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@#Objective To investigate the effect of astragalus membranaceus（AM）injection on apoptosis and autophagy of human gastric epithelial cell line（GES⁃1）induced by enterovirus 71（EV71）. Methods GES⁃1 cells were cultured in vitro and divided into infected group（EV71 infected at a MOI of 3 and control group（no virus infected）. The morpho⁃logical changes of EV71 infected cells were observed by inverted microscope. The level of VP1 in GES⁃1 cells infected with EV71 was detected by Western blot；CCK⁃8 assay was used to detect the viability of GES⁃1 cells infected with EV71；Nuclear staining with DAPI was used to observe the morphological changes of nuclear apoptosis infected with EV71. GES⁃1 cells were divided into control group（without virus infection），infection group and AM intervention group with final concentration of 1，2. 5，5 and 10 μg／mL，respectively. Western blot was used to detect the effect of AM intervention on the expression of apoptosis⁃related proteins Caspase⁃3，PARP and autophagy⁃related proteins LC3 and P62 in GES⁃1 cells infected withEV71. CCK⁃8 method was used to detect the effect of AM intervention on the viability of GES⁃1 cells infected with EV71. Results GES⁃1 cells were round，shrunken with nuclear pyknosis and uneven size；VP1 level increased（t = 41. 56，P < 0. 01），cell viability decreased（t = 19. 07，P < 0. 01），Caspase⁃3 and PARP proteins were cut off（t = 35. 29 and 3. 648， P < 0. 01 and 0. 021 8，respectively），LC3Ⅱ／LC3Ⅰ ratio increased（t = 10. 16，P = 0. 000 5）and P62 protein was degraded（t = 68. 68，P < 0. 01）；AM inhibited the degradation of Caspase⁃3，PARP and P62 proteins induced by EV71 （t = 52. 66，59. 60 and 40. 22，respectively，each P < 0. 01）and increased the ratio of LC3Ⅱ／LC3Ⅰ（t = 5. 521，P = 0. 005 3），andreducedtheinhibitoryeffectofEV71ontheviabilityofGES⁃1cells（t =4. 420，P =0. 0115）. Conclusion EV71 infection induced apoptosis of GES⁃ 1 cells and AM intervention inhibited EV71 induced apoptosis by inhibiting EV71 induced autophagy.
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Objective To investigate the prevention effect of curcumin loaded nano - liposomes on diabetic cardiomyopathy. Methods The curcumin-loaded nano-liposomes were prepared by Film dispersion and ultrasonic hydration technology and their quality inspections were also investigated.Sixty SD rats were randomly divided into normal control group,model control group,blank and curcumin-loaded nano-liposomes group ( n＝15). Diabetes model was induced by intraperitoneal single injection of STZ(70 mg·kg-1).After two weeks of STZ injection,the rats with model control were used for this study.The curcumin loaded nano-liposomes treatment group rats were treated with curcumin loaded nano-liposomes ( 5 mg·kg-1) via caudal vein administration for 12 weeks (three times a week).Rats of normal control group,blank nano-liposomes treated group and model control group were administrated equivalent volume of 0. 9% sodium chloride solution or blank nano-liposomes solution. After treatment for 12 weeks,the experimental animals underwent ultrasonic heart function examination.Then the rats were sacrificed and their hearts were arrested after saline perfusion. The myocardial cell collagen volume fraction ( CVF) and apoptosis index were detected. Results Curcumin loaded nano-liposomes showed good morphology and curcumin encapsulation efficiency ( 88. 37 ± 1.21) %with high stability and dispersibility. From the animal experiments, the evaluation indexes in curcumin loaded nano-liposomes treated group including LVIDd and LVFS were significantly higher than model control group and nano-liposomes treated group(P<0.05),and the LVPW,CVF and apoptosis index were significantly lower than model control group and nano-liposomes treated group(P<0.05). Conclusion Curcumin loaded nano-liposomes can improve the cardiac function of diabetic rats by reducing the fibrosis and apoptosis index of myocardial cells in diabetic rats, which could be used to prevent the diabetic cardiomyopathy.
