Análise da imunoexpressão de BAX e MMP-9 no líquen plano oral / Analysis of BAX and MMP-9 immunoexpression in oral lichen planus

Introdução: O líquen plano oral é uma doença crônica imunologicamente mediada relativamente comum, que acomete a mucosa oral. Clinicamente, o LPO é classificado em seis padrões bem identificados: placa, reticular, bolhoso, atrófico, papular e erosivo.Sendo os mais comuns oos tipos reticulares e erosivos. A ativação dos linfócitos TCD4+ no LPO, pode induzir os ceratinócitos ao processo de apoptose através da respostaimunológica citotóxica. A proteína Bax desempenha uma função relevante para o processo apoptótico. Deste modo, a presente pesquisa consistiu em um estudo transversal retrospectivo, descritivo, quantitativo e comparativo. Objetivo: Avaliar a expressão imuno-histoquímica das proteínas MMP9 e Bax no LPO. Método: Foram utilizados 43 casos de LPO para análise da imunoexpressão de Bax e MMP-9. Os resultados foram analisados através dos testes estatísticos apropriados e serão considerados significativos, valores onde p<0,05. Resultado: A imunoexpressão de MMP9 foi significativamente maior nos ceratinócitos e quando analisados os subtipos de líquen plano oral, não foram observados diferenças estatísticas entre os tipos reticulares e erosivos para as proteínas analisadas. Conclusões: Com essas observações, infere-se que a alteração na expressão das proteínas estudadas sugere um distúrbio nos mecanismos apoptóticos, os quais estão associados às lesões de LPO, e podemos concluir também que as imunoexpressões dessas proteínas não apresentaram diferença, quando relacionada ao tipo clínico reticular ou erosivo. Com esse resultado pode-se contribuir para um maior entendimento sobre os possíveis mecanismos celulares envolvidos na etiopatogenia dessa lesão (AU).

Background: Oral lichen planus is a relatively common immune-mediated chronic disease that affects the oral mucosa. Clinically, OLP is classified into six well-identified patterns: plaque, reticular, bullous, atrophic, papular, and erosive. The most common being the reticular and erosive types. The activation of TCD4+ lymphocytes in the OLP can induce keratinocytes to the process of apoptosis through the cytotoxic immune response. Thus, the present research consisted of a retrospective, descriptive, quantitative and comparative crosssectional study. Objective: to evaluate the immunohistochemical expression of MMP-9 and Bax proteins in OLP. Methods: We used 20 cases of Inflammatory Fibrous Hyperplasia as control. The results were analyzed through the appropriate statistical tests and will be considered significant, values where p<0.05. Results: The immunoexpression of MMP-9 was significantly higher in keratinocytes and when the subtypes of oral lichen planus were analyzed, no statistical differences were observed between the reticular and erosive types for the proteins analyzed. Conclusions: With these observations, it is inferred that the alteration in the expression of the studied proteins suggests a disturbance in the proliferative and apoptotic mechanisms, which are associated with a pathological behavior of the oral mucosa, and consequently with a repercussion on the lesions of OLP, and we can also conclude that the immunoexpression of these proteins had no difference, when related to the reticular or erosive clinical type. This research aims to contribute to a greater understanding of the possible cellular mechanisms involved in the etiopathogenesis of this lesion, thus enabling the understanding of the clinical aspects of the pathology (AU).

Effect and mechanism of allicin combined with 5-fluorouracil on proliferation and apoptosis of the MEC-1 cell line in mucoepidermoid carcinoma / 口腔疾病防治

Objective @#To investigate the effect and mechanism of allicin combined with 5-FU on proliferation inhibition and apoptosis of the mucoepidermoid carcinoma MEC-1 cell line in mucoepidermoid carcinoma in order to provide the corresponding basis for subsequent clinical drug application.@*Methods @# MEC-1 cells in the logarithmic growth phase were randomly divided into control groups and experimental groups. The control groups were PBS groups containing 0.1% DMSO, while the experimental groups were the allicin group, 5-FU group and combined drug group (the allicin combined with the 5-FU group). The proliferation inhibition rates of allicin, 5-FU and allicin combined with 5-FU in MEC-1 cells were detected by the CCK8 method at different concentrations (0, 25, 50, and 75 mg/L) for 24 h, and the IC50 value of allicin and 5-FU after 24 hours was calculated. The apoptotic rate of MEC-1 cells treated with allicin, 5-FU and allicin combined with 5-FU at different concentrations (0, 25, 50, and 75 mg/L) for 24 hours was measured by flow cytometry. The expression of Bax and Bcl-2 protein was determined by Western blot analysis of the IC50 concentration of allicin and 5-FU alone and in combination with MEC-1 cells for 24 hours. @*Results@#The growth inhibition rate and apoptosis rate of MEC-1 cells in the combined drug group were higher than those in the allicin group and the 5-FU alone group (P < 0.01). Allicin and 5-FU alone and in combination downregulated Bcl-2 protein and upregulated Bax protein expression, and the combined drug group had the largest ratio of Bax/Bcl-2 (P < 0.05). @*Conclusion @#Allicin and 5-FU both alone and in combination can inhibit the proliferation of and induce apoptosis in MEC-1 cells, and allicin can enhance the apoptosis of 5-FU in MEC-1 cells, which may be related to the apoptosis of the mitochondrial pathway.

