4-hydroxy-2-nonenal as a marker of the oxidative stress in brains of dogs with canine distemper / 4-hidroxi-2-nonenal como marcador de estresse oxidativo no cérebro de cães com cinomose

Canine Distemper is a disease caused by Canine morbillivirus (CM), a pantropic virus that can affect the central nervous system (CNS), causing demyelination. However, the pathogenesis of this lesion remains to be clarified. Brain samples of 14 naturally infected dogs by CM were analyzed to evaluate the presence of oxidative stress and demyelination. RT-PCR assay was performed to confirm a diagnosis of canine distemper in the brain, immunohistochemistry anti-CM was used to localize the viral proteins in the tissue, and anti-4-hydroxy-2-nonenal (4-HNE) was a marker of a product of lipid peroxidation. The results showed the presence of viral proteins in the demyelinated area with the presence of 4-HNE. Our results suggest that the CM virus infection causes oxidative stress leading to lipid peroxidation, which causes tissue damage and demyelination. In conclusion, oxidative stress plays a significant role in canine distemper pathogenesis in the CNS.(AU)

A cinomose canina é uma doença causada pelo Morbilivírus canino (CM), um vírus pantrópico que pode afetar o sistema nervoso central (SNC), causando desmielinização. No entanto, a patogênese dessa lesão não está totalmente esclarecida. RT-PCR e imuno-histoquímica foram realizadas para confirmação do diagnóstico de cinomose em amostras de encéfalo de 14 cães naturalmente infectados. Após confirmação, foi realizada uma avaliação do estresse oxidativo por imuno-histoquímica com uso de anti-4-hidroxi-nonenal (4HNE) como marcador de produtos resultantes da peroxidação lipídica. Os resultados sugerem que a infecção pelo CM causa estresse oxidativo no tecido, levando a peroxidação lipídica, a qual causa danos ao tecido, culminando com desmielinização. Conclui-se que o estresse oxidativo tem papel importante na patogênese da cinomose canina no sistema nervoso central.(AU)

The role of ALDH2 in tumorigenesis and tumor progression: Targeting ALDH2 as a potential cancer treatment

A major mitochondrial enzyme for protecting cells from acetaldehyde toxicity is aldehyde dehydrogenase 2 (ALDH2). The correlation between ALDH2 dysfunction and tumorigenesis/growth/metastasis has been widely reported. Either low or high ALDH2 expression contributes to tumor progression and varies among different tumor types. Furthermore, the ALDH2∗2 polymorphism (rs671) is the most common single nucleotide polymorphism (SNP) in Asia. Epidemiological studies associate ALDH2∗2 with tumorigenesis and progression. This study summarizes the essential functions and potential ALDH2 mechanisms in the occurrence, progression, and treatment of tumors in various types of cancer. Our study indicates that ALDH2 is a potential therapeutic target for cancer therapy.

Cytokeratin 13 promotes the radio-sensitivity of nasopharyngeal carcinoma HNE1 cells by inhibiting the PI3K/AKT/mTOR pathway via PTEN / 中国肿瘤生物治疗杂志

@#[Abstract] Objective: To investigate the effect of cytokeratin 13 (CK13) on radio-sensitivity of human nasopharyngeal carcinoma HNE1 cell line and its mechanism. Methods: HNE1 cells were divided into control group, anti-CK13#a group (CK13 knockdown), anti-CK13#b group (CK13 knockdown), control+sirolimus group (100 nmol/L sirolimus treatment for 1 h), and anti-CK13#a + sirolimus group (100 nmol/L sirolimus treatment for 1 h). After irradiation treatment (200 cGy/min irradiation for 5 min), cell proliferation in each group was measured by CCK-8 assay. Cell apoptosis rate in each group was determined by Flow cytometry. Expression of PI3K/AKT/mTOR signaling pathway related PTEN gene was detected by qPCR, and WB was used to detect the expressions of PI3K/AKT/mTOR signaling pathway related proteins. Results: In the case of radiotherapy, as compared with the control group, the proliferation of HNE1 cells after CK13 knockdown was significantly enhanced (P<0.01) while the apoptosis rate was significantly reduced (P<0.01), the contents of caspase-3 and γH2AX as well as the protein lever of PTEN in cells were significantly decreased, while the expressions of p-AKT and p-S6K were significantly increased (all P<0.01). Interestingly, additional treatment with sirolimus (PI3K/AKT/mTOR signaling pathway inhibitor) could rescue the accelerated cell proliferation and decreased cell apoptosis caused by CK13 knockdown (all P<0.05). Conclusion: CK13 knockdown can enhance the activity of PI3K/AKT/mTOR signaling pathway by down-regulating PTEN, and ultimately reduce the radio-sensitivity of nasopharyngeal carcinoma HNE1 cells.