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Objective@#To investigate the effects of sIL-13Rα2 on the apoptosis of goblet cell in nasal mucosa of allergic rhinitis rats.@*Methods@#Forty healthy male Wistar rats were randomly divided into 4 groups (10 rats per group): control group (group A), AR group (group B), sIL-13Rα2 group (group C) and triamcinolone acetonide group (group D). Ovalbumin (OVA) and aluminum hydroxide were used to establish the AR rat model. After the establishment of AR rat models, 50 μl PBS, 100 μg/50 μl IL-13Rα2 and 3.5 μg/50 μl triamcinolone acetonide were respectively dropped into each nasal cavity of every rat two times a week from 4 to 10 week in group B, group C and group D. Group A was operated with saline instead of OVA. The nasal mucosa tissues were collected at 24 h after the final administration. AB-PAS staining method was used to detect the quantity and secretion of goblet cells in the nasal mucosa tissue of all groups. Immunohistochemistry method was used to detect the expression of Bax proteins.Apoptosis was detected by TUNEL method.ANOVA analysis was used to compare multiple groups, and LSD-t test was used to compare the two groups.Pearson correlation analysis was used to analyze the correlation between the Bax positive cell rate of goblet cells and the rate of apoptotic cells. The difference was statistically significant with P<0.05.@*Results@#Compared with group A, there were more goblet cells and hypersecretion of mucus in the nasal mucosa tissue of rats in group B while fewer in group C. The goblet cells in group C and group D were significantly fewer than that in group B (0.639 00±0.831 vs 0.956 7±0.980, 0.661 90±0.657 vs 0.956 7±0.980, t value was 2.748, 2.767, respectively, all P<0.05). The immunohistochemistry results showed that the positive expression rates of Bax protein in goblet cells of group C and group D were significantly higher than that in group B (0.880 2±0.125 vs 0.568 7±0.953, 0.938 4±0.200 vs 0.568 7±0.953, t value was -2.292, -2.685, respectively, all P<0.05). The apoptosis rates of goblet cell in nasal mucosa of group C and group D were also significantly higher than that in group B (0.516 0±0.079 vs 0.274 0±0.056, 0.535 4±0.829 vs 0.274 0±0.056, t value was -17.671, -2.225, respectively, all P<0.05). The expression of Bax protein and apoptosis of goblet cells were positively correlated (r=0.859, P<0.01).@*Conclusion@#sIL-13Rα2 can induce apoptosis of the goblet cells in nasal mucosa of allergic rhinitis rats, by inhibiting IL-13 and up regulating Bax.
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Objective To assess the effects of Jinghou Zengzhi Recipe(JHZZR),a Chinese prescription with the action of tonifying Qi and blood ,on the ovary apoptosis and expression of Bim in ovarian granulosa cells (GCs) of controlled ovarian hyperstimulation(COH) rats , and analyze the possible therapeutic mechanism. Methods A model of COH rats were prepaered and 30 female SD rats were randomly divided into 5 groups , including control group,positive control group,low,medium and high concentration group in six rats in each group. The apoptosis index(AI)in ovarian GCs were detected by TUNEL ,and the expression of Bim by qPCR. Results The AI of ovarian GCs in high and medium concentration group were obviously lower(P 0.05)comparing with control group. The mRNA levels of Bim were lower(P0.05)during the three concentration groups in Bim mRNA. Conclusion JHZZR can inhibit the ovarian GCs apoptosis of COH rats through decreasing the expression of Bim mRNA ,which improve the quality of ovarian follicle.
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Objective To assess the effects of Jinghou Zengzhi Recipe ,a Chinese prescription with the action of tonifying Qi and blood,on the expressions of Bcl-2,Bax and Caspase3 in ovarian granulosa cells of con-trolled ovarian hyperstimulation(COH)mice. Methods A model of COH mice was prepared and 30 female KM mice were divided into blank group,model group and treatment group. The expressions of Bcl-2,Bax and Cas-pase3 were detected by Real-time Quantitative PCR and Western Blot. Results The mRNA and protein levels of Bax and Caspase3 were lower(P<0.05),while the protein levels of Bcl-2 were significantly higher(P<0.05)in the treatment group than those in the model group. There was no significant differences between the treatment group and blank group in the mRNA and proteins. Conclusion Jinghou Zengzhi Recipe can inhibit the ovarian granulo-sa cellular apoptosis of COH mice through increasing the expression of Bcl-2 protein as well as decreasing the ex-pressions of Bax and Caspase3 mRNA and proteins to nearly the natural level,which improves the quality of ovari-an follicle.