Auraptene has neuroprotective and memory enhancing effects in a rat model of Alzheimer’s disease

@#Background: Auraptene is a simple coumarin that exhibits multiple protective activities in the brain. Alzheimer’s disease is a complex, multifactorial, and progressive neurodegenerative disease. Microinjection of the β-amyloid peptide (Aβ) into the hippocampus of rat has been recognized as a reliable and stable animal model of Alzheimer’s disease, which mimics the memory deficits. In the present study, the memory enhancing effects of auraptene were studied in rats that Aβ was injected into their hippocampus to create a model of Alzheimer’s disease. Methods: Different doses of auraptene (5, 10 and 25 mg/kg) were administered intraperitoneally to male Wistar rats. The spatial memory performance was tested by Morris water maze after Alzheimer`s induction. The hippocampal expression of pro-apoptotic Bax and anti-apoptotic Bcl-2 proteins were calculated for evaluating the neuroprotective and anti-apoptotic effects of Auraptene in the brain tissue. Results: In comparison with the control group, auraptene significantly decreased the escape latency time in the treated rats. In addition, auraptene increased the percentage of time spent and traveled pathway in the target quadrant. Molecular data showed that auraptene attenuated the Bax/Bcl-2 ratio in the hippocampus of rats. Conclusion: This study demonstrated the memory enhancing effect of Aur after Aβ injection, which could be through inhibiting the apoptotic pathways in the hippocampus of rats.

Study on the mechanism of Sorafenib combined with Bevacizumab on the tumor inhibition of cervical cancer cell line U14 transplantation in mice / 国际外科学杂志

Objective According to effect of Sorafenib combined Bevacizumab in mice U14 cervical cancer cell lines,observe the changes of cell lines of transplanted tumor U14 cell structure and adjustment effect of Bcl-2 and Bax protein expression.Methods Sixty female 615 mice of healthy inbreeding line,aged from 4 to 6 weeks,average weight is 20.4 g,range from 18 to 25 g.The animals were divided into blank control group (group A)with 10 mice,and U14 cervical cancer cell lines by vaccinating mice a tumor-burdened mice model was established,on the 6th day after subcutaneous inoculation of U14,40 tumor-burdened mice with tumor diameter ≥ 5 mm were selected.The mice were divided into 5 groups according to the completely randomized grouping method:model group (group B),Sorafenib group (group C),low-dose Bevacizumab combined with Sorafenib group (group D),the middle dose Bevacizumab combined with Sorafenib group (group E),and the high dose Bevacizumab combined with Sorafenib group (group F),8 mice in each group.The treatment regimen consisted of 1 ml of 0.9％ sodium chloride solution in group A and group B,Sorafenib 12 mg/kg in group C,and Bevacizumab 2.5 mg/kg + Sorafenib 12 mg/kg in group D,Bevacizumab 5 mg/kg + Sorafenib 12 mg/kg in group E,and Bevacizumab 10 mg/ kg + Sorafenib 12 mg/kg in group F.All mice were injected intraperitoneally and sacrificed by dislocation 24 days later.The tumor weight (g) was measured every 6 days,and the body weight of each group was observed,and calculated the tumor inhibition rate of each group of mice.The histopathological changes of the transplanted mice were observed by hematoxylin-eosin staining.The transplantation of each group of mice was observed by electron microscope.Morphological changes of tumor tissue;the expression of Bcl-2 and Bax protein in each group were observed by immunohistochemistry.The measurement data were expressed as mean standard deviation (Mean ± SD),univariate analysis of variance was used for inter-group comparison,and LSD method was used for pairwise comparison.Results At the beginning of the intraperitoneal injection of the drug,the body weight of the mice began to increase slowly and continuously,and the trend of the mice in each group was basically the same.The tumor inhibition rate of group C was 8.02％,4.92％ in group D,11.10％ in group E,and 5.42％ in group F.Group E had the highest tumor inhibition rate and the best effect.The difference in tumor weight between group A and the other groups was statistically significant (P ＜ 0.05).The results of electron microscopy showed that the cell structure changes in C,D,E and F groups were similar,and the apoptosis of tumor cells appeared after treatment.The apoptosis of group E was the best.The apoptosis-related proteins in group C,D,E and F were observed.The expression of Bax was up-regulated,and the positive number in group E was the highest.The expression of proto-oncogene Bcl-2 was down-regulated in group C,D,E and F,and the number of positive in group E was the least,and the difference was statistically significant (P ＜ 0.05).Conclusions Sorafenib combined with Bevacizumab can promote tumor cell apoptosis by up-regulating Bax and down-regulating Bcl-2 protein expression.Among them,Sorafenib combined with Bevacizumab at medium dose promotes tumor cell apoptosis better than other methods,providing a basis for clinical treatment.

