Effects of polyphyllin Ⅵ on the proliferation and apoptosis of glioma cells and potential mechanism / 中国药房

OBJECTIVE To investigate the effects of polyphyllin Ⅵ（PPⅥ） on the proliferation and apoptosis of glioma cells and potential mechanism. METHODS Using human glioma LN229 cells as objects， MTT assay was used to detect the survival rate after treated with different concentrations of PPⅥ [0 （control group）， 1， 2， 4， 8， 16， 32， 64 μmol/L] for different time （24， 48， 72 h）. The clone formation experiments were adopted to detect the number of cell clones and clone formation rate after being treated with different concentrations of PPⅥ [0 （control group）， 2， 4， 8 μmol/L] for 14 days. The flow cytometry and Western blot assay were used to detect the apoptotic rate of cells， the expressions of apoptosis-related protein [B cell lymphoma-2 （Bcl-2）， Bcl-2 associated X protein （Bax）， cleaved caspase-3]， and the expressions of related proteins of Fas/Fas ligand （FasL） death receptor pathway and protein kinase B （Akt）/glycogen synthesis kinase-3β （GSK-3β） pathway after being treated with different concentrations of PPⅥ [0（control group）， 4， 8 μmol/L] for 24 h. RESULTS Compared with the control group， the survival rate of cells， the number of clones and clone formation rate， the protein expression of Bcl-2， and the phosphorylation levels of Akt and GSK-3β protein were decreased significantly in different concentration groups of PPⅥ （P＜0.05 or P＜0.01）. The apoptotic rate， the protein expressions of Bax， cleaved caspase-3， Fas， FasL and cleaved caspase-8 were increased significantly （P＜0.05 or P＜ 0.01）. CONCLUSIONS PPⅥ can inhibit the proliferation and induce the apoptosis of human glioma LN229 cells， which may be related to the activation of the Fas/FasL death receptor pathway and the inhibition of the Akt/GSK-3β pathway.

Transplantation of human stem cells from the apical papilla for treating dextran sulfate sodium-induced experimental colitis / 中国组织工程研究

BACKGROUND: Stem cells from the apical papilla (SCAP) play important roles in the formation and development of dental roots. However, the immune-modulating capacity of SCAP has not been fully elucidated. OBJECTIVE: To test the therapeutic effects of transplantation of SCAP on dextran sulfate sodium-induced experimental colitis. METHODS: Twenty-four C57/BL6 mice were equally divided into four groups (normal control, positive control, SCAP treatment group, and FasL-knockdown SCAP group), and latter three groups of mice were induced to acute experimental colitis by 3% dextran sulfate sodium in drinking water. At day 3 after modeling, model mice were treated with PBS, human SCAP (2×106 cells), and FasL-knockdown SCAP via intraperitoneal injection, respectively. Inflammation was evaluated by measuring body mass and length of the colon, detecting levels of interleukin 1β, interleukin 6 and tumor necrosis factor α, as well as histological analyses at day 10 after modeling. Levels of Tregs in mesenteric lymph nodes in mice were detected using flow cytometric analysis. RESULTS AND CONCLUSION: SCAP transplantation could ameliorate the inflammation in dextran sulfate sodium-induced colitis mice, and body mass loss and symptoms were significantly improved. Pathological score and the levels of three inflammatory cytokines in the colon tissue decreased significantly. Flow cytometric analysis revealed an increased level of Tregs in mesenteric lymph nodes. Knocking down of FasL gene in SCAP abrogated the therapeutic effects of SCAP in ameliorating dextran sulphate sodium-induced colitis. Therefore, Fas-FasL pathway played an important role in the underlying mechanism of the immune-modulating capacity of SCAP.

