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Objective:To investigate the effects of orlistat on the viability of human gall-bladder cancer (GBC) cells.Methods:The experimental study was conducted. The human GBC NOZ cells with high expression of FSAN was screened out through in vitro cultivating human GBC-SD, SGC-996 and NOZ cells. The cell proliferation assay, clone formation assay and protein detection experiment were used to analysis of the effects of orlistat on the viability of human GBC cells. Cell grouping: NOZ cells cultured with medium were set as the control group, cultured with medium + 10 μmol/L orlistat were set as the low-dose orlistat group, cultured with medium + 100 μmol/L orlistat were set as the high-dose orlistat group, respectively. Observation indicators: (1) expression of FASN protein in human GBC cells; (2) effects of orlistat on the proliferation of human GBC NOZ cells; (3) effects of orlistat on apoptosis of human GBC NOZ cells. Measurement data with normal distribution were represented as Mean± SD, the ANOVA test was used for comparison between groups and the least significant difference method was used for pairwise comparison. Results:(1) Expression of FASN protein in human GBC cells. Results of western blot showed that the relative expression of FASN protein in human GBC NOZ, GBC-SD and SGC-996 cells was 0.57±0.06, 0.12±0.04 and 0.10±0.02, respectively, showing a significant difference among them ( F=115.67, P<0.05). There were significant differences between the NOZ cells and the GBC-SD or the SGC-996 cells ( P<0.05), and there was no significant difference between the GBC-SD cells and the SGC-996 cells ( P>0.05). (2) Effects of orlistat on the proliferation of human GBC NOZ cells. ① Results of cell proliferation assay showed that the absorbance value of NOZ cells was 2.34±0.12, 1.57±0.08 and 1.07±0.13 in the control group, low-dose orlistat group and high-dose orlistat group, respectively, showing a significant difference among them ( F=205.88, P<0.05). ② Results of clone formation assay showed that the number of NOZ cells clones was 257±23, 153±11 and 83±11 in the control group, low-dose orlistat group and high-dose orlistat group, respectively, showing a significant difference among them ( F=92.64, P<0.05). ③Results of western blot showed that the relative expression of Cyclin-D1 protein of NOZ cells was 2.31±0.10, 1.52±0.05 and 1.23±0.11 in the control group, low-dose orlistat group and high-dose orlistat group, respectively, showing a significant difference among them ( F=120.73, P<0.05). The relative expression of CDK-4 protein of NOZ cells was 1.58±0.04, 1.21±0.02 and 1.19±0.04 in the control group, low-dose orlistat group and high-dose orlistat group, respectively, showing a signifi-cant difference among them ( F=110.45, P<0.05). (3) Effects of orlistat on apoptosis of human GBC NOZ cells. Results of western blot showed that the relative expression of Bcl-2 protein of NOZ cells was 1.07±0.03, 0.36±0.03 and 0.15±0.02 in the control group, low-dose orlistat group and high-dose orlistat group, respectively, showing a significant difference among them ( F=1 242.93, P<0.05). The relative expression of Bax protein of NOZ cells was 0.51±0.03, 0.38±0.05 and 1.38±0.04 in the control group, low-dose orlistat group and high-dose orlistat group, respectively, showing a signifi-cant difference among them ( F=583.51, P<0.05). Conclusion:Orlistat can inhibit the growth of human GBC NOZ cells and promote their apoptosis.