Matrine attenuates oxidative stress and cardiomyocyte apoptosis in doxorubicin-induced cardiotoxicity maintaining AMPK/UCP2 pathway

Oxidative stress and cardiomyocyte apoptosis are involved in the pathogenesis of doxorubicin (DOX)-induced cardiotoxicity. Matrine is well-known for its powerful anti-oxidant and anti-apoptotic capacities. Our present study aimed to investigate the effect of matrine on DOX-induced cardiotoxicity and try to unearth the underlying mechanisms. Mice were exposed with DOX to generate DOX-induced cardiotoxicity or normal saline as control. H9C2 cells were used to verify the effect of matrine . DOX injection triggered increased generation of reactive oxygen species (ROS) and excessive cardiomyocyte apoptosis, which were significantly mitigated by matrine. Mechanistically, we found that matrine ameliorated DOX-induced uncoupling protein 2 (UCP2) downregulation, and UCP2 inhibition by genipin could blunt the protective effect of matrine on DOX-induced oxidative stress and cardiomyocyte apoptosis. Besides, 5'-AMP-activated protein kinase 2 () deficiency inhibited matrine-mediated UCP2 preservation and abolished the beneficial effect of matrine in mice. Besides, we observed that matrine incubation alleviated DOX-induced H9C2 cells apoptosis and oxidative stress level activating AMPK/UCP2, which were blunted by either AMPK or UCP2 inhibition with genetic or pharmacological methods. Matrine attenuated oxidative stress and cardiomyocyte apoptosis in DOX-induced cardiotoxicity maintaining AMPK/UCP2 pathway, and it might be a promising therapeutic agent for the treatment of DOX-induced cardiotoxicity.

Mechanisms of tanshinone Ⅱ_A in reducing 4-HNE-induced hepatocyte damage by activating PPARα / 中国中药杂志

Tanshinone Ⅱ_A( Tan Ⅱ_A),the liposoluble constituents of Salvia miltiorrhiza,can not only ameliorate the lipidic metabolism and decrease the concentration of lipid peroxidation,but also resist oxidation damage,scavenge free radicals and control inflammation,with a protective effect on prognosis after liver function impairment. Therefore,the studies on the exact mechanism of Tan Ⅱ_A in protecting the liver can provide important theoretical and experimental basis for the prevention and treatment effect of Tan Ⅱ_A for liver injury. In the present study,the protective effects and mechanism of Tan Ⅱ_A on 4-hydroxynonenal( 4-HNE)-induced liver injury were investigated in vitro. Normal liver tissues NCTC 1469 cells were used to induce hepatocytes oxidative damages by 4-HNE treatment. The protective effect of Tan Ⅱ_A on hepatocytes oxidative damages was detected by release amount of lactate dehydrogenase( LDH) analysis and hoechst staining. The protein expression changes of peroxisome proliferator-activated receptor α( PPARα) and peroxisome proliferator response element( PPRE) were analyzed by Western blot analysis in NCTC 1469 cells before and after Tan Ⅱ_A treatment. The gene expression changes of fatty aldehyde dehydrogenase( FALDH) were analyzed by Real-time polymerase chain reaction( PCR) analysis. The results showed that 4-HNE increased the release amount of LDH,lowered the cell viability of NCTC 1469 cells,and Tan Ⅱ_A reversed 4-HNE-induced hepatocyte damage. Western blot analysis and RT-PCR analysis results showed that 4-HNE decreased the expression of PPARα and FALDH and increased the expression of 4-HNE. However,the expression of PPARα and FALDH were increased significantly and the expression of 4-HNE was decreased obviously after Tan Ⅱ_A treatment. This study confirmed that the curative effect of Tan Ⅱ_A was obvious on hepatocytes damage,and the mechanism may be associated with activating PPARα and FALDH expression as well as scavenging 4-HNE.