Experimental study on the effect of hedysarum polybotys saccharides and selenizated hedysarum polybotys saccharides on oral squamous cancer cells in vitro / 口腔疾病防治

Objective@#To study the effects of hedysarum polybotys saccharides (HPS) and selenizated hedysarum polybotys saccharides (SE-HPS) on the oral squamous cancer cell line SCC25.@*Methods@#Different concentrations (0, 10, 25, 50, 100, 200, 400 μg/ml) of HPS and SE-HPS were added to SCC25 cells in the logarithmic growth stage. Cell proliferation was detected by the CCK-8 method, apoptosis was detected by flow cytometry, and apoptosis-related indexes were observed by RT-qPCR and Western blotting.@*Results @#The concentrations of HPS and SE-HPS inhibited the proliferation of SCC25 cells. The inhibitory effect of 50 μg/mL HPS and SE-HPS on the proliferation of SCC25 cells was the strongest and was time-dependent. The inhibition effect significantly increased within 48 h, and the effect was achieved after 48 h. At the plateau stage, SE-HPS inhibited the proliferation of SCC25 cells more strongly than HPS (P < 0.05). The results of flow cytometry showed that 50 μg/mL HPS and SE-HPS acted on SCC25 cells for 48 h, and the apoptotic rates were 25.8% and 30.8% respectively. Compared with the control group (0 μg/mL HPS and SE-HPS), the difference was statistically significant (P < 0.05). RT-qPCR and Western blotting showed that 50 μg/mL HPS and SE-HPS acted on SCC25 cells for 48 h, and the mRNA and protein expression levels of the apoptosis gene Fas/FasL were upregulated. The difference was statistically significant (P < 0.05).@*Conclusion@# Both HPS and SE-HPS can inhibit the proliferation of SCC25 oral cancer cells, but SE-HPS is superior to HPS and can induce apoptosis through the Fas/Fasl pathway.

Effect of 3 neuroprotective measures on expressions of Fas/FasL in brain tissue of neonatal rats with hypoxicischemic brain damage / 中国中西医结合急救杂志

Objective To observe the effects of 3 neuroprotective measures on the expressions of apoptosis-related factors and their ligands (Fas and FasL) in brain tissue of neonatal rats with hypoxic ischemic brain injury. Methods One hundred and twenty Wistar rats 7 days old were selected as experimental subjects, the rats were divided into four groups: neural stem cell, erythropoietin (EPO), ω-3 unsaturated fatty acid treatment groups and hypoxic ischemic brain damage model group according to random number table method, with 30 rats in each group. Neural stem cell group, EPO group and ω-3 unsaturated fatty acid group were respectively injected with neural stem cells, EPO and ω-3 unsaturated fatty acid, each 5 mL via tail vein after modeling; the hypoxic ischemic brain damage model group was given equal volume of normal saline. At 6, 12, 24, 48 and 72 hours after administration of drug, 6 rats were sacrificed in each group, brain tissue was taken, the mRNA expression levels of Fas/FasL, protein expression levels of Toll-like receptor 4 (TLR4), nuclear transcription factor-κB (NF-κB), tumor necrosis factor-α (TNF-α), interleukin (IL-1β, IL-6) and cell apoptotic rate in hippocampus tissue were measured. Results ① mRNA expressions: the mRNA expressions of Fas and FasL of the 3 experimental groups were significantly lower than those of the hypoxic ischemic brain damage model group, the degrees of descent after administration for 24 hours were the most significant, neural stem cell treatment group < EPO treatment group < ω-3 unsaturated fatty acid treatment group < hypoxic ischemic brain damage model group [Fas mRNA expression (2-ΔΔCt): 140.5±2.9, 156.4±2.5, 165.2±2.7 vs. 173.7±2.8, FasL mRNA expression (2-ΔΔCt): 143.1±4.3, 154.6±1.5, 160.7±1.4 vs. 174.7±2.8], the differences were statistically significant (all P < 0.05). ② Protein expressions: the protein expressions of TLR4, NF-κB, TNF-α, IL-1β, IL-6 of the 3 experimental groups were significantly lower than those of the hypoxic ischemic brain damage model group (TLR4/GAPDH: 0.7±0.2, 0.6±0.1, 0.2±0.1 vs. 1.4±0.1; NF-κB/GAPDH: 6.7±0.4, 5.3±0.1, 1.1±0.2 vs. 11.2±0.3; TNF-α/GAPDH: 14.3±1.4, 11.2±1.2, 3.2±2.1 vs. 23.2±0.5; IL-1β/GAPDH: 9.4±0.2, 7.4±0.3, 2.2±0.3 vs. 13.4±0.1; IL-6/GAPDH: 36.2±4.4, 39.3±1.5, 26.2±2.1 vs. 51.4±1.4, all P < 0.05), the protein expression levels of above indexes in neural stem cell treatment group < those of EPO treatment group < those of ω-3 unsaturated fatty acid treatment group < those of hypoxic ischemic brain damage model group. ③ Apoptotic rates:after drug administration, the apoptotic rates of the ω-3 unsaturated fatty acid group, EPO treatment group, neural stem cell treatment group were obviously lower than the rate of model group [(3.7±0.3)%, (3.4±0.2)%, (2.5±0.1)% vs. (5.5±0.4)%, all P < 0.05]. Conclusion The mRNA expressions of Fas/FasL in the brain of neonatal rats with hypoxic-ischemic brain damage are high, and the treatment with each of the following agents; neural stem cells, EPO and ω-3 unsaturated fatty acid can reduce the mRNA expressions of Fas/FasL in such rats' brain tissues.

