共查询到20条相似文献，搜索用时 31 毫秒
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AIM: To study the inducing effect of human mutant p27 gene on apoptosis of the colorectal cancer cells. METHODS: The recombinant adenovirus Ad-p27mt was constructed to infect the colorectal cancer cell SW480. The inducing effect of Ad-p27mt on apoptosis in colorectal cancer cells was measured by flow cytometry, DNA fragment analysis and TUNEL method. RESULTS: Ad-p27mt was successfully constructed. When the multiplicity of infection (MOI) was ≥50, the infection efficiency reached 100%. After 24 h of infection, there was an apoptotic hypodiploid peak observed by flow cytometry before G1 and there were apoptotic characteristic bands in the DNA electrophoresis. The apoptotic index detected by TUNEL method was 82.6±3.2 (Ad-p27mt group) and 5.0±3.5 (control group), respectively, the difference of which was significant (P<0.01). CONCLUSION: Human mutant p27 gene transfection effectively induces apoptosis in the colorectal cancer cells. 相似文献
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JIANG Ying-juan ZENG Yao-ying WANG Tong ZHAO Jing-xian XING Fei-yue WANG Xi-chao LIANG Pei-yan 《园艺学报》2006,22(5):846-850
AIM: To study the changes of mitochondrial membrane potential (△ψm) and mitochondrial mass in apoptosis of Jurkat cells induced by camptothecin (CPT). METHODS: Jurkat cells were treated with CPT. Annexin V-FITC/propidium iodine (PI) double stainig was used to detected early stage of apoptosis and PI staining for analyzing the cell cycle. Jurkat cells were stained by annexin V-PE/DiOC6(3) to detect changes of △ψm. The mitochondrial mass was measured by cytometry with NAO staining. RESULTS: 6 h after treated with 10 μmol/L CPT, the rate of early apoptotic cells (22.59±1.04)% had significantly difference compared with control group (3.93±0.73)% (P<0.01). The necrotic rate (2.48±0.53)% had no significant difference to that in control group (2.78±0.63)% (P>0.05). Apoptotic peak appeared obviously after treated with CPT, the percentage of late apoptotic cells (13.58±0.97)% had distinctly difference compared with control group (3.18±0.51)% (P<0.01). The cells in G0/G1 phase (48.14±0.96)% were much higher than that in control group (44.09±0.43)% (P<0.01). Mitochondrial depolarization was very obviously in CPT group. The percentage of annexin V+DiOC6 (3)- cells was (19.47±0.69)%, while in control group, was (4.21±0.40)% (P<0.01). Mitochondrial mass in CPT group was significantly lower than that in control group, the percentage of NAO+ cells (74.77±1.66) % had significantly difference compared with control group (92.24±1.41)% (P<0.01). CONCLUSION: During the process of CPT-induced apoptosis in Jurkat cells, mitochondrial depolarization was very obviously and mitochondrial mass decreased, indicating that the process of apoptosis is nearly related to the mitochondrial pathway. 相似文献
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LIU Hong-mei ZHEN Guo-hua ZHANG Zhen-xiang CAO Yong YANG Shi-fang XU Yong-jian 《园艺学报》2008,24(5):931-935
AIM: To observe the apoptosis of alveolar wall cells in the papin-induced rat emphysema, and to explore the role of apoptosis of alveolar wall cells in the pathogenesis of emphysema. METHODS: Rats were randomly divided into three groups: normal control group, papain only group, papain+irradiation group. Morphology of lung tissues was assessed. TUNEL assay was used to determine the apoptotic cells. Immunohistochemistry was performed to determine the expression of PCNA, Bax and SP-C in the lung alveolar wall cells. SP-C immunofluorescence stainging was performed to identify the type II alveolar cells in the TUNEL positive cells. RESULTS: Destruction of alveolar wall and loss of the alveolar unit were observed in papain only group and papain+irradiation group. There was significant difference between rats of papain+irradiation group and papain only growp in the mean linear interval (MLI), the number of alveolar counted per unit area (MAN) and mean alveoli area (MAA), respectively (P<0.01). The proliferation index(PI), apoptosis index (AI) and the percentage of Bax in the papain only group and papain+irradiation group were significantly greater than that in the normal control group (P<0.01). However, the percentage of SP-C positive cells was significantly lower in the papain only group and papain+irradiation group compared with the normal control (P<0.01). Moreover, the PI, AI and the percentage of Bax in the papain+irradiation group was higher than those in the papain only group. The percentage of SP-C positive cells in the papain+irradiation group was lower than that in the papain only group. Most of the TUNEL-positive cells expressed the SP-C. CONCLUSION: Apoptosis of rat alveolar wall cells, especially type-II cell may be involved in the pathogenesis of emphysema. Upregulation of Bax expression may lead to alveolar wall cells apoptosis in papain-induced emphysema. 相似文献