Effects of Shenqi Fuzheng Injection on function and dynamics of skeletal muscle mitochondria in mice after colon cancer-cachexia / 中草药

Objective: To study the changes of mitochondria function and dynamics in colon cancer-cachexia mice and the effects of Shenqi Fuzheng Injection on these changes. Methods: A total of 40 female BLAB/c nu mice were randomly divided into control group, model group, low dose and high does of Shenqi Fuzheng Injection group, ten mice in every group. Except the control group, the mice of other groups were intraperitoneal injected with the Lovo cell line to establish the model of abdominal metastasis of colon cancer, then induced cancer cachexia. Marasmus and weight change were monitored the status of cancer cachexia in all groups. Shenqi Fuzheng Injection groups received intraperitoneal injection with 1.5 mL (2 times dose) and 3 mL (4 times dose) of drugs for every three days, seven consecutive times. After 21 days treatment, the mitochondrial related protein PGC-1, 4HNE and VDAC1 in the skeletal muscle were measured. The levels of adenosine triphosphate (ATP) and malondialdehyde (MDA) in the skeletal were detected. Results: Compared to the control group, the mice in the model group and Shenqi Fuzheng group suffered from cancer cachexia. Compared with the control group, the level of ATP in the skeletal muscle was lower in model group; The protein expression level of PGC-1 and 4HNE were remarkably increased (P 0.05); The level of MDA was significantly increased (P 0.05); The level of MDA was lower (P < 0.05). Conclusion: Shenqi Fuzheng Injection can significantly improve the status of colon cancer cachexia in mice, which may be related to the improvement of the mitochondrial function and the relieving of the mitochondrial oxidative damage.

Oxidative stress and sex difference in liver of rats exposed to cyclohexanone / Estrés oxidativo y diferencia sexual en el hígado de ratas expuestas a ciclohexanona

Cyclohexanone is widely used in industry for the organic synthesis of chemicals such as adipic acid, caprolactam, polyvinyl chloride and its copolymers, and methacrylate ester polymers. Its mechanism of toxicity, especially oxidative stress, is rarely reported in cyclohexanone toxicity studies. In this study, we evaluate oxidative stress immunohistochemically in the livers of rats exposed to cyclohexanone. Rats were exposed to 0 ppm and 625 ppm cyclohexanone for 6 h/day, 5 days/week, for 13 weeks via whole-body inhalation. All rats were sacrificed at the end of exposure and livers were removed and prepared for histological examination. Histopathology indicated an increase in bile duct hyperplasia in the liver was only observed in the cyclohexanone-exposed group, compared to that in the control group in males. Immunohistochemistry showed 4-HNE immunoreactivity in the cytoplasm of hepatocytes in the liver. Immunoreactivity was significantly stronger in the cyclohexanone-exposed group compared to the control group in both sexes. However, it was significantly stronger in males compared to females. This result shows a sex-based difference in the expression of oxidative stress in response to cyclohexanone exposure.

La ciclohexanona se usa ampliamente en la industria para la síntesis orgánica de sustancias químicas, como el ácido adípico, la caprolactama, el cloruro de polivinilo y sus copolímeros y los polímeros del éster metacrilato. Su mecanismo de toxicidad, especialmente el estrés oxidativo, se observa raramente en los estudios de toxicidad de la ciclohexanona. En el presente estudio, evaluamos el estrés oxidativo a través de la inmunohistoquímica en hígados de ratas expuestas a la ciclohexanona. Las ratas fueron expuestas a 0 ppm y 625 ppm de ciclohexanona por 6 horas diarias, 5 días a la semana durante 13 semanas, mediante inhalación corporal total. Al final de la exposición, se sacrificaron las ratas y se extirparon sus hígados para el examen histológico. La histopatología indicó que se observó un aumento de la hiperplasia del conducto biliar solamente en el grupo expuesto a la ciclohexanona, en comparación con el grupo de control en machos. La inmunohistoquímica mostró una inmunorreactividad al 4-HNE en el citoplasma de los hepatocitos del hígado. La inmunorreactividad fue significativamente mayor en el grupo expuesto a la ciclohexanona, en comparación con el grupo control en ambos sexos. Sin embargo, fue significativamente mayor en los machos, en comparación con el hígado de las hembras. Este resultado muestra una diferencia basada en el sexo, en la expresión del estrés oxidativo en respuesta a la exposición a la ciclohexanona.