Effect of Tripteryginum wilfordii polyglycoside on Fas/FasL and p38 MAPK signaling pathway expression in ulcerative colitis rats / 中国药理学通报

Aim: To investigate the expression changes of Fas/FasL and its related signaling pathway p38 MAPK in ulcerative colitis(UC) rats and the effects of Tripterygium wilfordii polycoride (TWP). Methods: TNBS and ethanol enema method were adopted to build the UC rat model, and colon inflammation in rats was assessed; the differential expression of Fas/FasL was analyzed by expression profiling method, and verified by real-time PCR. Subsequently, pathway analysis in DAVID database showed that Fas/FasL might involve p38 MAPK signaling pathway in regulating UC inflammatory activities, and the related molecules in this signaling pathway were analyzed by the expression profiling method and verified by real-time PCR. Finally, the expression of the terminal inflammatory factors of the pathway were detected by RT-PCR and Western blot. Results: In UC rats, the gross morphological damage and histopathological injury scores significantly increased, which could be reduced by TWP. The trend of FasL was in consistent with that of MAPK14 (p38α), TNF-α and IL-1β in Fas/FasL system and the p38 MAPK signaling pathway, and the expression of them were up-regulated in inflammatory activity, which can be down-regulated by TWP. Conclusion: TWP can regulate the inflammatory activity of UC through Fas/FasL system and the p38 MAPK signaling pathway.

Xanthotoxin induces apoptosis in SGC-7901 cells through death receptor pathway / 中草药·英文版

Objective: To investigate the effects of xanthotoxin from Apiaceae medicinal plants on cell proliferation and apoptosis, and explore its mechanism of action against human gastric carcinoma SGC-7901 cells in vitro. Methods: SGC-7901, HepG-2, MCF-7, and A549 cells were treated with different concentrations of xanthotoxin (10, 20, 60, 80, 100, 120, 140, and 160 µg/mL) for 48 h, and the cell viability (IC50) was determined by MTT assay; Xanthotoxin-induced apoptosis in cells was observed by using Hoechst 33258 Staining Kit and Annexin V-FITC Apoptosis Detection Kit; Flow cytometry was used to detect apoptosis related proteins of Fas/FasL, Bid, and DR5/TRAIL proteins in human gastric carcinoma SGC-7901 cells after being treated by xanthotoxin; The influence of xanthotoxin on Caspase-8 protein expression in the cells was determined by Flouormetric Assay Kit. Results: Xanthotoxin obviously inhibited SGC-7901, HepG-2, MCF-7, and A549 cells proliferation, and its inhibition was in a concentration-dependent manner; flow cytometry results showed that in a certain concentration range, xanthotoxin can increase the expression levels of Fas/FasL and DR5/TRAIL proteins in a concentration-dependence manner. The content of Bid protein in cells was increased, and it showed concentration-dependence. Conclusion: Xanthotoxin may induce SGC-7901 cells apoptosis in a certain concentration range through the Fas/FasL protein mediated death receptor pathway, or by DR5/TRAIL mediated death receptor pathway, and increase the expression level of death receptor protein, activation Caspase-8, activating downstream effect factor, inducing cell apoptosis, or activate Caspase-8 cutting activate protein Bid, and then enter the mitochondrial pathway, induction of apoptosis.