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OBJECTIVE： To study the effects of cimetidine on low dose rate irradiation-induced liver cell apoptosis in Beagle dogs. METHODS： Healthy male Beagle dogs were randomly divided into normal control group， model control group， positive drug group （lentinan， 21.33 mg/kg） and cimetidine low-dose， medium-dose and high-dose groups （5.33， 10.67， 21.33 mg/kg）， with 4 Beagle dogs each. Except for normal control group， other groups were given 60Co-γ accumulative irradiation （dosage rate： 0.040 8 mGy/min） for 23 d； the medication groups were given relevant medicine orally before irradiation， once a day. Twenty-four hours after stopping irradiation， TUNEL method was used to detect the apoptosis of liver cells in Beagle dogs. The percentage of apoptotic cells was calculated. The expression level of apoptosis-related proteins （Bax， Bcl-2， Caspase-3， p53） in liver tissue was detected by immunohistochemistry. RESULTS： Compared with normal control group， apoptotic cells and Bax， Caspase-3， p53 positive cells were increased significantly in liver tissue of Beagle dogs in model control group； the percentage of apoptotic cells， protein expression levels of Bax， Caspase-3 and p53 were increased significantly； Bcl-2 positive cells were decreased significantly， and its protein expression level was decreased significantly （P＜0.05 or P＜0.01）. Compared with model control group， above positive cells of liver tissue in Beagle dogs were changed to different extents in medication groups； the percentage of apoptotic cells and protein expression levels of p53 in medication groups， protein expression levels of Bax in positive drug group， cimetidine low-dose and high-dose groups as well as protein expression levels of Caspase-3 in cimetidine groups were decreased significantly； protein expression levels of Bcl-2 were increased significantly in cimetidine groups. The percentage of apoptotic cells in cimetidine medium-dose and high-dose groups as well as protein expression levels of Caspase-3 in cimetidine groups were all lower than positive control group. Protein expression level of p53 in cimetidine low-dose group was significantly higher than positive drug group （P＜0.05 or P＜0.01）. CONCLUSIONS： Cimetidine can inhibit the low dose rate irradiation-induced apoptosis of liver cells in Beagle dogs， and certainly protect liver cells against irradiation. The mechanism of it may be associated with up-regulating the protein expression of Bcl-2 and down-regulating the protein expression of Bax， Caspase-3 and p53 in liver cells.
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In this paper,the effects of active fractions of Ferula ferulaeoides on the growth and apoptosis of human gastric cancer cell MGC-803 transplantation tumor were systematically studied. The subcutaneous ectopic transplantation tumor model was established in human gastric cancer MGC-803 nude mice by cell suspension implantation method. The anti-tumor rate and organ index were used to evaluate the anti-tumor effect of the active fractions of F. ferulaeoides on the tumor-bearing nude mice. HE staining,TUNEL staining,RT-PCR,Western-blot and ELISA were used for pathological examination,apoptosis observation,and detection of apoptosis-related genes,proteins and cytokines expression. The results showed that as compared with the model group,the low,medium and high doses of the active fraction of F. ferulaeoides had inhibitory effects on xenografts in nude mice,respectively,in a dose-dependent manner; the apoptotic ratio was increased with the increase of drug concentration. As compared with the model group,F. ferulaeoides could down-regulate the expression of survivin mRNA in nude mice,and the protein expression levels of Bax,Bcl-2,caspase-3 and caspase-9 in tumor tissues of nude mice could be increased to different degrees in F. ferulaeoides groups. The contents of IL-10 and TGF-β1 in plasma of nude mice were decreased in high dose group of F. ferulaeoides active fractions. The results indicated that F. ferulaeoides can significantly inhibit the growth of human gastric cancer MGC-803 subcutaneously transplanted tumor,and its mechanism may be related with down-regulating the expression of survivin mRNA,and up-regulating the expression of apoptosis-related proteins Bax,caspase-3 and caspase-9.