Bax protein structure and related research progress of eutectic structure / 国际生物医学工程杂志

Apoptosis involves multiple signaling pathways. The intrinsic signaling pathway is the mitochondrial apoptotic pathway, which is caused by a series of processes. First, the pro-apoptotic factors such as Bax are activated by signaling molecules and transfer to the mitochondrial outer membrane forming protein pores, thus the mitochondrial membrane permeability is affected, and then the downstream caspase-9 is activated and the apoptosis initiation is induced by releasing cytochrome C. In order to explore the apoptosis initiation activated by small molecules, the specific structural changes of Bax in apoptosis were studied. The results showed that there is a hydrophobic pocket structure near the C-terminal S184 of the Bax protein. This structure can be combined with certain small molecular substances specifically remove phosphorylation S184, and regulate Bax protein to promote apoptosis activity. At present, the nuclear magnetic structure of Bax protein has been obtained and the crystal structure has not been revealed. The eutectic structure formed by corresponding with other proteins in the Bcl-2 family has been resolved, which can be used to study the interactions between proteins and to understand the specific structural changes in the formation of heterologous dimers during apoptosis, site changes, etc. In this paper, the Bax protein structure resolved by nuclear magnetic structure was reviewed to learn the change of the sites in the induced apoptosis so as to promote the research on apoptosis initiation.

Effect and mechanism of biochanin A on rat hippocampal neurons injuries impaired by H2O2 / 中国药学杂志

OBJECTIVE: To observe the effect of Biochanin A (Bio A) on rat hippocampal neurons injuries impaired by H2O2 and Bcl-2, Bax and caspase-3 protein expression. METHODS: Hippocampal neurons of newly born rat were cultured in vivo and were injured by H2O2. Different concentration of Bio A on cell viability was measured by CCK-8; cell apoptosis rate was tested by flow cytometry analysis; Bcl-2, Bax and caspase-3 protein expression were tested by Western blot method. RESULTS: Compared with the model group, Bio A significantly improved hippocampal neurons cell viability and inhibited cell apoptosis and necrosis. In addition, Bio A clearly enhanced the protein expression of Bcl-2 and decrease the protein expression of Bax and caspase-3. CONCLUSION: The protective effect of Bio A on rat hippocampal neurons injuries impaired by H2O2 may be related to the inhibition of hippocampal neurons apoptosis. The mechanism of Bio A against cell apoptosis may involve the increased Bcl-2 protein expression and reduced Bax, caspase-3 protein expression.

Dipeptidyl peptidase-4 inhibitor impacts the expression of Bcl-2 and Bax throughγamino acid bwtyric acid in pancreas islets / 中华内分泌代谢杂志

Objective To explore the effects of dipeptidyl peptidase ( DPP-4 ) inhibitor on proteins expression of Bcl-2 and Bax of islet β-cells through increasing the expression of islet γ amino acid butyric acid ( GABA) . Methods A total of 50 rats of clean grade were studied. Among them, ten rats were randomly selected as normal controls, the remaining forty rats were fed with high-fat diet and then intraperitoneal injection with streptozotocin, the diabetic rats models were then established. Rats were randomly divided into three groups:i. e. diabetic control group, DPP-4 inhibitor group, and antagonist group ( DPP-4 inhibitor and GABA receptor antagonist). Six weeks later, blood glucose, serum insulin, glucagon, and the proteins expression of GABA, Bcl-2, and Bax of islet β-cells were measured. Results ( 1 ) Compared with diabetic control group, serum insulin was increased(P<0.05),bloodglucoseandserumglucagonweredecreasedinDPP-4inhibitorgroup(P<0.05). (2) Compared with DPP-4 inhibitor group, serum insulin was decreased(P<0. 05), blood glucose and serum glucagon were increased(P<0. 05) in antagonist group. (3) Compared with diabetic control group, the expression of GABA was increased(P<0. 05), the expression of Bcl-2 protein was increased (P<0. 05) in pancreatic β-cells in DPP-4 inhibitor group. ( 4 ) Compared with diabetic control group, the expression of GABA in pancreatic β-cells was increased in antagonist group(P<0. 05) . Compared with DPP-4 inhibitor group, the expression of Bax protein in pancreaticβ-cells was increased in antagonist group(P<0. 05) , while the expression of Bcl-2 protein was decreased (P<0. 05). Conclusions DPP-4 inhibitor could increase the secretion of insulin, decrease the secretion of glucagon, up-regulate expression of anti-apoptosis protein Bcl-2, and down-regulate expression of apoptosis protein Bax in pancreatic β-cells through increasing the expression of GABA, inhibiting pancreatic β-cells apoptosis and protecting the damagedβ-cells in type 2 diabetic rats.