Icariin induces apoptosis of esophageal cancer cell through enhancing Fas-FasL expression activity / 重庆医学

Objective To investigate the inhibitory effect of icariin(ICA) on the xenograft tumors growth of esophageal car‐cinoma and to preliminarily investigate its mechanism .Methods The MTT assay and Giemsa staining were applied to detect and observe the in vitro inhibitory effect of ICA on esophagus cancer cell lines Eca‐109 and TE‐13 .The xenograft tumor model of nude mouse esophagus cancer cell was constructed and divided into 3 groups ,6 cases in each group .Each mouse in the experimental group was intraperitoneally injected by ICA 50 mg/kg ;while the control group was injected by the same volume of normal saline and the positive control group was injected by cis‐platinum 2 mg/kg ,once every 2 days ,a total of 14 days .The tumor volume was measured once per 3 d .After experiment ,the tumor weight was measured;the TUNEL staining was used to observe the morphological chan‐ges and cell apoptosis of tumor tissue in each group .The changes of Fas and FasL protein expression in tumor tissues were analyzed by immunohistochemistry .The FasL and IFN‐γlevels in peripheral blood were tested by the ELISA assay .Results ICA exerted no obvious inhibitory effect on the proliferation of Eca‐109 and TE‐13 cell in vitro .The average volume and weight of xenografts tumor had statistical difference between the experimental group and the positive control group (P<0 .05) .The TUNEL staining results showed that the tumor tissues had obvious apoptosis ,the number of apoptosis cells was significantly increased compared with the control group(P<0 .05) .The immunohistochemistry experimental results showed that the expression of Fas and FasL was signifi‐cantly increased(P<0 .05) .The ELISA experimental results demonstrated that the FasL and IFN‐γlevels of peripheral blood in the experimental group were significantly increased(P<0 .05) .Conclusion ICA had no obvious inhibitory effect on esophageal cancer cell proliferation in vitro ,but could induce in vivo apoptosis through the Fas expression and secretion of FasL and IFN‐γ,thus plays the role of anti‐esophageal cancer .

Study on role of Fas/FasL in pathogenesis of primary immune thrombocytopenia / 重庆医学

Objective To explore the role of Fas and FasL system in the pathogenesis of adult primary immune thrombocyto‐penic purpura (ITP) .Methods The peripheral anticoagulant venous blood samples were collected from the patients with newly di‐agnosed ITP and healthy adults .The expression rates of Fas and FasL in Th/Tc ,Th1/Th2 ,Tc1/Tc2 cells and their expression rates at platelet surface were detected by flow cytometry .Results The Fas and FasL expression rates on the surface of Th ,Th1 ,Th2 , Tc ,Tc1 and Tc2 in the ITP group were increased compared with the healthy control group(P0 .05) .Conclusion The Fas and FasL expression on T cell subsets and platelet surface in ITP patients is abnormal , which may be related with the pathogenesis of ITP .

Roles and mechanisms of Fas/FasL in the apoptosis of HL-60 cells induced by euphorbiasteroid / 国际肿瘤学杂志