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AIM: To investigate the anti-tumor efficacy of angiogenic inhibitor three hrombospondin-1 type repeats (3TSR) on gastric cancer. METHODS: Human gastric carcinoma cell line SGC-7901 was inoculated subcutaneously to BALB/c mice, and then the mice were divided into two groups (8 mice each): control group and 3TSR group. After administration of 3TSR by intraperitoneal injection for 3 weeks, mice were sacrificed. The tumor volume and percentage of necrotic area were detected. The micro-vessel index and cell proliferation index were detected by immunohistochemistry method. The apoptosis rate of gastric carcinoma cells was measured by TUNEL method. The vascular endothelial cell apoptosis rates were detected by CD31/TUNEL/DAPI staining. RESULTS: The tumor volume in 3TSR group was (648.34±126.91)mm3, significantly lower than that in control group (P<0.05). The percentage of tumor necrosis area in 3TSR group was (39.6±7.8)%, almost increased by 69.2% than that in the control. Average micro-vessel numbers and micro-vessel area in 3TSR were 12.8±4.1 and (689.3±118.6) μm2, respectively, significantly lower than those in the control. The proliferation index and apoptosis rate in 3TSR group were (40.0±7.1)% and (3.4±1.2)%, respectively. No difference between 3TSR group and the control was observed. The endothelial cell apoptosis rate in 3TSR group was (11.6±2.8)%, significantly higher than that in control group (2.9±1.5)%. CONCLUSION: 3TSR inhibits tumor angiogenesis, remarkably reduces tumor volume, average micro-vessel and increased tumor necrosis. 3TSR shows no direct inhibitory effects on gastric cancer cells. The antiangiogenesis effects of 3TSR on gastric carcinoma may be due to the induction of apoptosis of vascular endothelia cells. 相似文献
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AIM: To investigate the balance of Treg/Th17 in synovium of collagen-induced arthritis (CIA) and the impact of tumor necrosis factor α(TNF-α) blockage therapy. METHODS: Rat CIA model was established by bovine II collagen injection. The pathological score was evaluated by HE staining and toluidine blue staining. The TNF-α level in plasma was measured by ELISA. The expression of Treg/Th17 in synovium was detected by double staining immunofluorescence. RESULTS: The plasma level of TNF-α in CIA group was significantly higher than that in control group and TNFR-Fc treatment group (P<0.01), whereas no significant difference was found between TNFR-Fc treatment group and control group (P>0.05). No significant difference between CIA group and control group in the ratio of CD4+Foxp3+Treg cells/CD4+ cells in synovium (23.12%±4.93% vs 24.66%±5.82%, P>0.05) was observed, whereas the ratio in TNFR-Fc treatment group was significantly increased(33.07%±5.14%). The ratio of CD4+RORγt+Th17 cells/CD4+ cells in CIA group was significantly higher than that in control group and TNFR-Fc treatment group (9.74%±2.23% vs 1.00%±0.59%, 5.63%±1.76%, P<0.01). CONCLUSION: Differentiation disturbance of Treg/Th17 exists in the synovium of CIA rats. TNFR-Fc may restore the balance of Treg/Th17 by inhibiting Th17 cell differentiation and inducing the production or accumulation of Treg. 相似文献
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AIM: To detect the changes of autophagy in adipose cells under starvation, and to clarify the effects of autophagy on the cell survival and apoptosis under starvation. METHODS: Rapamycin (RAP) was applied to promote autophagy of adipose cells. These cells were then incubated under oxygen-glucose deprivation (OGD) condition. After exposure of the cells to OGD, the changes of autophagy and apoptosis were determined by Western blotting, transmission electron microscopy and TUNEL assay. RESULTS: Compared with the control cells, OGD-challenged cells had much higher level of autophagy. The apoptotic rate in OGD group was much higher than that in control group, which was reflected by increased protein level of activated caspase-3 and percentages of TUNEL positive cells. Preconditioning with RAP effectively improved OGD-induced autophagy, but did not affect the cell survival and apoptosis under normal condition, and obviously decreased the apoptotic rate of the cells under OGD condition. CONCLUSION: Autophagy protects adipose cells against starvation-induced apoptosis. Promotion of autophagy is helpful for attenuating starvation-induced apoptosis of the cells under OGD condition. 相似文献