Radiosensitizing effect and mechanism of tanshinone I in mice bearing lewis lung carcinoma / 中国药学杂志

OBJECTIVE: To investigate the mechanism of inhibition and radiosensitization effects of combining treatment with tanshinone I and(irradiation) IR on mice bearing Lewis lung carcinoma(LLC). METHODS: The models of LLC in C57BL/6 mice were established via subcutaneous injection of mouse LLC cells, and divided into model control group, radiation alone group, Tan I (40 mg · kg-1) group, 5-Fu + IR group, Tan I (10, 20, 40 mg · kg-1) + IR groups. After irradiated, the relative volume of tumor was observed, the delay time of tumor growth and enhancement factor (EF) was calculated. After stripped, the inhibition rate was calculated. Cell apoptosis index( AI) was in vestigated by TUNEL staining. The protein expressions of Bcl-2 and Bax were detected by Western blot. RESULTS The different concentrations of Tan I (low, medium and high) + IR groups inhibited the tumor growth significantly, and the inhibition rate was 27.14%, 38.46%, 59.78%, respectively, which were significant difference as compared with IR group (P < 0.01). And also had the significant radiosensitizing effect, the enhancement factor was 1.09, 1.76, 2.03, respectively. The AI was 51.3% in Tan I (H) + IR group, which was significant difference as compared with IR group(P < 0.05). The expression of Bcl-2 protein was decreased, and Bax proteins was increased by combining treatment with Tan I and IR. CONCLUSION Combining treatment with Tan I and IR inhibited the tumor growth and had radiosensitizing effects in mice bearing LLC. One of the mechanism may be that it might induce apoptosis by regulated the expression of Bcl-2/Bax protein, so cells were more sensitive to radiation.

Effect of Vaccinium vitis procyanidin on regulation of glioma cell growth / 中国病理生理杂志

AIM:To explore the effect of Vaccinium vitis procyanidin on the growth of glioma cells .METH-ODS:Glioma C6 cells were cultured and divided into control and 10, 20 and 40μg/L Vaccinium vitis procyanidin groups . The influence of Vaccinium vitis procyanidin on the growth of C 6 cells was measured by MTT assay and the observation un-der inverted microscope .The apoptotic rate was detected by Annexin V/PI staining .The protein expression of Bcl-2 and Bax was determined by immunocytochemistry .The protein levels of Bcl-2, Bax and caspase-3 were also examined by West-ern blotting .RESULTS:The growth of C6 glioma cells was inhibited by Vaccinium vitis procyanidin at concentrations of 10, 20 and 40 μg/L.The growth was significantly inhibited in 40 μg/L Vaccinium vitis procyanidin group at 24 h and 48 h, and in 20 and 40 μg/L Vaccinium vitis procyanidin groups at 72 h (P<0.01).The density of the cells was decreased when the concentration of Vaccinium vitis procyanidin increased .The apoptotic rate was increased when the concentration of Vaccinium vitis procyanidin increased either .The expression of Bcl-2 was decreased and Bax was increased after 10, 20 and 40 μg/L Vaccinium vitis procyanidin treatments .The ratio of Bax/Bcl-2 was increased when the dose of Vaccinium vitis pro-cyanidin increased (P<0.05 or P<0.01).The expression of Bcl-2 was decreased (P<0.01), and Bax and caspase-3 were increased after 10, 20 and 40 μg/L Vaccinium vitis procyanidin treatments .The ratio of Bax/Bcl-2 was increased when the dose of Vaccinium vitis procyanidin increased (P<0.01).CONCLUSION:Vaccinium vitis procyanidin inhibits the growth of glioma cells by down-regulating Bcl-2 protein and up-regulating Bax protein to activate caspase-3, thus indu-cing apoptosis .

Protective effect of nimodipine on neuron of rats with focal cerebral ischemia-reperfusion inj ury and its mechanism / 吉林大学学报(医学版)

Objective To investigate the protective effect of nimodipine on neuron of the rats with focal cerebral ischemia-reperfusion injury and the expressions of Bax and Bcl-2,and to clarify their mechanisms.Methods The focal cerebral-ischemia reperfusion model was induced by the middle cerebral artery occlusion(MCAO)method. 30 male Wistar rats were randomly divided into sham operation,model,and nimodipine groups(n=10).The neurological deficit score was performed after 2 h ischemia following 2 h reperfusion.The infarction was observed by TUNEL staining and the expressions of Bax and Bcl-2 were detected by SP immunohistochemistry method. Results Compared with model group, the number of apoptotic cells of the rats in nimodipine group was significantly decreased(P<0.05),the expression of Bax was significantly decreased (P<0.05),and the Bcl-2 expression was increased significantly(P<0.05).The morphological examination showed that the neurons of the rats in model group had serious necrosis and edema while the number of dead cells in nimodipine treatment group was reduced and the edema was improved.Conclusion Nimodipine has a protective effect on brain tissue of the rats with focal cerebral ischemia-reperfusion inj ury, which is closely related to the down-regulation of Bax and up-regulation of Bcl-2 and inhibition of the apoptosis of neuron.