Objective To investigate the effect of euphorbiasteroid on inducing the apoptosis of HL-60 cells and demonstrate whether the Fas/FasL signaling pathway is involved in the induction of apoptosis. Methods HL-60 cells were treated with dose of 2.5,10,40 μg/ml of euphorbiasteroid in vitro for 24 h respectively.After that,cell counting Kit-8 was used to detect cell proliferation.The morphology of HL-60 cells were observed under light and fluorescent microscopy.The early cell apoptosis was detected by using flow cytometry with Annexin Ⅴ-FITC /PI double staining.The expressions of Fas,FasL,caspase-8 and caspase-3 mRNA were analyzed by the method of RT-PCR.The activities of caspase-8 and caspase-3 were examined by chromatometry.Results Compared with 1640 control group,HL-60 cell proliferation was inhibited significant-ly by euphorbiasteroid.The inhibition rates were (34.9 ±3.7)%,(54.6 ±5.2)% and (61.3 ±4.3)%respectively.Moreover,HL-60 cells exhibited typical morphological features.Early cell apoptosis rates of HL-60 cells were (23.4 ±3.1)%,(35.7 ±4.3)% and (53.2 ±3.9)% respectively.Furthermore,the expressions of Fas,FasL,caspase-3 and caspase-8 mRNA were up-regulated significantly after euphorbiasteroid administration in a dose-dependent manner (P<0.01 ).After treated with euphorbiasteroid,the activities of caspase-8 and caspase-3 were significantly enhanced (P<0.01 ).Conclusion The up-regulation effect of euphorbiasteroid on Fas/FasL signaling pathway might contribute to the apoptosis of HL-60 cells.

Lipopolysaccharide and outer membrane proteins of Leptospira interrogans induce macrophage apop-tosis via Fas/FasL pathway / 中华微生物学和免疫学杂志

Objective To investigate the role of pathogenic Leptospira interrogans lipopolysaccha-ride (L-LPS) and outer membrane proteins (L-OMP) in the apoptosis of mouse macrophages (J774A.1) and their association with Fas/FasL pathway .Methods Phenol-water extraction and Triton X-114 phase separation were used to extract L-LPS and L-OMP from L.interrogans serogroup icterohaemorrhagiae serovar Lai strain Lai, respectively.Polymyxin B ( PMB) and protease K ( PK) were used to treat L-LPS and L-OMP, respectively.J774A.1 cells were stimulated by L.interrogans strain Lai with or without ultraviolet inactivation.In parallel, the cells were stimulated by extracted L-LPS and L-OMP with or without PMB and PK treatments .The apoptosis and necrosis of J 774 A.1 cells before and after treatment were detected by flow cytometry.The siRNAs were used to silence the expression of Fas or FasL gene in J 774A.1 cells and their inhibitory effects were further validated by using real-time fluorescent quantitative RT-PCR.Flow cytometry was used to detect the effects of L-LPS or L-OMP on the apoptosis of J774A.1 cells with Fas or FasL gene-knockdown .Results L.interrogans strain Lai with or without ultraviolet inactivation could cause similar early apoptosis rates (47.1%and 55.6%) and late apoptosis/necrosis rates (7.6%and 7.9%).The ear-ly apoptosis rates of 1×105 J774A.1 cells were 40.4%and 34.0%after the treatment with 100 ng of L-LPS and 100 μg of L-OMP for 4 h.The late apoptosis/necrosis rates of the cells were 7.5%and 6.9%upon the treatments with L-LPS and L-OMP, respectively.However, the apoptosis or necrosis of the cells was not ob-served when using L-LPS and L-OMP pre-treated by PMB and PK, respectively.Silenced expression of Fas or FasL gene reduced the L-LPS-induced J774A.1 cells apoptosis (P<0.05), while decreased early apopto-sis rate of J774A.1 cells mediated by L-OMP was only observed in Fas gene-knockdown cells (P<0.05). Conclusion Both L-LPS and L-OMP can cause the Fas/FasL-associated apoptosis of macrophages , which is beneficial for L.interrogans to establish the productive infection in hosts .