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Objective To observe the mechanisms of bone marrow mesenchymal stem cells (BMSCs) transfected with neurotrophin-3 in treating nanoparticle repaired spinal cord ischemia-reperfusion (I/R) injury of adult rat. Methods BMSCs were transfected into NT-3 with nanoparticle carrier and transplanted into 80 adult rats on the spinal cord I/R injury model. Then the rats were divided into four groups: sham group, NS group, BMSCs and BMSCs+NT-3. The BBB scoring system, the neurons cell apoptosis, levels of malondialdehyde (MDA) and myeloperoxidase (MPO) were observed and compared between the four groups. Results The BBB score of the BMSCs+NT-3 group was significantly higher than those of BMSCs and NS group , but was lower obviously than the Sham group. TUNEL-apoptosis cells were significantly decreased in the BMSCs + NT-3 group compared to BMSCs and NS groups, but was more significantly lower than the Sham group. The contents of MDA and MPO of the NS group were significantly higher than the BMSCs group. The contents of the BMSCs + NT-3 group were lower than those of the BMSCs group, but lower significantly than the Sham group (P < 0.05). Conclusion BMSCs transfected with NT-3 with nanoparticle carrier could be induced each other. When transplanted into SCI , they can repair spinal cord by alleviating neuron cells apoptosis and inhibiting the lipid peroxidation of free radicals.
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Objective To investigate the effects of leflunomide(A771726) on the apoptosis of the rat glomerular mesangial cells and its possible mechanism .Methods The cultured rat glomerular mesangial cells were divided into the normal control group (adding DMEM culture solution containing 5% fetal calf serum) ,A771726 group(adding 50 A771726 on the basis of control group , making it a concentration as 50 μg/mL) ,LY294002 group (adding LY294002 on the basis of normal control group ,making its final concentration as 2 μg/mL) and the LY294002+A771726 group(first adding 2μg/mL LY294002 to interfere the mesangial cells for 2 h ,then adding A771726 50μg/mL) .After 48 h intervention ,its influence on mesangial cell apoptosis in each group were measured by flow cytometry .The expression of mTOR was measured by immunofluorescence .The expression of Caspase‐3 was measured by Western‐blot .Results Compared with the normal control group ,the apoptosis rate of rat glomerular mesangial cells in the A771726 group ,LY294002 group and LY294002+A771726 group were significantly increased ,the expression of mTOR was decreased ,the expression of Caspase‐3 was increased;compared with the A771726 group ,the apoptosis rates of rat glomerular mesangial cells in the LY294002 group and LY294002+ A771726 group were significantly increased ,the expression of mTOR was significantly de‐creased ,the expression of Caspase‐3 was significantly increased;compared with the LY294002 group ,the apoptosis rate of rat glo‐merular mesangial cells in the LY294002+ A771726 group was significantly increased ,the expression of mTOR was significantly decreased ,while the expression of Caspase‐3 was significantly increased .Conclusion Leflunomide may induce the rat mesangial cells apoptosis through down - regulating the PI3K/Akt/mTOR signaling pathway ,moreover ,leflunomide may synergize with LY294002 to induce rat mesangial cells apoptosis .