Effects of matrine on apoptosis and related protein expression in human medulloblastoma D341 cells / 中国病理生理杂志

AIM:To investigate matrine-induced apoptosis of human medulloblastoma D 341 cells and the ex-pression of Bax , Bcl-2, serine/threonine kinase Akt and phosphorylated Akt ( p-Akt) in vitro.METHODS: D341 cells were divided into experimental groups ( added with matrine at different concentrations ) and control group ( under the same conditions without matrine).The proliferation of D341 cells was analyzed by CCK-8 assay.Apoptosis was detected by An-nexin V-FITC/PI double staining and the expression of Bax , Bcl-2, Akt and p-Akt was detected by Western blotting .RE-SULTS:Matrine significantly inhibited the proliferation of D 341 cells and increased the apoptosis in a dose-and time-de-pendent manner .The cell apoptosis was characterized by chromatin condensation with margination of chromatin to the nu -clear membrane , increased when and larger cytoplasmic vacuoles , and formation of apoptotic body after treatment with ma-trine.The expression of Bax increased , while the expression of Bcl-2 and p-Akt decreased when the drug concentration gradually increased .CONCLUSION:Matrine induces the apoptosis of human medulloblastoma D 341 cells in vitro by acti-vation of Bax, down-regulation of Bcl-2 and reduction of p-Akt expression level in the PI3K/Akt signaling pathway.

Influence of low level, long-term arsenite exposure on HaCat cell apoptosis and the expression of Bcl-2 and Bax proteins / 中华地方病学杂志

Objective To investigate the influence of low-dose,long-term arsenite exposure on keratinocyte (HaCat) cell apoptosis and the expression of Bcl-2 and Bax proteins.Methods HaCat cells were exposed to different concentrations of NaAsO2[0(control group),0.05,0.10 μmol/L] for 15 weeks.A total of 10 000 cells in each group were measured to detect the level of apoptosis by flow cytometry.The protein expressions of Bcl-2 and Bax were determined by Western blotting method.Results The cell apoptosis rate was significantly different between groups (F =17.19,P ＜ 0.05).Compared with that of the control [(1.42 ± 0.13)％],the apoptosis rates of the 0.05 and 0.10 μmol/L groups [(1.23 ± 0.08)％ and (1.04 ± 0.06)％] decreased significantly (all P ＜ 0.05); the 0.10 μmol/L group was significantly lower than that of the 0.05 μmol/L group(P ＜ 0.05).The protein expressions of Bcl-2 and Bax were significantly different between groups (F=107.38,346.45,all P＜ 0.05).The protein expression of Bcl-2 of the 0.05 and 0.10 μmol/L groups were (143.89 ± 7.74)％ and (199.96 ± 12.18)％,respectively,which were significantly higher than that of the control group[(100.00 ± 1.45)％,all P ＜ 0.05] ; the 0.10 μmol/L group was significantly higher than that of the 0.05 μmol/L group (P ＜ 0.05).The protein expression of Bax of the 0.05 and 0.10 μmol/L groups were (70.78 ± 1.53)％ and (54.80 ± 1.34)％,respectively,which were markedly lower than that of the control group[(100.00 ± 3.09)％,all P ＜ 0.05]; the 0.10 μmoL/L group was significantly lower than that of the 0.05 μmol/L group(P ＜ 0.05).Conclusion Decreased level of apoptosis induced by low level and long-term arsenic exposure may be associated with increased expression of Bcl-2 protein and decreased expression of Bax protein.

Effect of Shenlongtang on Learning and Memory Ability and Expression of Bcl-2 and Bax in Hippocampus of Cerebral Ischaemia Rats / 中国中医药信息杂志

Objective To investigate the effect of Shenlongtang decoction on learning and memory ability and expression of Bcl-2 and Bax on rats hippocampus with cerebral ischemia model. Methods The cerebral ischemia animal model was established by two-vessel occlusion. Fifty rats were randomly divided into control group, model group, pisacetam group, Shenlongtang groups (low and high dosage). After modeling, rats were administrated with corresponding drugs for 28 days, the learning and memory ability was tested with Morris water maze, the number of positive cells and the integral optical density of the immunostaining on Bcl-2 and Bax protein expression in the hippocampus was detected. Results With Morris water maze test, the latency time increased significantly and times of searching submerged platform decreased significantly in the model group, the difference was significant compared with control group (P<0.01). Compared with model group, the latency time of Shenlongtang high dosage group decreased significantly and times of searching submerged platform increased significantly (P<0.05). Compared with model group, the expression of positive cells number of Bax protein expression in the hippocampus area was significantly increased (P<0.05), the Bcl-2 protein expression in the hippocampus region was significantly decreased (P<0.01). Conclusion Shenlongtang could significantly improve the learning and memory ability of cerebral ischemia rats, the mechanism may be related to regulating the expression of Bcl-2 and Bax.

