AIM： To investigate the role of oxidative stress in regulating the functional expression of P-glycoprotein（P-gp） in mitochondria of D407 cells.METHODS： D407 cells were exposed to different ranges of concentrations...AIM： To investigate the role of oxidative stress in regulating the functional expression of P-glycoprotein（P-gp） in mitochondria of D407 cells.METHODS： D407 cells were exposed to different ranges of concentrations of H2O2. The mitochondrial location of P-gp in the cells subjected to oxidative stress was detected by confocal analysis. Expression of P-gp in isolated mitochondria was assessed by Western blot. The pump activity of P-gp was evaluated by performing the efflux study on isolated mitochondria with Rhodamine 123（Rho-123） alone and in the presence of P-gp inhibitor（Tariquidar） using flow cytometry analysis. The cells were pretreated with 10 mmol/L N-acetylcysteine（NAC） for 30 min before exposing to H2O2, and analyzed the mitochondrial extracts by Western blot and flow cytometry.RESULTS： P-gp was co-localized in the mitochondria by confocal laser scanning microscopy, and it was also detected in the mitochondria of D407 cells using Western blot. Exposure to increasing concentrations of H2O2 led to gradually increased expression and location of P-gp in the mitochondria of cells. Rho-123 efflux assay showed higher uptake of Rho-123 on isolated mitochondria in the presence of Tariquidar both in normal and oxidative stress state. H2O2 up-regulated P-gp in D407 cells, which could be reversed by NAC treatment. CONCLUSION： H2O2 could up-regulate the functional expression of P-gp in mitochondria of D407 cells, while antioxidants might suppress oxidative-stress-induced over-expression of functional P-gp. It is indicative that limiting the mitochondrial P-gp transport in retinal pigment epithelium cells would be to improve the effect of mitochondria-targeted antioxidant therapy in age-related macular degeneration-like retinopathy.展开更多
Objective：To investigate the effects of Heijiangdan Ointment（黑绛丹膏,HJD） on oxidative stress in （60）Co γ-ray radiation-induced dermatitis in mice.Methods：Female Wistar mice with grade 4 radiation dermatitis i...Objective：To investigate the effects of Heijiangdan Ointment（黑绛丹膏,HJD） on oxidative stress in （60）Co γ-ray radiation-induced dermatitis in mice.Methods：Female Wistar mice with grade 4 radiation dermatitis induced by ^（60）Co γ-rays were randomly divided into four groups（n=12 per group）;the HJD-treated,recombinant human epidermal growth factor（rhEGF）-treated,Trolox-treated,and untreated groups,along with a negative control group.On the 11 th and 21 st days after treatment,6 mice in each group were chosen for evaluation.The levels of superoxide dismutase（SOD）,malondialdehyde（MDA）,and lactate dehydrogenase（LDH） were detected using spectrophotometric methods.The fibroblast mitochondria were observed by transmission electron microscopy（TEM）.The expressions of fibroblast growth factor 2（FGF-2） and transforming growth factor β1（TGF-β1） were analyzed by western blot.Results：Compared with the untreated group,the levels of SOD,MDA and LDH,on the 11 th and 21 st days after treatment showed significant difference（P〈0.05）.TEM analysis indicated that fibroblast mitochondria in the untreated group exhibited swelling and the cristae appeared fractured,while in the HJD group,the swelling of mitochondria was limited and the rough endoplasmic reticulum appeared more relaxed.The expressions of FGF-2 and TGF-β1 increased in the untreated group compared with the negative control group（P〈0.05）.After treatment,the expression of FGF-2,rhEGF and Trolox in the HJD group were significantly increased compared with the untreated group（P〈0.05）,or compared with the negative control group（P〈0.05）.The expression of TGF-β1 showed significant difference between untreated and negative control groups（P〈0.05）.HJD and Trolox increased the level of TGF-β1 and the difference was marked as compared with the untreated and negative control groups（P〈0.05）.Conclusion：HJD relieves oxidative stress-induced injury,increases the antioxidant activity,mitigates the fibroblast mitochondrial damage,up-regulates the expression of growth factor,and promotes mitochondrial repair in mice.展开更多