Change of FAS/FASL System after Unilateral Testicular Torsion in SD Rats at Puberty / 昆明医科大学学报

Objective To investigate the relationship between the expression of FAS/FASL and spermatogenesis in the contralateral testis after unilateral testicular torsion in puberty Sprague-Dawley rats. Methods Five groups of side testicular torsion adolescence SD rats model were established as follows:group A as control group, group B for testicular torsion group,group C for testicular torsion plus methylprednisolone group, D for reverse side testes excision group, and Group E testes excision plus methylprednisolone group for reverse side. SD rats were treated with testicular resection and/or injection of methylprednisolone after torsion for 24 h. The rats were executed at postoperative 1 month, and contralateral testes were collected for histopathological examination. Expression of FAS and FASL was analyzed quantitatively by immunohistochemistry with a computer pathological image analysis system. Each rat was evaluated with regard to endocrine parameters (follicle-stimulating hormone (FSH), luteinizing hormone ( LH) and testosterone ( T) by radioimmunoassay. Results Histopathological examination of the contralateral testes showed that either orchiectomy or orchiectomy plus methylprednisolone was more successful than no treatment. Fas/FasL protein in both control group and 4 experimental groups were expressed. B group showed more expression of FAS and FASL than that of the other groups. FSH, LH and T were normal in all cases. Conclusion Unilateral testicular torsion in puberty SD rats may result in higher expression of FAS and FASL, and accelerate germ cell apoptosis and subfertility. Methylprednisolone may decrease the expression of FAS and FASL and maintain spermatogenesis in the contralateral testis after the unilateral testicular torsion.

Fas and FasL expression in leukocytes from Chronic Granulomatous Disease patients / Expresión de Fas y FasL en leucocitos de pacientes con Enfermedad Granulomatosa Crónica

Chronic Granulomatous Disease (CGD) is a primary immunodeficiency characterized by defects in superoxide (O2-) production, which result from mutations in one of the four NADPH oxidase components, predisposing to bacterial and fungal infections. Besides the O2-defect, it has been described that neutrophils from CGD patients are resistant to cell death, a phenomenon that has been connected to chronic inflammation and predisposition to autoimmune diseases. A diminished expression of Fas and its counterpart FasL, molecules known to play a major role in cell death, has been described in lymphocytes depleted of O2-reactive oxygen species (ROS), suggesting an involvement of ROS in Fas/FasL expression. In this work, Fas and FasL expressions were analyzed in T cells and neutrophils from two CGD families, previously known to harbor two different molecular defects: absence of either p47-phox or p67-phox. We found that T lymphocytes from CGD patients express low levels of Fas and FasL, while a diminished FasL expression was observed on neutrophils from a CGD A470 patient. These defects may contribute to understand altered cell death in CGD patients.

La Enfermedad Granulomatosa Crónica (EGC) es una inmunodeficiencia primaria caracterizada por un defecto en la producción de superóxido (O2-), que se genera como consecuencia de mutaciones en uno de los cuatro componentes del complejo NADPH oxidasa y predispone a infecciones por bacterias y hongos. Además de los defectos en la producción de O2-, se ha descrito que los neutrófilos de los pacientes con EGC exhiben una resistencia a la muerte celular, evento que se asocia con la inflamación crónica y predisposición a enfermedades autoinmunes. Se ha descrito que linfocitos en medios desprovistos de O2-especies reactivas del oxigeno (ROS), muestran reducida expresión de Fas y FasL, moléculas que juegan un papel relevante en el control de la muerte celular, sugiriendo la participación de los ROS su regulación. En este trabajo analizamos la expresión de Fas y FasL en linfocitos T y neutrófilos en dos familias portadores de dos defectos genéticos diferentes asociados con EGC: ausencia de p47-phox o de p67-phox. Evidenciamos una baja expresión de Fas y FasL en los linfocitos T de los pacientes con EGC, pero solo los neutrófilos de los pacientes con defecto de p47-phox, fueron incapaces de expresar FasL. Estos defectos pudieran contribuir a entender la alteración de la muerte celular observada en los pacientes con EGC.

Quinazolinone exposure induces apoptosis and changes expressions of Fas/FasL and C-Flip in embryonic mice testicles.