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AIM: To examine whether ischemic preconditioning (IPC) can protect against apoptosis in CA1 subfield of hippocampus following reperfusion of a lethal ischemia in rats and explore the role of IPC by inhibiting the expression of p53 in this process. METHODS: Wistar rats were used in the experiment. A global ischemia/reperfusion model was induced by 4-vessel occlusion. The rats were divided into the following three groups randomly: (1) ischemic preconditioning group (IPC group); (2) ischemia/reperfusion group (IR group); (3) control group. The histopathological changes, the percentage of apoptosis and the expression of p53 gene in CA1 region of rat hippocampus were examined by HE staining, FCM, RT-PCR and immunohistochemistry techniques. RESULTS: The neuronal density of CA1 region in IPC group ［(217±9)/0.72 mm2］ was significantly higher than that in IR group ［(29±5)/0.72 mm2, P<0.01］. The percentage of apoptotic neurons in IPC group (2.07%±0.21%) was lower than that in IR group (4.26%±0.08%), P<0.01. Compared with IR group, the expression of p53 gene in IPC group was significantly weakened. CONCLUSION: Ischemic preconditioning protects the ischemic neurons in CA1 region of rat hippocampus by inhibiting the expression of p53 gene. 相似文献
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AIM: To observe the effect of leptin (LEP) on hypoxia-reoxygenation induced apoptosis in L02 cells.METHODS: In the experiment, L02 cell injury was induced by hypoxic air (95%N2 and 5% CO2). The cultured L02 cells were divided into hypoxic 12 h group (IR group) alone, normal control group and the hypoxic plus leptin (100 μg/L, 200μg/L, 400 μg/L, 800 μg/L and 1 600 μg/L) treatment groups in vitro. Flow cytometry, terminal dUTP nick-end labeling (TUNEL) assay and fluorescent quantitative PCR were used to measure the changes of apoptosis in L02 cells and expression of Fas/FasL mRNA.RESULTS: (1) The percentage of L02 cells apoptosis and positive TUNEL cells significantly increased in IR group compared to control group (P<0.01). When L02 cells were treated with LEP, the percentage of cell apoptosis and positive TUNEL cells were decreased compared to IR group. (2) Compared to control group, the Fas/FasL mRNA expression significantly increased in IR group (P<0.01). When L02 cells were treated with LEP, the Fas/FasL mRNA expression decreased compared to IR group, the effect of LEP at concentration of 400 μg/L was obviously (P<0.05). CONCLUSION: LEP decreases the apoptosis of hypoxic-reoxygenation L02 cells by down-regulation of Fas/FasL mRNA expression in L02 cells. 相似文献
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AIM: To investigate the promoting effects of survivin-siRNA on apoptosis of DU145 cells. METHODS: The DNA template coding siRNA against survivin was synthesized and recombinant plasmid pSi-sur was constructed. The recombinant and the two controls, liposome and pSi-scrambled plasmid, were transfected into DU145 cells. The expression of survivin in mRNA and protein was detected by RT-PCR and Western blotting, respectively. Apoptosis was measured by acridine orange staining, Annexin V-FITC labeled flow cytometry and TUNEL assay.RESULTS: Compared to the liposome control, the levels of survivin mRNA and protein in siRNA group were 0.28±0.07 and 0.34±0.05 (n=3, P<0.05) respectively 72 h after transfection. The apoptotic rate of the cells in pSi-sur group was significantly higher than that in two control groups as showed in acridine orange staining, Annexin V-FITC labeled flow cytometry and TUNEL assay.CONCLUSION: Survivin siRNA significantly inhibits the expression of survivin both in mRNA and protein levels, and induces apoptosis of DU145 cells in vitro. 相似文献
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WANG Hui-ying ZENG Yao-ying WANG Tong XING Fei-yue ZHAO Jing-xian JI Yu-hua 《园艺学报》2006,33(5):1020-1023