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Objective: To observe the effects of steroidal saponin YB16 of Ypsilandra thibetica on cell apoptosis of A549 human lung cancer cells, and explore the mechanism of apoptosis. Methods: Cells were cultured in vitro and treated with different concentration of YB16; The proliferation of A549 cells was examined by MTT method; Cells fluorescence changes of A549 were examined by acridine orange (AO) and JC-1 staining; PI, Annexin V-FITC/PI double staining, and JC-1 staining were examined by flow cytometry. Intracellular level of reactive oxygen (ROS) was determined by DCFH-DA assay, the expression of YB16 on apoptosis-related proteins was examined by Western blotting. Results: Steroidal saponin YB16 of Y. thibetica could inhibit A549 cells growth significantly with dose-effect relationship (P < 0.05); With the increasing of drug concentration, cell morphology changed significantly compared with the control group; Cell apoptosis rate increased with the different concentration of YB16 (P < 0.05); Mitochondrial membrane potential decreased significantly; ROS level of cells was increased with a significant difference (P < 0.05). Apoptotic protein, such as Bcl-2 expression of anti-apoptotic protein, was decreased, and the expression of pro-apoptotic protein Bax and cleaved caspase-3 was increased treated by YB16 after 24 h (P < 0.05). Conclusion: Steroidal saponin YB16 of Y. thibetica could inhibit the cell proliferation, reduce the mitochondrial membrane potential, enhance ROS level of A549 cells, and promote apoptosis. Therefore, it has a good anti-tumor activity.
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Objective Observe the effect of Total paeony glycoside(TPG)has on the T-cell immune action and apoptosis process of Oral lichen planus patients and analyze the mechanism.Methods 20 OLP patients are recruited into this experiment, each is treated with TPG,and their clinical effect is taken notes of.Observe the apoptosis of OLP lesions from each of the patients before and after the treatment,compare the apoptosis condition and the expression of CD4 + ,CD8 + ,and the CD4 +/CD8 + ratio,to analyze the role TPG plays on the immune expression of T-cell and the effect on apoptosis process.Results TPG improve the clini-cal manifestation of OLP patients significantly,resulting in the decrease of reticulum and erosive areas,and the pain index is de-creased obviously.The expression of CD4 + and CD8 + are decreased in OLP lesions after the treatment of TPG.Conclusion TPG may induce the apoptosis of the T-cell in lamina propria of the OLP lesions to regulate the T-cell immune action of OLP patients, and slow the development of inflammatory process of OLP,and in that way,TPG shows its potential in treating OLP,and this ex-periment can provide the primary evidence.
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Objective: To explore the effect of lavage with artiifcial cerebrospinal lfuid on neural cell apoptosis and the extracellular regulated kinase (ERK) pathway atfer traumatic brain injury. Methods: A total of 192 SD rats were randomly divided into a sham group, a traumatic brain injury model group, a local artiifcial cerebrospinal lfuid group, and a local saline group. Each group was divided into 4 sub-groups by the sacriifced time at 6 h, 12 h, 1 d and 3 d atfer the operation. hTe phosphorylation of extracellular regulated kinase 2 (P-ERK2), TNF-α and cellular apoptosis were examined. Results: hTe levels of P-ERK2 protein and TNF-α protein, as well as the number of apoptotic cellsat each time point in the local artiifcial cerebrospinal lfuid group were lower than those in the model group or in the saline group (P<0.05). Conclusion: Lavage with artificial cerebrospinal fluid can reduce apoptosis of neural cells after brain injury through the ERK pathway.