Quinazolinones represent a class of sedative and anticancer drugs. Quinazolinones-based compounds have ability to suppress prostate tumor growth via apoptosis. Apoptosis is very common in embryos and adults of normal and injured mammalian testes. Effects a new derivative of quinazolinone (4(3H) quinazolinone-2-ethyl-2-phenyl ethyl (QEPE)), on the testis of Balb/C mice embryos were investigated. QEPE was able to reduce number of germ cells and diameter of seminiferous tubules. TUNEL assay analysis indicated that reduction correlated with an increase in the number of apoptotic cell. Furthermore, electron microscope observations confirmed typical apoptotic morphologies characterized by chromatin fragmentation. Finally, RT–PCR analysis showed QEPE increases the levels of Fas/Fasl and decreases C-Flip mRNAs in the testis of exposed embryos.

Study on the isoniazid induced cellular damage and the expressions of Fas/Fas ligand of HepG2 / 中华传染病杂志

Objective To establish a model of isoniazid induced necrosis and apoptosis in HepG2 cell and to observe the expressions of Fas/Fas ligand (FasL) in this model.Methods HepG2 cells were treated with different dosages of isoniazid (0,1,2,4,6 and 8 mg/mL) or blank control for 24 hours.Flow cytometer was used to observe the cellular morphology of the HepG2 cell.Annexin V/propidium iodide double staining and flow cytometry were employed to detect the necrosis and apoptosis of HepG2 cells.The expressions of Fas/FasL on the cells were also determined by flow cytometry.The data were analyzed by one-way ANOVA.The comparisons between the drug groups and the control group were performed by using Dunnett t test. Results The higher the dose of isoniazid (4,6,8 mg/mL) was,the more necrosis and apoptosis were observed.In the 4,6 and 8mg/mL isoniazid arms,the total mortality rates were all higher than the control group [(32.1 ±7.5)％,(34.9±8.1)％,(38.2±9.4)％ vs (7.2±1.5)％ respectively](t=4.62,5.14 and 5.75,respectively; all P＜0.01 ).The expression levels of Fas increased along with the dose of isoniazid increasing [(8.7±2.2)％,(11.5±2.8)％,(12.3±3.0)％ and (10.6±2.9)％ in isoniazid 2,4,6and 8 mg/mL arms,respectively],which were all higher than that in control arm [(3.1 ±0.8) ％](t=2.97,P＜0.05; t=4.46,P＜0.01; t=4.88,P＜0.01; t=3.98,P＜0.05).Furthermore,the expressions of FasL increased as well when the dose of isoniazid increased.The expression levels of FasL were (16.2±3.5)％,(21.7±4.8)％ and (18.7±4.9)％,respectively in isoniazid 4,6 and 8 mg/mL arms,which were all higher than that in the control group [(7.4±1.4)％](t=3.11,P＜0.01; t=5.06,P＜0.01; t=3.99,P＜0.05).HepG2 cell necrosis increased with isoniazid of 8 mg/mL.However,the increase of apoptosis was not observed.Conclusion Isoniazid can induce HepG2 cell necrosis and apoptosis,and the apoptosis may be related with the increased expressions of the Fas/FasL on the cells.

J774A. 1 cell apoptosis induced by Leptospira interrogans lipopolysaccharide and apoptotic regulation of associated signaling pathways / 中华微生物学和免疫学杂志