Effect of genistein on the expression of Bax and Bcl-2 protein in PC12 cells transfected with mutant APP695 type / 中华行为医学与脑科学杂志

Objective To investigate the effect of genistein on the expression of Bax and Bcl-2 protein in PC12 cells transfected with mutant APP695 Type.Methods PC12 cells were transfected with pIRES2-EGFP plasmid or pIRES2-EGFP/APP695MT expression plasmid,and then divided into normal control group,Unladen group,APP695 transfected group,GST treatment group(5 μmol/L,15 μmol/L).The Bax and Bcl-2 protein expression in PC12 cells was measured by Immunocytochemistry SABC and Western-blot.Results Immunocyto-chemistry SABC showed that:compared with the normal control group,the expression of bax protein in APP695 transfected group was strongly increased((78.02 ± 1.26) ％,P＜0.01)) and the expression of bcl-2 protein was very weak oppositely ((15.40 ± 0.72) ％,P＜0.01)).Compared with APP695 transfected group,the expression of bax protein in cells of 15umol/L GST treatment group was significantly decreased((22.35 ± 0.49) ％,P＜0.01))and the expression of bcl-2 protein in these cells was strongly increased ((68.11 ± 0.24) ％,P ＜0.01)).Western-Blot showed that:compared with the normal control group,the expression of bax protein in APP695 transfected group was markedly increased and the expression of Bcl-2 protein was significantly decreased.Compared with the APP695 transfected group,the expression of Bax protein was markedly decreased and the expression of Bcl-2 protein was significantly increased in the cells of the GST treatment groups.Conclusion GST can reduce the expression of bax protein and the increased expression of Bcl-2 protein has protective effect on PC12 cells transfected with APP696MT.

Proteínas pro-apoptóticas y anti-apoptóticas como factores de pronóstico en Linfoma B difuso de célula grande en adultos / Pro-apoptotic and anti-apoptotic proteins as prognostic factors in diffuse large B-cell lymphoma in adults / Proteínas pró-apoptóticas e anti-apoptóticas como fatores de prognóstico no Linfoma B difuso de célula grande em adultos

Objetivo. El propósito de este estudio fue evaluar en pacientes con LBDCG la expresión de las proteínas anti-apoptóticas Bcl-2 y Bcl-X L y de las proteínas pro-apoptóticas Bad y Bax y su asociación con la supervivencia. Materiales y métodos. Se analizaron biopsias de 28 pacientes con diagnóstico de LBDCG. La expresión de los reguladores apoptóticos se evaluó mediante western blot. La asociación entre la expresión de las proteínas y la supervivencia fue analizada mediante el método de Kaplan-Meier y la prueba log-rank. Resultados. Las proteínas Bcl-2, Bak, Bad y Bcl-xL se encontraron expresadas en el 78,8%; 71,4%; 64,3% y 50% de los casos de LBDCG respectivamente. No encontramos asociación entre la presencia de las proteínas o sus niveles de expresión y la supervivencia total. La presencia de las proteínas Bad y Bcl-xL se asoció con una mayor supervivencia libre de enfermedad (33,3% vs. 20,0%, p LR test= 0,003; 42,9% vs. 14,3%, p LR test= 0,03 respectivamente). Niveles altos de expresión de Bad y de Bcl-X L se asociaron con una supervivencia libre de enfermedad mayor (35,7% vs. 21,4%, p LR test= 0,012 y 42,9% vs. 14,3%, p LR test= 0,045 respectivamente). Conclusión. Dado que la expresión de la proteína Bad en los tumores se asoció con una mayor supervivencia libre de enfermedad, los pacientes con bajos niveles de expresión de esta proteína podrían ser beneficiados en un futuro con terapias orientadas a inhibir las moléculas anti apoptóticas Bcl-xL y Bcl-2 mediante el empleo de moléculas que se unen específicamente al dominio BH3.

Objective. Our purpose was to evaluate the expression of antiapoptotic proteins Bcl-2 and Bcl-xL, and pro-apoptotic proteins Bad and Bax and their association with survival, in patients with DLBCL. Materials and methods. We analyzed biopsies from 28 patients diagnosed with DLBCL. The expression of the apoptotic regulators was assessed by western blot. The association between protein expression and survival was analyzed by the Kaplan-Meier method and the log-rank test. Results. Bcl-2, Bak, Bad and Bcl-xL proteins were expressed in 78.8, 71.4, 64.3 and 50% of the DLBCL cases, respectively. We found no association between the presence of proteins or their expression levels and overall survival. Both Bad and Bcl-xL were associated with higher disease-free survival (33.3% vs. 20.0%, p LR test= 0,003; 42.9% vs. 14.3%, p LR test= 0.03, respectively). High expression levels of Bad and Bcl-xL were associated with a higher disease-free survival (35.7% vs. 21.4%, p LR test= 0.012 y 42.9% vs. 14.3%, p LR test= 0.045, respectively). Conclusion. Given that expression of the Bad protein in tumors was related to a higher disease-free survival, patients with low expression levels of Bad could be candidates in future therapies oriented towards the inhibition of the anti-apoptotic proteins Bcl-xL and Bcl-2 by using molecules that bind specifically to the BH3 domain.