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<p><b>OBJECTIVE</b>To investigate the effect of simulated microgravity and carbon ion irradiation (CIR) on spermatogenic cell apoptosis and sperm DNA damage to the testis of male Swiss Webster mice, and assess the risk associated with space environment.</p><p><b>METHODS</b>Sperm DNA damage indicated by DNA fragmentation index (DFI) and high DNA stainability (HDS) was measured by sperm chromatin structure assay (SCSA). Apoptosis of spermatogenic cells was detected by annexin V-propidium iodide assay. Bax (the expression levels of p53) and proliferating cell nuclear antigen (PCNA) were measured by immunoblotting; p53 and PCNA were located by immunohistology.</p><p><b>RESULTS</b>HDS, DFI, apoptosis index, and the expression levels of p53 and Bax were detected to be significantly higher in the experimental groups (P<0.05) compared with those in the control group; however, the PCNA expression varied to a certain degree. p53- and PCNA- positive expression were detected in each group, mainly in relation to the spermatogonic cells and spermatocytes.</p><p><b>CONCLUSION</b>The findings of the present study demonstrated that simulated microgravity and CIR can induce spermatogenic cell apoptosis and sperm DNA damage. Sperm DNA damage may be one of the underlying mechanisms behind male fertility decline under space environment. These findings may provide a scientific basis for protecting astronauts and space traveler's health and safety.</p>
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Animals , Male , Mice , Apoptosis , Radiation Effects , Carbon , Cell Proliferation , Radiation Effects , DNA Damage , Heavy Ions , Immunohistochemistry , Random Allocation , Sperm Count , Spermatogenesis , Radiation Effects , Spermatozoa , Radiation Effects , Testis , Radiation Effects , Weightlessness SimulationABSTRACT
Objective To study the effects of emodin on apoptosis and mRNA and protein of apoptosis inducing factor (AIF) and Endonuclease G (Endo G) in human bladder cancer cells BIU87,and to investigate the anticancer mechanism of emodin. Methods The BIU87 cells were divided into 4 groups,control group,Z-VAD-FMK group,emodin group,emodin combined with Z-VAD-FMK group.The effects of different concentrations of emodin at different action time on cells proliferation of BIU87 in vitro culture were measured by methylthiazole (MTT) chromatometry,the cells apoptosis were detected by flow cytometry,and expression of AIF and Endo G were examined by reverse transcription PCR (RT-PCR) and Western blot.Results MTT assay demonstrated that the higher concentration of emodin and the longer action time,the more significant inhibition of tumor cell growth.Based on the IC50 value,80 μmol/L and 72 h of emodin intervention were selected as an intervention condition.The apoptosis rate in emodin group (44.57 ± 1.52 ) ％was significantly higher than that in emodin combined with Z-VAD-FMK group (35.58 ± 1.61 ) ％ ( P ＜0.01).RT-PCR and Western blot showed that the mRNA and protein of AIF in emodin combined with Z-VAD-FMK group,emodin group,control group,Z-VAD-FMK group were ( 1.74 ± 0.11 ) and (2.59 ±0.13),(1.36±0.08) and (1.89±0.14),(0.37 ±0.02) and (0.53±0.11),(0.42 ±0.06) and (0.44 ± 0.07),respectively.There were significant differences between emodin group and the other groups (P ＜0.01 ).The mRNA and protein of Endo G in emodin combined with Z-VAD-FMK group,emodin group,control group,Z-VAD-FMK group were (2.28±0.15) and (3.31 ±0.36),(1.85 ±0.13) and (2.15 ±0.27),(0.53 ±0.07) and (0.71 ±0.16),(0.61 ±0.04) and (0.67 ±0.22),respectively.The differences were significant between emodin group and the other groups ( P ＜ 0.01 ). Coneltusion Emodin can upgrade the expression of AIF and Endo G in bladder cancer cells BIU87,which can induce apoptosis through Caspase-independent pathway.
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(Ver)-induced human retinal pigment epithelium (RPE) cells apoptosis.hours to induce RPE cells apoptosis.The expression of apoptotic effector gene caspase-3 was assessed by reverse transcription polymerase chain reaction (RT-PCR).Single cell was measured using fluorescence indicator Fura-3/AM with MetaFluo4.5/coolsnapfx/IX70 intracellular Ca2+ fluorescence imaging system.RPE cells and it significantly increased after co-cultured with Ver.The fluorescence in resting RPE cells was strong and distributed throughout the cells.The nucleus appeared more fluorescent than the cytoplasm.Calcium fluorescence of RPE cells attenuated after co-cultured with Ver.[Ca2+]i homeostasis might play pivotal roles in Ver-induced RPE cells apoptosis.