Effect of anti-human IgM antibody on biological characteristics of human nasopharyngeal carcinoma HNE-1 cell line in vitro and in vivo / 重庆医学

Objective To investigate the effect of anti-human immunoglobulin M (IgM) on proliferation,apoptosis,cell cycle and tumor formation in human nasopharyngeal carcinoma HNE-1 cell line in vitro and in vivo.Methods After treatment with anti-human IgM antibody,proliferation of HNE-1 cells was observed by cell proliferation inhibition assay,apoptosis and cell cycle of HNE-1 cells were detected by flow cytometry,and apoptotic cells were detected by TUNEL staining.Nude mouse models were constructed,and were injected intraperitoneally with anti-human IgM antibodies (once every 3 days).The growth of transplanted tumor was observed once every 4 days.After the fifth injection,the expression levels of IgM and gp96 protein in transplanted tumor were observed by immunohistochemical method (streptavidin-peroxidase conjugated method,SP).Results MTS assay showed that anti-human IgM antibody can significantly inhibit the proliferation of HNE-1 cells in concentration-and time-dependent manner (P＜0.05).Flow cytometry showed that the anti-human IgM antibody promoted a significant decrease in percentage of cells in G1 phase,a significant increase in percentage of cells in S phase,and a significant increase in apoptotic rate of HNE-1 cells (P＜0.05).TUNEL staining showed that the anti-human IgM antibody promoted apoptosis of HNE-1 cells (P＜0.01).Transplantation tumor experiment showed that anti-human IgM antibody can significantly inhibit the volume and weight of transplanted tumor (P＜0.05).The immunohistochemistry showed that the expression levels of IgM and gp96 proteins in mouse transplanted tumors after intraperitoneal injection with anti-human IgM antibodies were significantly lower than those of the control group (P＜0.05).Conclusion The anti-human IgM anti-body could effectively inhibit the proliferation of HNE-1 cells,promote apoptosis,and arrest cell cycle.Anti-human IgM antibody could also inhibit the growth of transplanted tumor in nude mouse,which might be related to inhibition of the expressions of IgM and gp96 proteins.

Bifidobacteria relieve CPFX-induced testosterone reduction in mice / 基础医学与临床

Objective To explore if bacillus bifidus relieve CPFX-induced testosterone reduction in mouse testes.Methods Twenty-four male mices were divided into 4 groups, then administered saline for 6 days (Sal6 group), CPFX for 6 days (A6 group), CPFX for 6 days followed by bifidobacteria treatment for the next 6 days (A6+P6 group), CPFX for 6 days and then saline for the next 6 days (A6+Sal6 group).We detected serum levels of testosterone by RIA, as well as levels of steroidogenic enzymes mRNA [cholesterol side-chain cleavage enzyme (P450scc) and steroidogenic acute regulatory protein (StAR)] and NF-E2-related factor2 (Nrf2) mRNA in testes by real-time PCR, Nrf2, heme oxygenase-1 (HO-1), and 4-hydroxy-2-nonenal (4-HNE) by Western blot and4-HNE by Immunohistochemistry.Results The A6 group had significantly lower serum testosterone levels compared with the Sal6 group (P<0.001), the A6+P6 group had significantly higher compared with the A6 (P<0.001) and A6+Sal6 groups (P<0.01).The A6 group had significantly lower StAR mRNA compared with the Sal6 group (P<0.001), the A6+P6 group had significantly higher level compared with the A6 (P<0.01) and A6+Sal6 groups (P<0.01).The A6 group had significantly lower P450scc mRNA as compared with the Sal6 group (P<0.001), the A6+P6 group had significantly higher compared with the A6 (P<0.001) and A6+Sal6 groups (P<0.05).The A6 group had significantly lower Nrf2 compared with the Sal6 group (P<0.001), the A6+P6 group had significantly higher compared with the A6(P<0.01) and A6+Sal6 groups (P<0.05).The A6 group higher 4-HNE expression compared with the Sal6 group, the A6+P6 group had significantly lower compared with the A6 (P<0.01) and A6+Sal6 groups (P<0.05).Conclusions Bifidobacteria the reduction of CPFX-induced testosterone reduction, and these effects may potentially explained by Nrf2 inflammatory signaling pathway.