Long term suppression of succinate dehydrogenase by selective inhibitor 3-nitropropionic acid has been used in rodents to model Huntington＇s disease where mitochondrial dysfunction and oxidative damages are primary p...Long term suppression of succinate dehydrogenase by selective inhibitor 3-nitropropionic acid has been used in rodents to model Huntington＇s disease where mitochondrial dysfunction and oxidative damages are primary pathological hallmarks for neuronal damage. Improvements in learning and memory abilities, recovery of energy levels, and reduction of excitotoxicity damage can be achieved through activation of Adenyl cyclase enzyme by a specific phytochemical forskolin. In this study, intraperitoneal administration of 10 mg/kg 3-nitropropionic acid for 15 days in rats notably reduced body weight, worsened motor cocordination（grip strength, beam crossing task, locomotor activity）, resulted in learning and memory deficits, greatly increased acetylcholinesterase, lactate dehydrogenase, nitrite, and malondialdehyde levels, obviously decreased adenosine triphosphate, succinate dehydrogenase, superoxide dismutase, catalase, and reduced glutathione levels in the striatum, cortex and hippocampus. Intragastric administration of forskolin at 10, 20, 30 mg/kg dose-dependently reversed these behavioral, biochemical and pathological changes caused by 3-nitropropionic acid. These results suggest that forskolin exhibits neuroprotective effects on 3-nitropropionic acid-induced Huntington＇s disease-like neurodegeneration.展开更多
Objective： To investigate the cytoprotective effects of Saeng-kankunbi-tang（生肝健脾汤, SKT）, a herbal prescription consisting of Artemisia capillaris and Alisma canaliculatum, and its underlying mechanism involved...Objective： To investigate the cytoprotective effects of Saeng-kankunbi-tang（生肝健脾汤, SKT）, a herbal prescription consisting of Artemisia capillaris and Alisma canaliculatum, and its underlying mechanism involved. Methods： In mice, blood biochemistry and histopathology were assessed in carbon tetrachloride（CCl4）-induced oxidative hepatic injury in vivo. The animal groups included vehicle-treated control, CCl4, SKT 500 mg/（kg·day） CCl4＋SKT 200 or 500 mg/（kg·day）. In Hep G2 cell, tert-butyl hydroperoxide（t BHP） induced severe oxidative stress and mitochondrial dysfunction in vitro. The cyto-protective effects of SKT were determined by 3-（4,5-dimethylthiazol）-2,5-diphenyltetrazolium bromide（MTT） assay, fluorescence activated cell sorting analysis and western blotting. Results： The administration of SKT prevented liver damage induced by CCl4 in mice, by inhibition of hepatocyte degeneration and inflammatory cell infiltration as well as plasma parameters such as alanine aminotransferase（P〈0.01）. Moreover, treatment with t BHP induced hepatocyte death and cellular reactive oxygen species production in hepatocyte cell line. However, SKT pretreatment（30–300 μg/m L） reduced this cell death and oxidative stress（P〈0.01）. More importantly, SKT inhibited the ability of t BHP to induce changes in mitochondrial membrane transition in cell stained with rhodamine 123（P〈0.01）. Furthermore, treatment with SKT induced extracellular signal-regulated kinases-mediated nuclear factor erythroid-2-related factor 2（Nrf2） activation as well as the expressions of heme oxygenase 1 and glutamate-cystein ligase catalytic, Nrf2 target genes. Conclusion： SKT has the ability to protect hepatocyte against oxidative stress and mitochondrial damage mediated by Nrf2 activation.展开更多
基金Supported by National Natural Science Foundation of China(No.81400424)Guangdong Medical Science Foundation(No.A2015151)
文摘AIM： To investigate the role of oxidative stress in regulating the functional expression of P-glycoprotein（P-gp） in mitochondria of D407 cells.METHODS： D407 cells were exposed to different ranges of concentrations of H2O2. The mitochondrial location of P-gp in the cells subjected to oxidative stress was detected by confocal analysis. Expression of P-gp in isolated mitochondria was assessed by Western blot. The pump activity of P-gp was evaluated by performing the efflux study on isolated mitochondria with Rhodamine 123（Rho-123） alone and in the presence of P-gp inhibitor（Tariquidar） using flow cytometry analysis. The cells were pretreated with 10 mmol/L N-acetylcysteine（NAC） for 30 min before exposing to H2O2, and analyzed the mitochondrial extracts by Western blot and flow cytometry.RESULTS： P-gp was co-localized in the mitochondria by confocal laser scanning microscopy, and it was also detected in the mitochondria of D407 cells using Western blot. Exposure to increasing concentrations of H2O2 led to gradually increased expression and location of P-gp in the mitochondria of cells. Rho-123 efflux assay showed higher uptake of Rho-123 on isolated mitochondria in the presence of Tariquidar both in normal and oxidative stress state. H2O2 up-regulated P-gp in D407 cells, which could be reversed by NAC treatment. CONCLUSION： H2O2 could up-regulate the functional expression of P-gp in mitochondria of D407 cells, while antioxidants might suppress oxidative-stress-induced over-expression of functional P-gp. It is indicative that limiting the mitochondrial P-gp transport in retinal pigment epithelium cells would be to improve the effect of mitochondria-targeted antioxidant therapy in age-related macular degeneration-like retinopathy.