Subject(s)
Animals , Humans , Mice , Apoptosis , Caspase 3 , Metabolism , Cell Line, Tumor , Cytokines , Metabolism , Ferula , Chemistry , Mice, Inbred BALB C , Mice, Nude , Neoplasm Transplantation , Plant Extracts , Pharmacology , Proto-Oncogene Proteins c-bcl-2 , Metabolism , Stomach Neoplasms , Drug Therapy , bcl-2-Associated X Protein , MetabolismABSTRACT
Objective:To investigate the effect of Schisandra chinensis polysaccharide(SCP)on the growth of brain tumor stem cells(BTSCs),and to clarify the mechanism of inhibiting the growth of BTSCs of SCP. Methods:The primary human glioma cells were cultured,then the BTSCs were isolated by CD133 immunomagnetic sorting.The neural stem cell surface markers CD133 and Nestin were detected by immunofluorescence assay.The proliferation rate of BTSCs was examined by MTT assay.Annexin V-PI analysis was used to analyze the apoptotic rate of BTSCs.The expression levels of Bax,Bcl-2 and Caspase-3 proteins in BTSCs in various groups were detected by ELISA assay.Results:The results of immunofluorescence staining showed that the expressions of CD133 and Nestin were positive in BTSCs.Compared with control group,the proliferation rates of BTSCs in 200,400 and 800 mg·L-1SCP groups were decreased,especially in 400 and 800 mg·L-1SCP groups(P<0.05).The results of Annexin V-PI analysis showed that the apoptotic rate of BTSCs in 800 mg·L-1SCP group was increased compared with control group(P<0.05).The ELISA results showed that the expression levels of Bax in 200,400 and 800 mg·L-1SCP groups were significantly increased(P<0.05),and the values of Bax/Bcl-2 were significantly increased(P<0.05);compared with control group,the Bcl-2 expression level in the BTSCs in 800 mg·L-1SCP group was decreased(P<0.05).The expression level of Caspase-3 protein in 800 mg·L-1SCP group was also significantly increased compared with control group(P<0.01).Conclusion:SCP could inhibit the growth of BTSCs,and the induction of apoptosis may be one of mechanisms.
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@#Objective To investigate the effect of Huangjiao granule on inflammatory factors and apoptosis-related proteins in rats with cerebral ischemia-reperfusion injury.Methods A total of 40 Sprague-Dawley rats were divided into sham operation group, model group and Huangjiao granule group, with ten rats in each group, and ten rats standby. The cerebral ischemia for two hours and reperfusion model was established by suture method. The sham operation group and the model group were given saline 10 ml/kg intragastrically 30 minutes before operation. The Huangjiao granule group was given Huangjiao granule solution 10 ml/kg (the content of crude drug was 1 g/ml) intragastrically 30 minutes before ischemia-reperfusion. Longa scoring method was used to evaluate the neurological function score 24 hours after reperfusion, while the percentage of cerebral infarction volume was detected by TTC staining, the pathological morphology of brain tissue was observed by HE staining, the cell apoptosis of brain tissue was detected with TUNEL, the levels of interleukin (IL)-1β, IL-6 and tumor necrosis factor α (TNF-α) in serum were detected by ELISA, the expression of cleaved-caspase 3, cleaved-caspase 9, Bax and Bcl-2 proteins of brain tissue was evaluated by Western blotting.Results Compared with the model group, the neurological score decreased, the percentage of cerebral infarction volume and the apoptosis rate of the brain tissue decreased in Huangjiao granule group (P<0.05). The levels of IL-1β, IL-6 and TNF-α in serum and the expression of cleaved-caspase 3, cleaved-caspase 9 and Bax proteins of brain tissues significantly decreased (P<0.05), and the expression of Bcl-2 protein of brain tissues inreased in Huangjiao granule group (P<0.05).Conclusion The protective effect of Huangjiao granule on rats with cerebral ischemia-reperfusion injury may be related to the inhibition of inflammatory response and the reduction of apoptosis.
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Objective: To investigate the effect of Schisandra chinensis polysaccharide (SCP) on the growth of brain tumor stem cells (BTSCs), and to clarify the mechanism of inhibiting the growth of BTSCs of SCP. Methods: The primary human glioma cells were cultured, then the BTSCs were isolated by CD133 immunomagnetic sorting. The neural stem cell surface markers CD133 and Nestin were detected by immunofluorescence assay. The proliferation rate of BTSCs was examined by MTT assay. Annexin V-PI analysis was used to analyze the apoptotic rate of BTSCs. The expression levels of Bax, Bcl-2 and Caspase-3 proteins in BTSCs in various groups were detected by ELISA assay. Results: The results of immunofluorescence staining showed that the expressions of CD133 and Nestin were positive in BTSCs. Compared with control group, the proliferation rates of BTSCs in 200, 400 and 800 mg · L-1 SCP groups were decreased, especially in 400 and 800 mg · L-1 SCP groups (P<0.05). The results of Annexin V-PI analysis showed that the apoptotic rate of BTSCs in 800 mg · L-1 SCP group was increased compared with control group (P<0.05). The ELISA results showed that the expression levels of Bax in 200, 400 and 800 mg · L-1 SCP groups were significantly increased (P<0.05), and the values of Bax/Bcl-2 were significantly increased (P<0.05); compared with control group, the Bcl-2 expression level in the BTSCs in 800 mg · L-1 SCP group was decreased (P<0.05). The expression level of Caspase-3 protein in 800 mg · L-1 SCP group was also significantly increased compared with control group (P<0.01). Conclusion: SCP could inhibit the growth of BTSCs, and the induction of apoptosis may be one of mechanisms.