Modificações em proteínas induzidas por compostos eletrofílicos. possível papel em esclerose lateral amiotrófica / Modifications in proteins induced by electrophilic compounds. Possible role in amyotrophic lateral

Danos em biomoléculas podem ocorrer a partir de uma interação direta entre as biomoléculas e espécies reativas de oxigênio e nitrogênio como também, pela reação de produtos secundários dessas espécies como eletrófilos gerados na peroxidação lipídica. Alguns desses produtos secundários possuem estabilidade química maior que as espécies reativas das quais foram derivadas e podem se ligar covalentemente as biomoléculas comprometendo o funcionamento normal das mesmas. Portanto, modificações em proteínas por aldeídos gerados na lipoperoxidação têm sido investigadas por suas implicações com desordens patológicas relacionadas à agregação proteica, e modificações em diversas vias de sinalização amplificando os efeitos deletérios em sistemas biológicos. Os objetivos desse trabalho foi contribuir na elucidação dos mecanismos moleculares associados ao desenvolvimento da esclerose lateral amiotrófica (ELA) através da identificação, caracterização e quantificação de modificações póstraducionais em proteínas pelos aldeídos 4-hidroxi-2-hexenal (HHE) e trans-4-hidroxi-2-nonenal (HNE) in vitro (citocromo c) e in vivo (modelo ELA) a partir de técnicas de Western blot, imunoprecipitação e espectrometria de massa com abordagem proteômica de "shotgun" em ratosSOD1G93A modelo de esclerose lateral amiotrófica (ELA). Estudos com citocromo c mostraram a ligação dos aldeídos ao citocromo c e mecanismos de reação foram propostos. Foram encontrados seis peptídeos modificados por HHE e um para o HNE, e o peptídeo TGPNLHGLFGR se mostrou modificado pelos dois aldeídos paralelamente. Foi demonstrado que a histidina 33 é um "hot spot" frente as adições pelos aldeídos. Nas análises por western blot das proteínas ligadas a aldeídos foi possível observar uma tendência de aumento na concentração de proteínas ligadas ao HNE nos animais ELA, mais acentuada nas amostras de 70 dias comparadas ao controle. Com relação aos resultados obtidos com HHE tanto os animais pré-sintomáticos quanto os sintomáticos não apresentaram diferenças de HHE-proteína, tantonos controles quanto nos animais ELA. Nas amostras dos animais sintomáticos não detectamos diferença significativa na concentração de aldeído-proteína entre os grupos. Já as análises proteômicas revelaram 24 proteínas diferencialmente expressas, com destaque para proteínas com os maiores valores de significância (p-value), como a ubiquitina no grupo dos pré- sintomáticos e a neurogranina, no grupo dos animais sintomáticos e várias proteínas de metabolismo energético, de neurofilamentos, proteínas de processos redox e proteínas ligadas o metabolismo de cálcio (fundamentais na fisiopatologia em ELA). Algumas proteínas importantes foram encontradas com exclusividades nos grupos pré-sintomáticos e sintomáticos pelo diagrama de Venn. Com relação a proteínas modificadas pelos aldeídos, foram encontradas algumas relevantes como a proteína 2 de interação com a polimerase delta que foi modificada por HNE via adição de Michael encontrada nos animais ELA pré-sintomáticos e sintomáticos, a catalase que foi encontrada modificada por HNE via base de Schiff apenas nos ELA pré- sintomáticos, e a tiol redutase induzível por interferon gama no grupo dos animais ELA sintomáticos. Com relação a proteínas modificadas por HHE, foram encontradas a Janus quinase e proteína 3 de interação com microtúbulo, modificadas tanto por adição de Michael quanto via base de Schiff nos animais ELA sintomáticos. É interessante ressaltar que algumas modificações encontradas em proteínas não caracterizadas podem indicar proteínas novas ainda não descritas como modificadas por esses aldeídos. Os resultados mostram que algumas das proteínas modificadas por HNE e HHE encontradas neste trabalho, estão relacionadas ao estresse redox, vias metabólicas energéticas, proteínas envolvidas na resposta a danos oxidativos, e processos inflamatórios. Tais modificações ocorrem não só no modelo de neurodegeneração, mas foram previamente descritas em outros processos patológicos, como doença cardiovascular, lesão hepática por uso crônico de álcool