基金Supported by the National Natural Science Foundation of China(No.30973745)
文摘Objective：To investigate the effects of Heijiangdan Ointment（黑绛丹膏,HJD） on oxidative stress in （60）Co γ-ray radiation-induced dermatitis in mice.Methods：Female Wistar mice with grade 4 radiation dermatitis induced by ^（60）Co γ-rays were randomly divided into four groups（n=12 per group）;the HJD-treated,recombinant human epidermal growth factor（rhEGF）-treated,Trolox-treated,and untreated groups,along with a negative control group.On the 11 th and 21 st days after treatment,6 mice in each group were chosen for evaluation.The levels of superoxide dismutase（SOD）,malondialdehyde（MDA）,and lactate dehydrogenase（LDH） were detected using spectrophotometric methods.The fibroblast mitochondria were observed by transmission electron microscopy（TEM）.The expressions of fibroblast growth factor 2（FGF-2） and transforming growth factor β1（TGF-β1） were analyzed by western blot.Results：Compared with the untreated group,the levels of SOD,MDA and LDH,on the 11 th and 21 st days after treatment showed significant difference（P〈0.05）.TEM analysis indicated that fibroblast mitochondria in the untreated group exhibited swelling and the cristae appeared fractured,while in the HJD group,the swelling of mitochondria was limited and the rough endoplasmic reticulum appeared more relaxed.The expressions of FGF-2 and TGF-β1 increased in the untreated group compared with the negative control group（P〈0.05）.After treatment,the expression of FGF-2,rhEGF and Trolox in the HJD group were significantly increased compared with the untreated group（P〈0.05）,or compared with the negative control group（P〈0.05）.The expression of TGF-β1 showed significant difference between untreated and negative control groups（P〈0.05）.HJD and Trolox increased the level of TGF-β1 and the difference was marked as compared with the untreated and negative control groups（P〈0.05）.Conclusion：HJD relieves oxidative stress-induced injury,increases the antioxidant activity,mitigates the fibroblast mitochondrial damage,up-regulates the expression of growth factor,and promotes mitochondrial repair in mice.
文摘Long term suppression of succinate dehydrogenase by selective inhibitor 3-nitropropionic acid has been used in rodents to model Huntington＇s disease where mitochondrial dysfunction and oxidative damages are primary pathological hallmarks for neuronal damage. Improvements in learning and memory abilities, recovery of energy levels, and reduction of excitotoxicity damage can be achieved through activation of Adenyl cyclase enzyme by a specific phytochemical forskolin. In this study, intraperitoneal administration of 10 mg/kg 3-nitropropionic acid for 15 days in rats notably reduced body weight, worsened motor cocordination（grip strength, beam crossing task, locomotor activity）, resulted in learning and memory deficits, greatly increased acetylcholinesterase, lactate dehydrogenase, nitrite, and malondialdehyde levels, obviously decreased adenosine triphosphate, succinate dehydrogenase, superoxide dismutase, catalase, and reduced glutathione levels in the striatum, cortex and hippocampus. Intragastric administration of forskolin at 10, 20, 30 mg/kg dose-dependently reversed these behavioral, biochemical and pathological changes caused by 3-nitropropionic acid. These results suggest that forskolin exhibits neuroprotective effects on 3-nitropropionic acid-induced Huntington＇s disease-like neurodegeneration.
基金Supported by the National Research Foundation of Korea Grant funded by the Korea government(No.2014R1A2A2A01007375,No.2012R1A5A2A42671316)
文摘Objective： To investigate the cytoprotective effects of Saeng-kankunbi-tang（生肝健脾汤, SKT）, a herbal prescription consisting of Artemisia capillaris and Alisma canaliculatum, and its underlying mechanism involved. Methods： In mice, blood biochemistry and histopathology were assessed in carbon tetrachloride（CCl4）-induced oxidative hepatic injury in vivo. The animal groups included vehicle-treated control, CCl4, SKT 500 mg/（kg·day） CCl4＋SKT 200 or 500 mg/（kg·day）. In Hep G2 cell, tert-butyl hydroperoxide（t BHP） induced severe oxidative stress and mitochondrial dysfunction in vitro. The cyto-protective effects of SKT were determined by 3-（4,5-dimethylthiazol）-2,5-diphenyltetrazolium bromide（MTT） assay, fluorescence activated cell sorting analysis and western blotting. Results： The administration of SKT prevented liver damage induced by CCl4 in mice, by inhibition of hepatocyte degeneration and inflammatory cell infiltration as well as plasma parameters such as alanine aminotransferase（P〈0.01）. Moreover, treatment with t BHP induced hepatocyte death and cellular reactive oxygen species production in hepatocyte cell line. However, SKT pretreatment（30–300 μg/m L） reduced this cell death and oxidative stress（P〈0.01）. More importantly, SKT inhibited the ability of t BHP to induce changes in mitochondrial membrane transition in cell stained with rhodamine 123（P〈0.01）. Furthermore, treatment with SKT induced extracellular signal-regulated kinases-mediated nuclear factor erythroid-2-related factor 2（Nrf2） activation as well as the expressions of heme oxygenase 1 and glutamate-cystein ligase catalytic, Nrf2 target genes. Conclusion： SKT has the ability to protect hepatocyte against oxidative stress and mitochondrial damage mediated by Nrf2 activation.