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Objective: To investigate the regulatory effect of genipin (GP) on the gastric cancer SGC 7901 cells, and to clarify its mechanism. Methods: The SGC 7901 cells in logarithmic growth phase were selected and divided into control group and different concentrations (5. 0, 10. 0, and 20. 0 mg · L-1) of GP groups. MTT was used to detect the inhibitory rates of proliferation of SGC 7901 cells at different time points (24, 48, and 72 h). Transwell chamber cell invasion assay was used to detect the invasion ability of SGC 7901 cells in vitro. The expression levels of Bcl-2, Bax and caspase-3 proteins in the SGC 7901 cells were detected by Western blotting method. Results: The results of MTT assay showed that the inhibitory rates of proliferation of the cells in different concentrations of GP groups after treated with GP for different time were increased significantly compared with control group (P< 0. 01). The Transwell cell invasion assay results showed that compared with control group, the number of transmembrane cells in 10. 0 and 20. 0 mg · L-1 GP groups was decreased significantly after treated for 72 h (P< 0. 01). The results of Western blotting method showed that compared with control group, the expression level of Bcl-2 protein in 20. 0 mg · L-1 GP group was decreased after treated for 72 h (P<0. 01), and the expression levels of caspase-3 and Bax proteins were increased (P<0. 05). Conclusion; GP can inhibit the abilities of proliferation and invasion in vitro of SGC 7901 cells, and induce the apoptosis; its mechanism may be related to the regulation of Bcl-2, caspase-3, and Bax protein expressions.
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Objective:To investigate the effects of high thoracic epidural anesthesia (HTEA) on the cerebral blood flow (CBF) and hippocampal apoptosis-related proteins Bcl-2 and Bax during global cerebral ischemia and reperfusion (GCI) in rats.Methods:Fifteen-minute global ischemia was established by 4-vessel occlusion and epidural catheterization was performed through T4-5 intervertebral spaces in adult male Wistar rats.According to the different drugs infused into the epidural space,the rats were randomly divided into four groups:Sham group (0.9 ％ NaC1),Sham-HTEA group (0.25 ％ bupivacaine),GCI group (global cerebral ischemia,0.9 ％ NaC1) and HTEA group (global cerebral ischemia,0.25 ％ bupivacaine).And 0.25 ％bupivacaine or 0.9 ％ saline (20 μL·h-1) was infused continuously to the thoracic epidural space from 15 minutes before ischemia to 24 hours after reperfusion.Mean arterial pressure (MAP),heart rate (HR) and cerebral blood flow (CBF) were determined until 2 hours after reperfusion,and the hippocampal Bcl-2 and Bax proteins at 24 hours after reperfusion were examined by Western-blot.Results:Compared with the GCI group,HTEA group has no significant difference on MAP and HR during ischemia and 2 hours after reperfusion,andcompared with the Sham group,MAP in GCI group increased in ischemia 0 min and decreased in reperfusion 0 min.The CBF in HTEA group was significantly lower than that in GCI group (123.1％± 35.2％ vs 177.5％± 32.4％,P＜0.01) in reperfusion 10 min,and higher than that in GCI group during the hypoperfusion of 60 to 120 minutes after reperfusion (P＜0.05),and the ratio of Bax/Bcl-2 in hippocampus was significantly decreased in HTEA group 24 hours after reperfusion (P＜0.01).Conclusions:Continuous HTEA infusion of 0.25 ％ bupivacaine 20 μL ·h-1 could maintain the hemodynamic stability,and improve the CBF of hypoperfusion period in rats,as well as reduce the ratio of Bax/Bcl-2 at 24 hours after reperfusion.