Damage to biomolecules can occur from a direct interaction between biomolecules and reactive of oxygen and nitrogen species as well as from the reaction of secondary products of these species as electrophiles generated in lipid peroxidation. Some of these by-products have greater chemical stability than the derived reactive species and can bind to biomolecules compromising their normal function. Therefore, protein modifications by aldehydes generated during lipoperoxidation have been investigated for their implications with pathological disorders related to protein aggregation and modifications in signaling pathways amplifying the deleterious effects in biological systems. The aim of this work was to contribute to the elucidation of the molecular mechanisms associated with the development of amyotrophic lateral sclerosis (ALS) through the identification, characterization and quantification of posttranslational modifications in proteins by 4-hydroxy-2-hexenal (HHE) and trans-4-hydroxy-2- nonenal (HNE) in vitro, cytochrome c, and in vivo, rat model (SOD1G93A) of amyotrophic lateral sclerosis (ALS), throught Western blot techniques, and mass spectrometry with shotgun proteomics approach. The results showed the binding of aldehydes to cytochrome c. Six peptides were modified by HHE and one by HNE. The peptide TGPNLHGLFGR was modified by the two aldehydes. Histidine 33 has been shown to be a hot spot against aldehydes additions. By western blot analysis of the aldehyde-bound proteins, it was possible to observe a tendency of increase in the concentration of HNE-bound proteins in the ALS animals, more pronounced in the samples of 70 days compared to control samples. Regarding the results obtained with HHE, both pre-symptomatic and symptomatic animals did not show HHE-protein differences, both in controls and in ALS animals. We did not detect a significant difference in the aldehyde-protein concentration between the groups in the samples of the symptomatic animals. Proteomic analysis revealed 24 differentially expressed proteins, with emphasis on proteins with thehighest values of significance (p-value), such as the ubiquitin in the pre-symptomatic group and neurogranin in the group of the symptomatic animals and several proteins of the energetic metabolism pathways, neurofilaments, proteins of redox processes and proteins linked to calcium metabolism (fundamental in the pathophysiology of ALS). Some important proteins were found exclusivity in the pre-symptomatic and symptomatic groups by the Venn diagram. With regard to aldehyde-modified proteins, some relevant ones such as Delta-2 polymerase interaction protein, that was modified by HNE via the addition of Michael found in presymptomatic and symptomatic ELA animals, catalase that was found to be modified by HNE via Schiff's base only in pre-symptomatic ALS, and gamma interferon-inducible thiol reductase in the group of symptomatic ALS animals. Janus kinase and microtubule interaction protein 3, were found to be modified by Michael addition and Schiff base pathway respectively in symptomatic ALS animals. It is interesting to note that some modifications found in uncharacterized proteins may indicate new proteins not yet described as modified by these aldehydes. The results show that some of the proteins modified by HNE and HHE found in this work are related to redox stress, energetic metabolic pathways, proteins involved in the response to oxidative damage, and inflammatory processes. Such modifications occur not only in the neurodegeneration model, but were previously described in other pathological processes, such as cardiovascular disease, liver injury due to chronic alcohol use

4-hydroxy-2(E)-Nonenal facilitates NMDA-Induced Neurotoxicity via Triggering Mitochondrial Permeability Transition Pore Opening and Mitochondrial Calcium Overload