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Objective:To study the biological effects of grape seed extract(GSE) on the human ovary cancer cells and explore its molecular mechanism. Methods: Human ovary cancer cells line SKOV3 were cultured.They were put into 96-well plate and added with solutions of GSE of the terminal concentrations of 25,50,100,200,400 ?g/ml respectively,and 24,48,72 hours later cell growth curve was drawn. MTT assay was used for other SKOV3 cells co-incubated with GSE of the terminal concentration of 25,50,100,200,400 ?g/ml respectively so as to calculate the proliferation inhibition rate. SKOV3 cells were treated with GSE of the concentration of 25 ?g/ml for 24,48 hours respectively. Flowcytometry(FCM)was used to analyze the DNA cycle and TUNEL was used to calculate the apoptotic rate. Annexin-V labeling method was used to detect the positive rate of apoptotic cells. SKOV3 cells were treated with GSE of the concentration of 25 ?g/ml for 24,48,72 hours.Western blotting was used to detect the protein expression of caspase-3. Results:GSE dose-dependently inhibited the proliferation of the SKOV3 cells in a dose-dependent manner.Treated by GSE,the progress of cells at stage into G2/M stage was inhibited.Tunel showed that treated by GSE (25 ?g/ml)for 24、48 h the apoptotic rates of the cells were 31.98%and 45.78%, respectively. Annexin-V showed that after incubation with GSE (25 ?g/ml)for 12,24,48 h,the apoptotic rate were 6.71%,19.05%, 36.55%,respectively. Western blotting showed that the caspase-3 protein expression were up-regulated.Conlusion:GSE inhibits the proliferation of malignant human ovary cancer cells and induces their apoptosis. Expression of caspase-3 protein may be related to cell growth inhibition and the apoptosis mediated by GSE in vitro.
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Objective To explore the treatment effect of Zuogui pill (ZGP) on immune premature ovarian failure rats.Methods Female mice model of POF was established by multiple sites subcutaneous injection of zona pellucida 3,and treated with different dosage ZGP (low, middle and high),with prednisone and diaethylstilbestrol as positive control. We detecteed apoptosis rate of granulosa and oocyte of murine ovaries by tunel method and observd the change of sexual cycle and weight. Results Compared with the control group,the weight of model group rat in the third to eighth weeks was significant difference (P
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The known members of inhibitor of growth (ING) gene family are considered as candidate tumor suppressor genes. ING4, a novel member of ING family, is recently reported to regulate brain tumour angiogenesis through transcriptional repression of NF-?B-responsive genes, induce G2/M arrest by the increased p21 expression in a p53-dependent manner, suppress the loss of contact inhibition and represses activation of the hypoxia inducible factor, which plays an important role in the progression of tumorigenesis. However, seldom studies about ING4 inducing tumor cells apoptosis were reported.The C6 cells (mouse glioma cells) were infected respectively with the blank adenovirus carrying GFP (Ad) and the recombinated Ad-hING4-His, then RT-PCR assay was used to detect the transcriptions of hING4, as well Western-blotting assay was ued to detect the expressions of hING4. The effects of hING4 expression upon C6 cells were observed, and the growth curve was drawed and tumor control rates were calculated. The C6 cells, which were affected by blank Ad and Ad-hING4-His, were respectively observed by LSCM (laser scan confocal microscope) and transmission electron microscope (TEM), detected by flow cytometry; and the genomic DNA of both groups were extracted and electrophoresised in agarose gel to examinate the DNA fragments. The results showed hING4 can significantly inhibit the growth of C6 cells by promoting the cell’s apoptosis, which probably is the first one to prove this property of ING4.The experimental and theoretical foundation for gene therapy for gliomas with ING4 in the future was established.
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AIM: To investigate protect effect of the NO donating-oleanlic acid derivatives on hepatic fibrosis in rats. METHODS: The rat model with early hepatic fibrosis was induced by carbon tetrachloride (CCl_4). ZCⅡ_2 at the dose of 128 and 64 mg?kg -1 had been given orally for 30 days. Serum level of the total protein (TP), albumin (ALB), the albumin/globulin (A/G) and ALT, AST. The hyaluronic acid (HA), laminin (LN), the procollagen type Ⅲ (PCⅢ) and the level of MDA, GSH-Px in liver tissues were determined. Pathological examination to reveal the extent of liver damages was observed. [WTHZ]RESULTS: ZCⅡ_2 at the dose of 128 mg?kg -1 increased the serum TP, ALB, and A/G and decreased HA, LN, PCⅢ, ALT, and AST more significantly than the model group, and hepatic pathological injury was abated to some degree. CONCLUSIONS: ZCⅡ_2 at the dose of 128 mg?kg -1 can attenuate liver damages, protect against lipoperoxidation and attenuate hepatic fibrosis induced by CCl_4.