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BACKGROUND: The dysfunction of the proteasome system has been implicated in neuronal degeneration. Apocynin, a specific inhibitor for nicotinamide adenine dinucleotide phosphate oxidase, has anti-inflammatory and anti-oxidant effects. However, the effect of apocynin on the neuronal cell death induced by proteasome inhibition has not been studied. METHODS: Using differentiated PC12 cells, in the respect of cell death process the suppressive effect of apocynin on the proteasome inhibition-mediated apoptosis was examined. RESULTS: The proteasome inhibitors MG132 and MG115 induced a decrease in Bid and Bcl-2 protein levels, an increase in Bax and p53 levels, mitochondrial depolarization, efflux of cytochrome c into cytosol and increase in caspases (-8, -9 and -3) activities. Treatment with apocynin attenuated the proteasome inhibitor-induced changes in the apoptosis-related protein levels, formation of reactive oxygen species, glutathione (GSH) depletion and cell death. CONCLUSIONS: Apocynin may attenuate the proteasome inhibitor-mediated apoptosis in differentiated PC12 cells by inhibiting the activation of the mitochondria-mediated pathway and the caspase-8- and Bid-dependent pathways. The preventive effect of apocynin appears to be attributed to inhibition of the production of reactive oxygen species and the depletion of cellular GSH contents.
Subject(s)
Animals , Antioxidants , Apoptosis , Caspases , Cell Death , Cytochromes c , Cytosol , Glutathione , NADP , Neurons , Oxidoreductases , PC12 Cells , Proteasome Endopeptidase Complex , Proteasome Inhibitors , Reactive Oxygen SpeciesABSTRACT
Objective:To evaluate the effects of periodontitis on the expression of apoptosis-related proteins in pancreas of rats with Type 2 Diabetes Mellitus.Methods:Spontaneously type 2 diabetic OLETF rats were randomly divided into 2 groups:diabetes with or without periodontitis(diabetes group and combination group).LETO rats with the same germline and the same age but having normal glucose tolerance were randomly divided into control group and periodontitis group.20 weeks after periodontitis were established,all the rats were sacrificed and the pancreas were pathologically examined by HE staining.The expression of Bax,Bcl-2 and Caspase-3 in the pancreas islet were detected by immunohistochemistry staining and semi-quantitative analysis.Results:The expression of Bax, Bcl-2 and Caspase-3 in the pancreas islet was no significant difference between control and periodontitis groups(P=0.324,P=0.091,P=0.852).Compared with diabetes group,the expression of Bax and Caspase-3 in combination group showed a significant increase(P=0.000,P=0.000),and the expression of Bcl-2 was significantly decreased(P=0.022).Conclusion:Under healthy conditions,periodontitis has no effect on the expression of apoptosis-related proteins in rat pancreas islet.However,in rats with diabe-tes,periodontitis may affect the expression of apoptosis-related proteins in pancreas islet.
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Objective To detect the apoptosis of femoral head cartilage cells and to observe the expression of apoptosis-related proteins in the tissues of the femoral head,as well as to explore the main pathway for regulating the apoptosis in steroid induced by juvenile rabbit models with avascular necrosis of femoral head.Methods The models with avascular necrosis of the femoral head and the control group model were made in New Zealand infancy albino rabbits induced by glucocorticoid(GC).The immunohistochemical method was used to detect the expressions of Caspase3,Caspase-8,apoptosis protease activating factor-1 (apaf-1),calpain-1 in the femoral heads.Results 1.The integrated optical density(IOD) values of Caspase-3 in GC-induced subgroup,the subgroup that was not induced by GC and control group were 25 142.72 ± 21 528.48,2 069.63 ± 1 096.96 and 301.80 ± 99.66,respectively.The IOD values of Caspase-8 in GC-induced subgroup,the subgroup respectively and the control group were 24 942.63 ± 18 942.99,2 016.31 ± 1 518.70,236.85 ±97.94,respectively.The IOD values of apaf-1 in GC-induced subgroup,the subgroup that was not induced by GC and the control group were 8 5 14.23 ± 6 384.20,1 118.49 ± 1 360.59,95.13 ± 38.05,respectively.The IOD values of calpain-1 in GC-induccd subgroup,the subgroup that was not induced by GC and control group were 9 636.71 ± 9 123.81,1 881.10 ± 3 277.86,126.71 ± 47.35,respectively.The IOD value differences of the Caspase-3 between GC-induced subgroup and the subgroup that was not induced by GC,the control group were extremely significant (H =l 1.470,23.996,P ＜ 0.01).The IOD value differences of the Caspase-8,apaf-1,calpain-1 between GC-induced subgroup and the control group were extremely significant (H =22.178,22.808,13.553,P ＜ 0.01).2.The linear regression analysis results showed that under the joint action of Caspase-8,apaf-1,calpain-1,only the Caspase-8 could significantly predict Caspase-3,and its regression equation regression got significant effect and could explain 40.3％ of the variance;while the regression effects of the apaf-1 and calpain-1 to Caspase-3 were not significant.Conclusion Death receptor pathway might play a major regulation role in the apoptotic process of avascular necrosis.