N-methyl-D-aspartate (NMDA) receptor-mediated excitotoxicity is one of the major causes for neuronal cell death during cerebral ischemic insult. Previously, we reported that the final product of lipid membrane peroxidation 4-hydroxy-2E-nonenal (HNE) synergistically increased NMDA receptor-mediated excitotoxicity (J Neurochem., 2006). In this study, we investigated the mechanism involved in the synergistic neuronal cell death induced by co-treatment with HNE and NMDA. Although neither HNE (1 microM) nor NMDA (2 microM) alone induced the death of cortical neurons, simultaneous treatment of neuronal cells with HNE and NMDA synergistically evoked the death of the cells. However, the synergistic effect on neuronal death was observed only in the presence of calcium. HNE neither increased the cytosolic calcium level ([Ca2+]i) nor altered the NMDA-induced intracellular calcium influx. However, HNE together with NMDA elevated the mitochondrial calcium level and depolarized the mitochondrial transmembrane potential. Furthermore, HNE evoked damage of isolated mitochondria at the cytosolic calcium level (200 nM), which is maximally induced by 2 microM NMDA. Consistently, ATP was depleted in neurons when treated with both HNE and NMDA together. Ciclopirox, a potent inhibitor of mitochondrial permeability transition pore opening (Br. J. Pharmacol., 2005), largely prevented the synergistic damage of mitochondria and death of cortical neurons. Therefore, although low concentrations of HNE and NMDA cannot individually induce neuronal cell death, they can evoke the neuronal cell death by synergistically accelerating mitochondrial dysfunction.

Anti-atherosclerotic Effect of Green Tea in Poluynsaturated Fatty Acids-treated Apo E KO Mice

Dietary fatty acids are under intense research to identify anti-atherogenic mechanisms, so we investigated green tea powder (GT) as a protector against atherogenesis originating from lipid peroxidation such as 4-hydroxynonemal (4-HNE) and malondialdehyde (MDA) in different dietary fatty acid-treated apo E KO mice. Growth rate and dietary efficiency were lower in apo E KO mice with or without LA compared to wild type. Plasma total cholesterol (TC) and triacylglycerol (TG) did not correspond to values in other tissues, but TG in heart tissue decreased significantly by GT after linoleic acid (LA) or docosahexaenoic acid (DHA) was administered. LA induced apoptosis as evidenced by changes in aorta morphology and immunohistochemistry. Lipid peroxides (LPO) was increased in apo E KO mice with or without LA corresponding to the accumulation of 4-HNE or MDA in the proximal aorta above the atria. GT consumption tended to reduce the primary causal mechanism of atherogenic phenomena such as oxidizability in both LA and DHA treated atherogenic mice. A high polyunsaturated fatty acids (PUFA) diet involved the changes on stress-induced apoptotic signaling by increasing caspase 3, cytochrome c, and nuclear factor-kappaB in the heart tissue, but decreasing the bcl-2 protein. However, GT remarkably reduced the expression of apoptotic signaling, in contrast to the PUFA diet. Therefore, the potential of GT as an anti-atherosclerotic dietary antioxidant was tested in this study.

Cilostazol Attenuates 4-hydroxynonenal-enhanced CD36 Expression on Murine Macrophages via Inhibition of NADPH Oxidase-derived Reactive Oxygen Species Production

Although anti-atherogenic effects of cilostazol have been suggested, its effects on the expression of SR in macrophages are unclear. This study investigated the role of cilostazol on CD36 expression of murine macrophages enhanced by HNE, a byproduct of lipid peroxidation. The stimulation of macrophages with HNE led to an increased expression of CD36, which was significantly attenuated by NAC, an antioxidant. Moreover, the increased production of ROS by HNE was completely abolished by NADPH oxidase inhibitors, DPI and apocynin, as well as by the 5-LO inhibitor, MK886, but not by inhibitors for other oxidases. This suggested that NADPH-oxidase and 5-LO were major sources of ROS induced by HNE. In addition, HNE-enhanced expression of CD36 was reduced by these inhibitors, which indicated a role for NADPH oxidase and 5-LO on CD36 expression. In our present study, cilostazol was a significant inhibitor of ROS production, as well as CD36 expression induced by HNE. An increase in NADPH oxidase activity by HNE was significantly attenuated by cilostazol, however cilostazol had no effect on HNE-enhanced 5-LO activity. Together, these results suggest that cilostazol attenuates HNE-enhanced CD36 expression on murine macrophages thorough inhibition of NADPH oxidase-derived ROS generation.

