]

Introdução: o carcinoma epidermoide do colo de útero representa um problema de saúde pública em países em desenvolvimento. Apesar de frequentemente passível de cura quando detectado precocemente, elevado número de pacientes se apresenta com doença localmente avançada ao diagnóstico, cenário clínico associado a resultados terapêuticos insatisfatórios. O padrão atual de tratamento do câncer de colo de útero localmente avançado consiste na combinação de quimioterapia e radioterapia (QT/RXT). Entretanto, após os benefícios obtidos com a adição de quimioterapia baseada em platina à radioterapia, as taxas de cura atingiram um platô. Pacientes com estágios III e IVA apresentaram taxas de sobrevida em cinco anos limitadas a 40 e 15%, respectivamente, enfatizando a necessidade de desenvolvimento de novas estratégias terapêuticas. Estudos préclínicos e clínicos evidenciaram que inibidores do receptor de fator de crescimento epitelial (EGFR) desempenham atividade antitumoral, além de efeito químio e radiossensibilizantes. Erlotinibe (E) é um desses inibidores, com atividade antitumoral pela inibição reversível do EGFR em sua porção tirosino-cinase. Os resultados de estudo de fase I de E combinado a QT/RXT confirmaram a segurança de E na dose de 150 mg/dia. Objetivos: avaliar a segurança e eficácia da combinação de cisplatina, RXT e E em pacientes com carcinoma epidermoide de colo de útero localmente avançado e pesquisa de potenciais marcadores moleculares correlatos (expressão imuno-histoquímica de EGFR e número de cópias do EGFR por FISH). Metodologia: estudo de fase II, unicêntrico, não randomizado. Elegibilidade ­ pacientes com diagnóstico de carcinoma de células escamosas de colo de útero IIB e III; primeira linha terapêutica; ECOG PS 0-2. Tratamento: E 150 mg/dia iniciados uma semana pré-radioterapia e mantidos até o término da braquiterapia; cisplatina ­ 40 mg/m2 /semana/5 semanas; teleterapia (dose de 4.500 cGy) e braquiterapia (quatro inserções semanais de 600 cGy). Biópsia tumoral pré e pós-tratamento foram obtidas para análises moleculares. Avaliação de resposta após três meses do término do tratamento com ressonância magnética, tomografia computadorizada, 18 fluorodeoxiglicose ( 18FFDG), tomografia com emissão de pósitrons (PET-TC) e biópsia. Resultados: foram incluídas 41 pacientes com idade mediana de 44 anos (27-68), estágio IIB (57,9%), IIIA (2,6%) e IIIB (39,5%); 84,3% apresentaram expressão de EGFR, sendo superexpressão em 13%; apenas uma teve amplificação de EGFR por FISH; três foram excluídas; e 38 foram avaliadas para toxicidade; duas não completaram o esquema de tratamento (uma teve a cisplatina suspensa após exibir fenômeno de Raynaud e a segunda apresentou hepatotoxicidade grau IV). A duração mediana do tratamento foi de 77 dias (64-129) e o seguimento mediano atual de 59,3 meses. A combinação terapêutica foi bem tolerada. Todas as 36 pacientes avaliáveis relataram resposta objetiva; 34 (94,4%) exibiram resposta completa; e duas, resposta parcial. As sobrevidas global e livre de progressão foram de 91,7 e 80,6%, e 80 e 73,8% nas medianas de seguimento de 24 e 36 meses, respectivamente. Conclusão: a combinação de RXT/QT/E foi segura e apresentou eficácia promissora em câncer de colo de útero localmente avançado. Este é o primeiro estudo a sugerir o benefício de uma droga-alvo específica neste contexto clínico.

Introduction: Squamous cell cervical carcinoma (CC) represents an important public health concern in developing countries. Although CC is often curable if detected early, a significant number of patients present with locally advanced disease at diagnosis, a clinical scenario associated with suboptimal therapeutic benefits. Combined treatment involving radiotherapy and weekly cisplatin may be considered a reasonable standard of care. After the benefits obtained with the addition of platinum-based chemotherapy to radiotherapy, cure rates of locally advanced cell cervical carcinoma (LACC) have reached a plateau. Therapy results are sub-optimal and patients with stage III and IVA tumors have 5 year survival rates of 40% and 15%, respectively. There is a clear need to explore new strategies to improve prognosis in this group of patients.Pre-clinical and clinical data indicate that EGFR inhibition has antitumor effect itself and potentiates CRT. Erlotinib is a drug that reversibly binds to the the EGFR tyrosine Kinase domain, thereby blocking the signal transduction events, and tumorigenic effects associated with EGFR activation. We previously reported the results from a phase I trial of E administered concurrently with standard chemoradiotherapy for locally advanced cervical cancer that confirmed the safety of E at 150mg/day. Purpose: This phase II trial aims to evaluate the safety and efficacy of erlotinib combined to chemoradiotherapy in locally advanced cervical cancer patients and correlative molecular studies (EGFR IHC and copy number analysis).Methods: Phase II, non-randomised, single institutional.Eligibility - histologically proven stage IIB to IIIB squamous cell cervical carcinoma; no prior therapy; ECOG PS 0-2; adequate end-organ function. Patients received E 150mg/day one week before and combined with C (40mg/m2 , weekly, 5 cycles) and RT (external beam - 4500cGy in 25 fractions, followed by 4 fractions/600cGy/weekly of brachytherapy). Pre and pos treatment tumor tissue was obtained for molecular analysis. Response was assessed after a 3 month-interval with MRI, CT, PET and biopsy.Results: Patient characteristics: median age 44 (27-68), stage IIB 22 (57.9%), IIIA (2.6%) and IIIB 15 (39.5%). 84.3% presented EGFR expression, being 2 or 3 (+) in 13% of them; only one pt presented EGFR amplification by FISH. 41 pts were enrolled and 3 excluded. 38 patients were available for toxicity. Two patients did not complete planned treatment (one presented Raynaud's Syndrome and had C interrupted and one presented grade 4 hepatotoxicity). Of 36 pts who have completed E+CRT, median duration of treatment was 77 (64-129) days and median followup(FUP) is 59.3months(m). Overall E+CRT was well tolerated. Most common grade 3 toxicity was skin rash (13%). Of 36 evaluable for response, 34 patients, (94.4%-IC 95% 79.99- 99.03) had complete response and two patients had partial response. Cumulative OS and PFS survivals were 91.7% and 80.6% and 80% and 73.8%, at the median FUP of 24 and 36 m respectively, superior to historical control. Conclusions: E+CRT is safe and has significant activity in LACC, superior to historical control. To the best of our knowledge, this is the first study to show that a target agent has promising activity in LACC.