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BACKGROUND: 1-Methyl-4-phenylpyridinium (MPP+) causes a neuronal cell injury that is similar to the findings observed in Parkinson's disease. Caffeoylquinic acid derivatives have demonstrated anti-oxidant and anti-inflammatory effects. Nevertheless, the effect of 3,4,5-tricaffeoylquinic acid (3,4,5-triCQA) on the neuronal cell death due to exposure of parkinsonian toxin MPP+ remains unclear. METHODS: Using differentiated PC12 cells, the preventive effect of 3,4,5-triCQA on the MPP+-induced cell death in relation to apoptotic process was examined. RESULTS: MPP+ induced a decrease in Bid, Bcl-2 and survivin protein levels, increase in Bax levels, loss of the mitochondrial transmembrane potential, cytochrome c release, activation of caspases (-8, -9 and -3), cleavage of PARP-1, and an increase in the tumor suppressor p53 levels. 3,4,5-Tricaffeoylquinic acid attenuated the MPP+-induced changes in the apoptosis-related protein levels, formation of reactive oxygen species, depletion of GSH, nuclear damage and cell death. 3,4,5-Tricaffeoylquinic acid attenuated another parkinsonian neurotoxin rotenone-induced cell death. CONCLUSIONS: 3,4,5-Tricaffeoylquinic acid may attenuate the MPP+-induced apoptosis in PC12 cells by suppressing the activation of the mitochondrial pathway and the caspase-8- and Bid-dependent pathways. The preventive effect seems to be ascribed to its inhibitory effect on the formation of reactive oxygen species and depletion of GSH.
Subject(s)
Animals , 1-Methyl-4-phenylpyridinium , Apoptosis , Caspases , Cell Death , Cytochromes c , Membrane Potentials , Neurons , Parkinson Disease , PC12 Cells , Reactive Oxygen SpeciesABSTRACT
BACKGROUND: Flavonoid luteolin has been shown to exhibit cell protective effect. However, it is still uncertain whether the effect of luteolin on cellular toxicity of the parkinsonian toxin 6-hydroxydopamine is mediated by apoptosis-related protein activation. METHODS: In differentiated PC12 cells exposed to 6-hydroxydopamine in combination with luteolin, we observed the apoptosis-related protein activation, nuclear damage, formation of reactive oxygen species and cell death. RESULTS: 6-Hydroxydopamine caused apoptosis by inducing a decrease in Bid, Bcl-2, Bcl-xL and survivin levels, increase in Bax levels, cytochrome c release and activation of caspases. Treatment with luteolin reduced changes in the apoptosis-related protein levels, formation of reactive oxygen species, nuclear damage and cell death. CONCLUSIONS: Luteolin may reduce the 6-hydroxydopamine-induced apoptosis in differentiated PC12 cells by suppressing the activation of the caspase-8- and Bid-dependent pathways and the mitochondria-mediated apoptotic pathway, leading to caspase activation. The preventive effect of luteolin may be associated with its inhibitory effect on the production of reactive oxygen species. Luteolin may attenuate the oxidative stress and mitochondrial dysfunction-induced neuronal cell death take place in Parkinson's disease.