Reversal of Multidrug Resistance in NPC HNE-1(200) Cell Line by Oxymatrine / 中国药房

OBJECTIVE:To study the reversal mechanism of oxymatrine on multidrug resistance in the NPC HNE-1(200) cell line. METHODS: The multidrug resistant HNE-1(200) subline was derived from an HNE-1 cell line after exposure to fractionated X-irradiation. The expression of P-gp was tested by immunocytochemical method and western blot and that of mdr1 mRNA was measured by RT-PCR. RESULTS: After 24-hour treatment with oxymatrine, both the immunocytochemical method and western blot showed a decrease in the expression of P-gp. However, RT-PCR measurement showed that the expression of mdr1 mRNA of HNE-1 and HNE-1(200) was nonsignficant. CONCLUSION: Oxymatrine can reverse the multidrug resistance of NPC HNE-1(200) cell line to some degree by down-regulating the expression of P-gp. However, this medicine did no effect on the expression of mdr1 mRNA, suggesting the up-regulation of the expression of P-gp is under the control of multiple mechanism.

Effects of Trail combined with chenotherapeutics on proliferation and apoptosis of HNE-1 cell line / 重庆医科大学学报

Objective:To observe the effects of different concentrations of TRAIL and chemotherapeutic drugs(5-FU,DDP) on proliferation and apoptosis of HNE-1 cell lines ,both singlely and jointly.Methods:MTT was used to detect the inhibition rate.FCM was used to detect the apoptosis rate of cell.Results:The inhibition rate and apoptosis rate of combination use of TRAIL and 5-FU,DDP were significantly increased,compared with those of single use of TRAIL.Conclusion:The inhibition of proliferation and apoptosis of HNE-1 cell line can be induced by TRAIL,and the effects were higher by TRAIL combined with 5-FU and DDP.

Inhibitory Effect of Tea Polyphenols in Combination with Anti-tumor Drugs on Cell Proliferation of the Multiresistant HNE-1 Cell Line / 中国药房

OBJECTIVE: To study the joint action of tea polyphenols and common anti-tumor drugs(vincristine,pingyangmycin,5-Fu,cisplatin) in inhibiting cell proliferation of the multiresistant HNE-1(200) cell line in patients with nasopharyngeal carcinoma after radiographic exposure.METHODS: The low toxic dosage of tea polyphenols and 4 kinds of anti-tumor drugs were optimized by MTT method.Then the inhibition ratio of multiresistant HNE-1(200) cell line treated by tea polyphenols and anti-tumor drugs waere detected.RESULTS: The low toxic dosage of tea polyphenols was 0.50 mg?mL-1.When the anti-tumor drugs were used in combination with tea polyphenols(0.50 mg?mL-1) as compared without,the inhibition ratio on HNE-1(200) cell line was increased by 100%～300%.CONCLUSIONS: Addition of tea polyphenols to anti-tumor drugs showed remarkable inhibitory effect on cell proliferation of HNE-1(200) cell line in patients with nasopharyngeal carcinoma,suggesting the reverse effect of tea polyphenols on resistant HNE-1(200) cell line.

Effects of oxymartine on expression of P-glycoprotein in HNE-1 cell line radiation therapy / 重庆医科大学学报

Objective:To investigate the effects of oxymartine on cell growth and the changes of P-glycoprotein in HNE-1 cell line after exposed to fractionated X-irradiation.Methods:By MTT,the cell proliferation was observed both before X-irradiation and after X-irradiation conditions with the extent of vincristine,oxymartine and verapamil respectively.P-glycoprotein expression was tested by immunohistochemistory method SP.Results:(1)Vincristine,oxymartine and verapamil were capable of inhibiting cell growth on HNE-1 cell line and fractionated X-irradiated HNE-1(200)cell subline;(2)Vincristine was less inhibitory on cell growth in HNE-1(200)cell subline than HNE-1 cell line,and the difference had statistic significance;(3)P-glycoprotein expression in HNE-1 cell line was negative while HNE-1(200)cell subline positive;oxymartine with low inhibitory concentration could increase the expression of P-glycoprotein in HNE-1(200) cell subline,which is similar to that of verapamil.Conclusion:(1)Oxymartine is capable of inhibiting cell growth on HNE-1 cell line and fractionated X-irradiated HNE-1(200)cell subline;(2)HNE-I(200)cell subline is resistant to vincristine;(3)Oxymartine with low inhibitory concentration could increase the expression of P-glycoprotein in HNE-1(200)cell subline.which is similar to that of verapamil.