Objetivo. O objetivo deste estudo foi avaliar em pacientes com LBDCG a expressão das proteínas anti-apoptótica Bcl-2 e Bcl-X L e das proteínas pró-apoptóticas Bad e Bax e sua associação com a sobrevivência. Materiais e métodos. Foram analisadas biópsias de 28 pacientes com diagnóstico de LBDCG. A expressão de reguladores de apoptose foi avaliada por Western blot. A associação entre a expressão das proteínas e a sobrevivência foi analisada pelo método de Kaplan-Meier e o teste log-rank. Resultados. As proteína Bcl-2, Bak, Bad e Bcl-xL foram expressas em 78,8%, 71,4%, 64,3% e 50% dos casos de LBDCG, respectivamente. Nós não encontramos associação entre a presença das proteínas ou de seus níveis de expressão e a sobrevivência total. A presença das proteínas Bad e Bcl-xL foi associada à maior sobrevivência livre da doença (33,3% vs. 20,0%, p LR teste= 0,003; 42,9% vs. 14,3%, p LR teste= 0,03, respectivamente). Altos níveis de expressão de Bad e de Bcl-X L foram associados com uma sobrevivência livre da doença maior (35,7% vs. 21,4%, p LR teste= 0,012 e 42,9% vs. 14,3%, p LR teste= 0,045, respectivamente). Conclusão. Uma vez que a expressão da proteína Bad em tumores foi associada com uma maior sobrevivência livre de doença, os pacientes com baixos níveis de expressão dessa proteína poderiam ser beneficiados num futuro com terapias orientadas a inibir as moléculas anti-apoptóticas Bcl-xL e Bcl-2 utilizando moléculas que se ligam especificamente ao domínio BH3.

Induction of the Intrinsic Apoptotic Pathway by 3-Deazaadenosine Is Mediated by BAX Activation in HL-60 Cells

3-Deazaadenosine (DZA), a potent inhibitor of S-adenosylhomocysteine hydrolase, was previously proposed to induce intrinsic apoptosis in human leukemic cells. In the present study, we analyzed the mechanism underlying the DZA-induced intrinsic apoptotic pathway. DZA activated typical caspase-dependent apoptosis in HL-60 cells, as demonstrated by an accumulation of hypo-diploidic cells, the processing of multiple procaspases and an inhibitory effect of z-VAD-Fmk on this cell death. During DZA-induced apoptosis, cytochrome c (cyt c) was released into the cytosol. This was neither prevented by z-VAD-Fmk and nor was it associated with the dissipation of mitochondrial membrane potential (DeltaPsim). Prior to the release of cyt c, BAX was translocated from the cytosol to mitochondria and underwent oligomerization. Finally, the overexpression of BCL-XL protected HL-60 cells from apoptosis by blocking both the cyt c release and BAX oligomerization. Collectively, these findings suggest that DZA may activate intrinsic apoptosis by stimulating BAX activation and thereby the release of cyt c.

Effect of 18F-2-deoxy-2-fluoro-d-glucose on proliferation of Lewis lung cancer cell / 中华放射医学与防护杂志

Objective To investigate the influence of 18F-FDG on the proliferation of Lewis lung cancer cell line,and to elucidate its possible mechanism.Methods Morphological changes of cells after culture for 24 h at different concentrations of 0,0.37,1.85,3.70 and 7.4 (×106) Bq/ml of 18F-FDG were observed by using inverted microscopy and electron microscopy.The apoptosis and phase distribution of cell cycle of irradiated cells were analyzed with flow cytometry.DNA synthesis of irradiated cells was assayed by 3H-TdR incorporation.Lipid peroxidation was measured by chromometry and expression of Bcl-2 and Bax protein was measured by immunohistochemical technique.Results Exposed to (0-7.40) × 106Bq/ml of 18F-FDG for 24 h,the cumulative absorbed doses delivered to cells in five groups were 0,0.11,0.55,1.10 and 2.20 Gy,respectively.Irradiated cells showed morphological changes of apoptosis.The apoptosis rate of irradiated cells was increased from (4.05 ± 0.01)% to (25.6 ± 0.28) % (t = 188,P＜0.01).3H-TdR incorporation rate was decreased from 100% to(22.0 ± 0.51)% (t =27.6,P ＜0.05).The levels of M DA in cells were augmented from (0.08 ± 0.03) to (0.67 ± 0.12) μmol/L (t =11.7,P ＜ 0.01).Cell cycle arrest was found in G2/M phase with the increasing doses from 0 to 2.20 Gy.The expression of Bcl-2 protein was decreased while that of Bax protein increased.Conclusions 18F-FDG could induce the apoptosis of cells and inhibit the proliferation of cells.