Subject(s)
Animals , Apoptosis , Caspases , Cell Death , Cytochromes c , Hypogonadism , Luteolin , Mitochondrial Diseases , Neurons , Ophthalmoplegia , Oxidative Stress , Oxidopamine , Parkinson Disease , PC12 Cells , Reactive Oxygen SpeciesABSTRACT
BACKGROUND: Defects in mitochondrial function have been shown to participate in the induction of neuronal cell injury. The extracellular-signal-regulated kinase (ERK) signaling pathway plays a crucial role in almost all cell functions, including proliferation, differentiation, survival, and death. However, the effect of ERK inhibition on oxysterol-induced apoptosis remains uncertain. METHODS: This study assessed the effect of ERK inhibition on the apoptotic effect of 7-ketocholesterol. RESULTS: Treatment with 7-ketocholesterol increased phosphorylated-ERK1/2 levels in differentiated PC12 cells, while the total amount of ERK was not altered. 7-Ketocholesterol decreased Bid and Bcl-2 levels, increased Bax and p53 levels, and promoted cytochrome c release, which elicits the activation of caspases (-8, -9, and -3), nuclear damage, and cell death. ERK and farnesyltransferase inhibitors inhibited the 7-ketocholesterol-induced phosphorylation of ERK1/2, activation of apoptosis-related proteins, and cell death in PC12 cells. CONCLUSIONS: The ERK and farnesyltransferase inhibitors, which did not exhibit toxicity, may inhibit the 7-ketocholesterol toxicity on differentiated PC12 cells by suppressing the activation of the caspase-8-dependent pathway as well as activation of the mitochondria-mediated cell-death pathway, leading to the activation of caspases. The inhibition of ERK may confer a beneficial protective effect against the neuronal cell injury induced by cholesterol oxidation products.
Subject(s)
Animals , Apoptosis , Caspases , Cell Death , Cholesterol , Cytochromes c , Farnesyltranstransferase , Ketocholesterols , Neurons , PC12 Cells , Phosphorylation , Phosphotransferases , ProteinsABSTRACT
Objective:To explore apoptosis changes of dilator muscles in the upper airway by detecting the expression of Bax, Bcl-2 in tensor veli palatini in patients with OSAHS.Method:The expression of Bax and Bcl-2 were detected in tensor veli palatini in 30 cases with OSAHS and 10 cases chronic tonsillitis without OSAHS by immunohistochemistry and image analytical system, and the results were analyzed.Result:①The expression levels of Bax in the OSAHS group increased significantly compared to control group(P<0.05), but there were no significant differences of Bcl-2 expression between two groups, the ratio of Bax/Bcl-2 increased significantly(P<0.05). ②There were positive correlations between AHI and the expression levels of Bax(r=0.697,P<0.01) respectively in the test group.Conclusion:The results indicate that apoptosis occurred in tensor veli palatini in patients with OSAHS, and the more severity of OSAHS , the more apoptosis.
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BACKGROUND: Protein casein kinase 2 is involved in signal transduction, cell growth, and apoptosis. However, it has not been elucidated whether parkinsonian toxin 1-methyl-4-phenylpyridinium (MPP+)-induced neuronal cell death is mediated by a casein-kinase-2-mediated pathway. METHODS: We monitored apoptosis-related protein activation, changes in the level of casein kinase 2, nuclear damage, and apoptosis in differentiated PC12 cells exposed to MPP+ in combination with casein kinase 2 inhibitor. RESULTS: Casein kinase 2 inhibitors [4,5,6,7-tetrabromobenzotriazole (TBB), 5,6-dichloro-1-beta-D-ribofuranosylbenzimidazole, and apigenin] reduced MPP+- and rotenone-induced cell death in differentiated PC12 cells. TBB inhibited the MPP+-induced activation of apoptosis-related proteins (decreases in Bid and Bcl-2 levels, increase in Bax levels, cytochrome c release, and caspase-3 activation), increase in casein kinase 2 levels, and nuclear damage. CONCLUSIONS: Administering casein kinase 2 inhibitor TBB at concentrations that do not induce toxic effects may reduce MPP+-induced cell death in differentiated PC12 cells by suppressing the apoptosis-related protein activation that leads to cytochrome c release and subsequent activation of caspase-3. The results suggest that MPP+-induced cell death process is mediated by a casein kinase 2 pathway.