AIM: To study the effects of ultraviolet (UV) on mitochondrial functions and apoptosis in HaCaT cells. METHODS: After irradiation by UV at low dose (UVA 2 J/cm2，UVB 10 mJ/cm2) and high dose (UVA 6 J/cm2，UVB 30 mJ/cm2), HaCaT cells were cultured for 15 hours. Flow cytometry was used to measure mitochondrial membrane potential, mitochondrial mass and apoptotic rate. Annexin V-FITC/PI staining of apoptotic cells was analyzed by laser confocal microscopy. RESULTS: After UV irradiation, cell proportion with low mitochondrial membrane potential increased with irradiation doses. The proportion of control group, low dose group and high dose group were 7.94%±1.02%, 25.87%±4.55% and 39.27%±5.32%, respectively. Cells proportion with low mitochondrial mass increased with irradiation doses. The proportion of control group, low dose group and high dose group were 15.19%±1.58%, 40.36%±4.41% and 68.79%±5.46%, respectively. The hypodiploid peaks of DNA content analysis represented the apoptotic rate of HaCaT cells. The apoptotic rate of control group, low dose group and high dose group were 1.82%±0.51%, 30.16%±5.47% and 58.49%±5.98%, respectively. To analyze the cells apoptosis by staining with annexin V-FITC and PI, the results were consistent with those of DNA content analysis. Cells in control group showed almost no positive staining cells. Single annexin V-FITC positive cells in low dose group and double positive cells in high dose group were predominant, respectively. CONCLUSION: UV irradiation induces HaCaT cell mitochondrial depolarization, as well as mitochondrial mass loss. These changes are related to cell apoptosis. 相似文献
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AIM: To investigate the effect of N-acetylcystein (NAC) on oxidant stress, neuron apoptosis in the hippocampal CA1 region of rats exposured to chronic intermittent hypoxia (CIH). METHODS: 30 healthy male Wistar rats were randomly divided into three groups of 10 each, a CIH group, a NAC therapeutic group and a control group. The levels of MDA and SOD were detected by colorimetric method. Immunohistochemistry was used to examine the expression of p-JNK and TUNEL was used to detect the neuron apoptosis in the hippocampal CA1 region. RESULTS: The level of MDA in NAC group were lower than that in CIH group［(1.71±0.43) μmol/g protein vs （1.37±0.26) μmol/g protein, P<0.05)］. The activity of SOD in NAC group was higher than that in CIH group［(44.94±14.01) 103 NU/g protein vs(57.66±14.07) 103 NU/g protein, P<0.05)］. The expression of p-JNK protein and the apoptotic indices ［(0.39±0.16), (0.20±0.11)］ in NAC group were significantly lower than those in CIH group ［(0.53±0.10), (0.32±0.18), all P<0.05］. CONCLUSION: NAC protects hippocampal neuron from apoptosis by suppressing the oxidant stress in the hippocampal CA1 region and inhibiting the activation of JNK signaling pathway. 相似文献
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AIM:To study the effect of activated protein C (APC) at different concentrations on apoptosis of human umbilical vein endothelial cells (HUVECs) induced by lipopolysaccharide (LPS).METHODS:The HUVECs were induced by LPS (1.0 mg/L) as apoptotic model that was administered by different concentration of APC (10 μg/L or 50 μg/L). Meanwhile, the control group and induced apoptosis group induced by LPS (1.0 mg/L) stimulation were also set up. The changes of cellular ultrastructures were observed under electron microscope. The DNA ladder and TUNEL fluorescent staining were measured in cells. Annexin-Ⅴ/PI double staining was used to measure the cell apoptosis rate by flow cytometry. Cell survival rate was measured by MTT assay. The proliferating cell nuclear antigen (PCNA) expression levels in cells were also measured by Western blotting to reflect the proliferation of the cells.RESULTS:There were significant apoptotic changes in the cells induced by LPS, but the apoptotic changes were reduced and apoptosis rates were decreased in the cells treated with APC. Meanwhile, cell survival rate and the protein levels of PCNA were increased after APC treatment, particularly at the concentration of 50 μg/L, which showed difference when compared with those induced apoptosis group by LPS (P<0.05).CONCLUSION:APC can inhibit HUVECs apoptosis induced by LPS and promote cell proliferation, thus protect the cells from injury. 相似文献