Indian studies on genetic polymorphisms and cancer risk.

Genetic influences on cancer development have been extensively investigated during the last decade following publication of human genome sequence. The present review summarizes case-control studies on genetic polymorphisms and cancer risk in Indians. It is observed that the most commonly studied genes in the Indian population included members of phase I and phase II metabolic enzymes. Other than these genes, genetic polymorphisms for cell cycle and apoptosis-related factors, DNA repair enzymes, immune response elements, growth factors, folate metabolizing enzymes, vitamin/hormone receptors, etc., were investigated. Several studies also evidenced a stronger risk for combined genotypes rather than a single polymorphism. Gene-environment interaction was also found to be a determining factor for cancer development in some experiments. Data for single polymorphism and single cancer type, however, was insufficient to validate an association. It appears that much more experiments involving larger sample size, cross-tabulating genetic polymorphisms and environmental factors are required in order to identify genetic markers for different cancers in Indian populations.

Regulation mechanism of triterpenoid components from Prunella asiatica on phase II detoxifying enzymes in vitro and in vivo / 中国中药杂志

To study the effects of triterpenoid components from Prunella asiatica on phase II detoxifying enzymes and protein expression in vitro and in vivo. Normal human bronchial epithelial (NHBE) cell model was used in vitro, and the mouse model of Kunming (KM) mice was used in vivo. CDNB assay was used to measure the activity of GST. NADPH and DCIP was used to detect the activity of NQO1. DTNB colorimetric assay was used to detect GSH. Western blot was use to detect the protein expression of NQO1. We found that triterpenoid components from P. asiatica could increase the activity of GST, NQO1 and GSH in NHBE cells and KM mice. NQO1 protein expression can also be increased in vitro. The study suggests that triterpenoid components from P. asiatica can prevent the lung cancer by regulating the body phase II detoxification enzyme activity and protein expression.

GST (GSTM1, GSTT1, and GSTP1) polymorphisms in the genetic susceptibility of Turkish patients to cervical cancer / 부인종양

OBJECTIVE: This work investigates the role of glutathione S-transferase M1 (GSTM1), glutathione S-transferase T1 (GSTT1), and glutathione S-transferase P1 (GSTP1) enzymes and polymorphisms, which are found in phase II detoxification reactions in the development of cervical cancer. METHODS: This study was conducted with 46 patients diagnosed with cervical cancer and 52 people with no cancer history. Multiplex PCR methods were used to evaluate the GSTM1 and GSTT1 gene polymorphism. However, the GSTP1 (Ile105Val) gene polymorphism was studied using a PCR-RFLP method. The patient and control groups were compared using a chi-square test with p0.05). CONCLUSION: Our results demonstrate that GSTT1, GSTM1, and GSTP1 polymorphisms are not associated with cervical cancer in Turkish patients.

Study on glutathione S-tranferase [GST] inhibition assay by triclabendazole.1: Protoscoleces [Hydatid cyst; Echinococcus granulosus] and sheep liver tissue

Hydatid disease is a term used to refer infection with the methacestode of Echinococcus granulosus parasite in humans, and echinococcusis is restricted to infection with the adult stage in carnivores.Glutathione S-Transferase [GST] represents the major class of detoxification enzymes from helminth parasites such as Echinococcus protoscoleces [PSC] and it is candidate for chemotherapeutic and vaccine design. Therefore, GST of protoscoleces could be a target for evaluation of drug effect as triclabendazole in hydatid cyst. For this purpose, GST enzymes were purified from protoscoleces of hydatid cyst and sheep liver tissue by glutathione affinity chromatography using a wash-batch method and subsequently detected their SDS-PAGE pattern. Afterward, GST specific activity levels were assayed in the whole extract and purified solutions spectrophotometrically at 30 C with reduced glutathione [GSH] and 1-chloro-2, 4-dinitrobenzen [CDNB] substrate. Finally, GST inhibition assay was investigated in the solutions by powder and bolus of triclabendazole. GST fraction as a 26 kDa [MW] band was obtained on SDS-PAGE. The level of GST specific activity in purified solutions was detected 10.24 mmol/min/mg proteins for protoscoleces and 37.84 mmol/min/mg protein for liver tissue. Comparison of the effect of powder and bolus of triclabendazole in solutions revealed inhibition concentration [IC50] 8.71 and 11.16 mg/ml for protoscoleces GST and 8.65 and 9.70 mg/ml for liver tissue GSTs, respectively. These findings suggest the possibility of selective inhibition of protoscoleces. GSTs by triclabendazol in vitro and use of these results for understanding of its molecular effect in vivo