Subject(s)
Animals , 1-Methyl-4-phenylpyridinium , Apoptosis , Casein Kinase II , Casein Kinases , Caseins , Caspase 3 , Cell Death , Cytochromes c , Neurons , PC12 Cells , Proteins , Signal Transduction , TriazolesABSTRACT
The changes of Fas.FasL and Bcl-2 expression in thyrocytes of patients with Graves'disease were investigated before and 2 weeks after 131Ⅰ administration. The results showed that 131Ⅰ couhl induce thyrocytes to express the apoptotic protein Fas, FasL and the anti-apoptotic protein Bet-2 in patients with Graves'disease. A statistically significant correlation was found between the dose of 131Ⅰ and the expression levels of Fas and FasL but not Bet-2 ,suggesting that early onset of hypothyroid after 131Ⅰ administration may be due to the increased expression of Fas and FasL in thyrocytes.
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PURPOSE: To explore the role of cell cycle regulators and apoptosis regulators in carcinogenesis of thyroid, the expression of cell cycle related proteins (cyclin D1, Ki-67) and apoptosis related proteins (survivin, caspase 3, bcl-2, p53) were investigated in follicular adenoma and follicular carcinoma of thyroid. METHODS: The following formalin-fixed paraffin embedded surgical specimens were immunohistochemically stained by avidin-biotin complex method for cyclin D1, Ki-67, survivin, caspase 3, bcl-2, p53; 15 cases of follicular adenoma (FA), 31 cases of minimally invasive follicular carcinoma (MIFC) and 12 cases of widely invasive follicular carcinoma (WIFC). RESULTS: The overexpression of six gene products in follicular neoplasms of thyroid was noted in varying frequency. Among them, increased Ki-67, caspase 3 index and overexpression of bcl-2 were noted in statistically significant, widely invasive follicular carcinoma than that of follicular adenoma and minimally invasive follicular carcinoma. CONCLUSION: These results suggest that the overexpression of Ki-67, caspase 3, bcl-2 appear to play an important role during follicular carcinogenesis of thyroid. In addition, the overexpression of these proteins is related to the differentiation of MIFC and WIFC. However, further molecular genetic studies are required to determine the interrelationships between the expression of cell cycle related proteins and apoptosis related proteins.
Subject(s)
Adenoma , Apoptosis , Caspase 3 , Cell Cycle , Cyclin D1 , Molecular Biology , Paraffin , Proteins , Thyroid GlandABSTRACT
Objective To investigate the effect of insulin signaling pathway on neuronal survival and the effect of the peptide App17 on regulating the expression of some apoptosis-related proteins in neurons of the hippocampus through intracerebrorentricular injection of streptozotocin in rats. Methods The rats were injected with App17 peptide subcutaneously three weeks after the model group was established by intracerebrorentricular injection of streptozotocin.After the four-week treament,the expressions of the apoptosis-related proteins,such as Bcl-2,Bax,CytoC in neurons of the hippocampus were tested with immunohistochemical staining and Western blotting.Results Bax,CytoC positive neurons were widely distributed in the hippocampus of the model group,and the cytoplasm was darkly stained.In contrast,the positive neurons for Bax,CytoC in hippocampus were poorly stained in the control group and the treated group,and appeared significant difference in cell counting as compared with model group.Bcl-2 positive neurons were widely distributed in the hippocampus in the control group and the treated group,and the cytoplasm was darkly stained while its positive neurons were poorly stained in the model group,and appeared significant difference in cell counting as compared with the model group.From the Western blotting clear bars could be seen in the three groups and there was a significant difference between them.Conclusions The expression of Bax,CytoC increased in the hippocampus in the rats with intracerebrorentricular injection of streptozotocin while the expression of Bcl-2 decreased.The App17 peptide could promote the rehabilitation of the abnormal expression of the three proteins to some extent.The insulin signaling pathway could affect the survival of the neurons in rats' hippocampus.