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AIM: To study the role of hypoxia preconditioning (HP) in hypoxia-reoxygenation (HR)-induced apoptosis in neonatal rat cardiomyocytes and the possible mechanisms. METHODS: Cultured neonatal rat cardiomyocytes were divided into three groups: normal group, HP+H/R group and H/R group. Acridine orange (AO) staining was performed to detect morphological changes of apoptotic cells. Apoptosis rates of cardiomyocytes were detected by flow cytometry. Colorimetric assay was used to detect caspase-3 activity. Expression of Bcl-2 protein was detected by immunohistochemistry combined with computer image analysis. RESULTS: Apoptotic cells were detected by AO staining after hypoxia of 6 h followed by 3 h-reoxygenation. The hypodiploid apoptotic peak was detected by flow cytometry with the apoptotic rates of (29.7±5.4)%. A significantly reduced apoptotic rates of (7.8±1.3)% was detected in HP group（P＜0.01）. The caspase-3 relative activity of cardiomyocytes induced by H/R was 5.9±0.8, significantly higher than that of control group. HP markedly reduced caspase-3 relative activity to 2.6±0.5 in contrast with H/R group （P＜0.01）. Bcl-2 protein was positive in normal cardiomyocytes with an A value of 119.4±7.1. The A value of H/R group was 99.6±5.0, significantly lower than that in normal group （P＜0.01）. The A value of HP+H/R group was 126.5±6.2, significantly higher than that in H/R group（P＜0.01）. CONCLUSION: HP inhibits H/R-induced apoptosis of cardiomyocytes by improving the expression of Bcl-2 and reducing caspase-3 activity. 相似文献
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LI Min HUANG Ting-ting JIANG Shuang-yan YANG yang CHEN Sha-sha ZHANG Wen-jia ZHAO Li-juan 《园艺学报》2019,35(11):2050-2054
AIM: To investigate whether curcumin reduces hepatocyte apoptosis in the rats with non-alcoholic steatohepatitis (NASH) by inhibiting endoplasmic reticulum stress (ERS) and thus exerting a protective effect on the liver. METHODS: Male SD rats (n=30) were randomly divided into normal control group (n=10), model group (n=10) and curcumin group (n=10). NASH model was established by feeding the rats with high-fat diet for 4 weeks. The rats in curcumin group was given curcumin (200 mg/kg) daily by gavage, while the rats in model group and normal control group were given the same volume of saline. Four weeks later, the rats were killed, and their blood and liver tissues were collected. The serum levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were detected, liver histopathological changes were observed by HE staining, the expression of glucose-regulated protein 78 (GRP78) and C/EBP homologous protein (CHOP) was determined by Western blot, and apoptosis was detected by TUNEL method. RESULTS: Compared with normal control group, the levels of serum ALT and AST in model group were significantly increased, and the levels of serum ALT and AST in curcumin group were significantly lower than those in model group (P<0.05). At the same time, the steatosis and inflammation of hepatocytes in curcumin group were less than those in model group, and no obvious necrosis was observed. Compared with normal control group, the protein expression levels of GRP78 and CHOP in model group were increased, while the protein expression levels of GRP78 and CHOP in curcumin group were decreased compared with model group (P<0.01). TUNEL results showed that apoptotic hepatocytes in model group were significantly more than those in normal control group, while those in curcumin group were significantly fewer than those in model group. CONCLUSION: Hyperlipidemia induces excessive ERS in the hepatocytes, thus triggering apoptosis and leading to NASH. The mechanism of curcumin reducing hepatocyte apoptosis may be related to its inhibition of ERS. 相似文献
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AIM:To observe the effects of Helicobacter pylori(Hp) on the apoptosis of human gingival tissue.METHODS:Gingival tissue samples were taken from 30 patients without chronic periodontitis,and Hp was detected by conventional PCR.The apoptosis of the gingivival cells was detected by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) to analyze the correlation between Hp infection and apoptosis of the gingival tissues.RESULTS:The Hp positive detections were 12 in the 30 patients without periodontitis,so the positive rate of Hp in the gingival tissue samples was 40%.The gingival tissue showed a large number of apoptotic cells in Hp positive group,and less apoptotic cells in Hp negative group.The apoptotic index in Hp positive group (0.498±0.092) was significantly higher than that in normal group (0.207±0.053)(P<0.05).CONCLUSION:Hp might play a role in the apoptosis of gingival tissues. 相似文献