Effects of mild hypothermia performed at different times on levels of glutamate, Bcl-2 and Bax during global cerebral ischemia-repersusion in rats / 中华麻醉学杂志

Objective To evaluate the effects of mild hypothermia performed at different times on the levels of glutamate, Bcl-2 and Bax during global cerebral ischemia-reperfsusion (I/R) in rats. Methods Twenty-four male SD rats weighing 230-270 g were randomly divided into 4 groups according to the time at which mild hypothermia was performed ( n =6 each): group A, B, C and D. Global cerebral ischemia was produced by occlusion of 4 vessels (cauterization of bilateral vertebral arteries and occlusion of bilateral common carotid arteries) in the 4 groups. In group B, C and D, the nasopharyngeal temperature was reduced to 32.5-33.5 ℃ and maintained for 1 h. Ischemia was performed after termination of cooling in group B. While ischemia was performed, cooling was started in group C. While reperfusion was performed, cooling was started in group D. Rewarming was started after termination of cooling in group B, C and D. Samples of dialysate in hippocampal CA1 area were collected every 10 min during 100 min reperfusion for determination of glutamate concentrations by high-performance capillary electrophoresis with laser-induced fluorescence detection ( HPCE-LIF). The brain tissues were taken at 3 h of reperfusion for determination of the expression of Bax and Bcl-2 in hippocampal CA1 area, and the ratio of Bcl-2/Bax was calculated. Results Compared with group A, the glutamate concentrations were significantly decreased, the Bcl-2 expression was up-regulated, the Bax expression was down-regulated and the ratio of Bcl-2/Bax was increased in the other thee groups ( P ＜ 0.05). Compared with group B, the Bcl-2 expression was significantly down-regulated, the Bax expression was up-regulated and the ratio of Bcl-2/Bax was decreased in group C ( P ＜0.05). The glutamate concentrations were significantly increased, the Bax expression was up-regulated and the ratio of Bcl-2/Bax was decreased in group D compared with group C ( P ＜ 0.05 ). Conclusion The earlier cooling is performed, the better the cerebral protective effect is during global cerebral I/R in rats.

Radon induced pulmonary lesion and expression of P53 and Bcl-2,Bax in mice / 中华放射医学与防护杂志

Objective To establish the mice model of lung injury induced by radon,and to observe the changes of pulmonary lesion at different doses and to analyze the influence of radon exposure on P53 and Bcl-2、Bax expression in lung tissue.Methods Mice were exposed to radon of 30 and 60 WLM,respectively.Apoptosis was detected by terrainal deoxynucleotidy transferse-mediated dUTP-biotin nick end labeling(TUNEL).The expressions of P53,Bcl-2 and Bax protein were observed by immunohistochemistry and Western blot.Results Compared with those in the control group,the apoptotic indexes increased significantly in the 30 and 60 WLM groups(t=18.11,-10.30,P＜0.05).The protein expression of P53 was significantly increased(t=-11.08,P＜0.05).The expression levels of P53 and Bax were remarkably inereased(t=-7.00,-2.52,P＜0.05),while the expression of Bcl-2 protein was significantly decreased in the 60 WLM group(t=4.36,P＜0.05).But Bcl-2,Bax expression was decreased in both 30 WLM and 60 WLM groups(t=2.78,4.07,P＜0.05).Conclusions Radon could induce pulmonary lesion of mice.It may be involved in the regulation of apoptosis of pulmonary lesion by the P53,Bcl-2、Bax pathway.

Effects of Ethyl Pyruvate on Myocardial Apoptosis and Expression of Bcl-2 and Bax Proteins after Ischemia-reperfusion in Rats / 华中科技大学学报（医学）（英德文版）

In order to study the effects of ethyl pyruvate on cardiomyocyte apoptosis following ischemia/reperfusion (I/R) in vitro and the expression of Bcl-2 and Bax proteins, isolated rat hearts were perfused in a Langendorff model. Twenty-four rats were randomly divided into 3 groups (n=8 in each group): control group was perfused for 120min. In the I/R group, after 30min stabilization the injury was induced by 30min global ischemia followed by 60min reperfusion. Ethyl pyruvate (EP) group was set up with the same protocol as I/R group except that it was supplied with 2mmol/L EP 15min before ischemia and throughout reperfusion. Myocardial malonaldehyde (MDA) content Was measured. Myocardial apoptotic index (AI) was tested by terminal deoxynucleotidyl transferase mediated dUTP nick end labeling (TUNEL) method. The expression of anti-apoptotic protein Bcl-2 and pro-apoptotic protein Bax in cardiac myocytes was detected by immunohistochemistry. As compared with control group, the content of MDA, myocardial AI and the expression of Bcl-2, Bax proteins were increased significantly in I/R group, but the content of MDA, myocardial AI and the expression of Bax protein were decreased obviously and the expression of Bcl-2 protein was up-regulated in EP group (P<0.05). These results demonstrate that EP could inhibit apoptosis of cardiac myocytes possibly via alleviating oxidative stress, up-regulating Bcl-2 and down-regulating Bax proteins.