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BACKGROUND: Inducible nitric oxide synthase (iNOS) has been detected in a number of pathologic conditions in the central nervous system. This study was investigated the patterns of iNOS expression in the neuronal PC12 cell and the effects of nitric oxide on the apoptosis of PC12 cells. METHODS: The stimulating agents for induction of iNOS expression in PC12 cells were bacterial lipopolysaccharide (LPS), tumor necrosis factor-alpha (TNF-), and interferon-gamma (IFN-). RESULTS: The expression iNOS mRNA and protein in PC12 cells stimulated with LPS/TNF-/IFN- were profoundly increased. The expression of iNOS mRNA arose at 6 hours, peaked at 12 hours, and declined to 48 hours after LPS/TNF-/ IFN- treatment. iNOS protein was increased up to 24 hours in LPS/TNF-/IFN- treated PC12 cells while the expression of nNOS was unaffected. Accumulation of NO derivatives in the culture media was markedly increased at least at up to 48 hours after LPS/TNF-/IFN- treatment. The induction of iNOS expression and NO production in differentiated PC12 cells was correlated with apoptotic cell death judged by transmission electron microscopy and DNA fragmentation from the results of the Terminal deoxynucleotidyl-transferase-mediated dUDP biotin nick end-labeling (TUNEL) method. After treatment with NOS inhibitor, N-monomethylarginine (NMMA), a profound decrease in NO production by LPS/TNF-/IFN- treated PC12 cells was noted. And the LPS/TNF-/IFN- induced apoptosis was prevented by the NMMA treatment. CONCLUSIONS: From the above results it is concluded that the expression of iNOS in differentiated PC12 cells is induced by the combined application of LPS, TNF-, and IFN-. And the apoptosis of cultured PC12 cells is mediated by iNOS-derived NO.
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Animals , Apoptosis , Biotin , Cell Death , Central Nervous System , Culture Media , DNA Fragmentation , Interferon-gamma , Microscopy, Electron, Transmission , Necrosis , Neurons , Nitric Oxide Synthase Type II , Nitric Oxide , PC12 Cells , RNA, Messenger , Tumor Necrosis Factor-alphaABSTRACT
Objective:To observe anti-DR5 monoclonal antibody on apoptosis and CDC effect in EC109 cells.Methods:The anti-DR5 monoclonal antibody was prepared by hybridoma technique.Tumoricidal effects and complement-dependent cytotoxicity of the McAb to EC109 cells was screened by MTT assay.The apoptosis of EC109 cells was detected by flow cytometry with annexin Ⅴ-FITC/PI staining.Morphological change of EC109 was observed by microphotograph.Results:An anti-DR5 monoclonal antibody was obtained.It induced apoptosis of EC109 dose dependently.The cytotoxic action was notably enhanced by addition of complement.The cells growth inhibition ratios reached 83.04%.The apoptotic body and cathepsis were seen in microphotograph.Conclusion:The anti-DR5 monoclonal antibody could induce EC109 cells apoptosis and cause the complement dependent cytotoxic (CDC) effects powerfully.
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Objective:To experimental study the inhibiting effects of SBHL on the growth of cervical cancer cells and to investigate the mechanisms.Methods:The proliferation of ME180 cells was observed by MTT assasy; The changes of cells cycles and apoptosis of ME180 cells treated with SBHL were analyzed by agar gel electrophoresis and FCM assay;. The changes of the ultra structure of cells was observed by Transmission eleconmicrograph.Results:The proliferation of ME180 cells was inhibited after treating on SBHL for 24 h.The inhibiting effect increased granually following the concentration of SBHL(P