Objective To determine the effect of Leptospira interrogans lipopolysaccharide (L-LPS) inducing apoptosis of murine mononuclear-macrophage cell line( J774A. 1 ), and apoptotic regulation of Toll-like receptor(TLR) and associated intracellular signaling pathways. Methods Lipopolysaccharide (L-LPS) of L. interrogans serogroup Icterohaemorrhagiae serovar Lai strain Lai 56601 was prepared using phenol-water method. The effects of L-LPS inducing J774A. 1 cell apoptosis and the apoptosis-blocking with FasL neutralizing antibody were detected by flow cytometry. Real-time fluorescent quantitative RT-PCR (qPCR) and flow cytometry were performed to measure the changes of Fas/FasL mRNA and protein expression levels in J774A. 1 cells before and after L-LPS treatment. The regulations in L-LPS-induced cell apoptosis by TLR2 and TLR4 as well as p38MAPK, JNK, ERK pathways were determined by either TLR2 or TLR4 antibody blocking test, signaling pathway blocking test and flow cytometry. Results 56.50%, 69.28% and 24.35% of the J774A. 1 cells after treatment with 100 ng/ml L-LPS for4, 12 and 24 h were apoptotic,while the apoptosis rates were decreased to 11.21%, 21.58% and 12.70% after the cells blocked by FasL neutralizing antibody(P ＜0.05). The levels of FasL and Fas mRNAs in J774A. 1 cells treated with L-LPS for 4, 12 and 24 h were elevated with 1.34, 2.12, 2.10 times and 2.45, 3.87, 3.12 times compared to those in the L-LPS untreated cells (P ＜ 0. 05 ), respectively, while the expression rates of FasL and Fas proteins were upregulated to 18.61%, 60.13%, 42.75% and 76.34%, 85.70%, 77.92% from 4.82% and 15.32% apoptotic rates in the L-LPS untreated cells, respectively( P ＜0.05 ). The L-LPS-induced apoptosis rate( 11.54% ) of TLR2 antibody blocked J774A. 1 cells was significantly lower than that(66.56% ) of the J774A. 1 cells without TLR2 antibody blocking( P ＜0.05 ), but L-LPS-induced apoptosis rate of TLR4 antibody blocked J774A. 1 cells was as high as 55.27% ( P ＞ 0.05 ). Compared to the apoptosis rate (62.17%) in the p38MAPK and JNK pathway-free J774A. 1 cells, the L-LPS-induced apoptosis rates in p38MAPK blocked cells(20.54% ) and JNK blocked cells(47.98% ) were significantly lower( P ＜0.05 ),and the apoptosis rate in ERK blocked cells was as high as 61.72% ( P ＞ 0.05 ). Conclusion L-LPS was recognized by TLR2 and upregulates both Fas and FasL expression via p38MAPK and JNK pathways, which involving in the process of the L-LPS-induced cell apoptosis.

Effect of arsenic trioxide in inducing apoptosis and expression of Fas/FasL of gastric tumor cells in nude mice / 西安交通大学学报(医学版)

Objective To study the effect of arsenic trioxide (As_2O_3) on the expression of Fas/FasL and in inducing apoptosis of gastric tumor cells in nude mice. Methods The xenograft model was established in 30 male BALB/C-nu/nu mice. The mice were randomly divided into 3 groups and injected intraperitoneally with As_2O_3 (2.5mg/kg or 5mg/kg) or with saline solution of the same volume respectively. Tumor growth was detected by measuring tumor volume. Apoptosis of the tumor cells was measured with transmission electron microscope (TEM) and TdT-mediated dUTP nick end labeling (TUNEL). The expression of Fas/FasL in the tumor cells was detected with immunohistochemistry. Results Tumor volume in As_2O_3-treated groups was smaller than that in control group (P<0.05). Apoptotic cells in the As_2O_3-treated groups significantly outnumbered than those in control group (P<0.01). The expression of Fas in tumor cells was higher in the As_2O_3-treated groups than in control group (P<0.05), but FasL expression of tumor cells did not differ between the treated groups and control group (P>0.05). Conclusion As_2O_3 can obviously inhibit the growth of human gastric tumor. One of the mechanisms is up-regulation of Fas gene that can induce the apoptosis of gastric cells.

Apoptosis of K-562 cells induced by oleum curcumae wenchowensis and its mechanisms / 中国生化药物杂志

Purpose To investigate the effects and the mechanism of oleum curcumae wenchowensis (OCW) with different concentrations (0, 2.5, 5, 10 and 20 mg/mL) on chronic myeloid leukemia cell line K-562 cells in vitro.Methods The apoptosis of K-562 cells was dyed by Hoechest 33258 and detected by flow cytometry marked with Annexin V/PI. The Expression of Fas/FasL, bcr/abl, bcl-2 and p53 was detected by semi-quantity reverse transcript-polymerase chain reaction (RT-PCR) and Western blot.Results The results showed that the apoptosis rates were gradually elevated. The expression of Fas and FasL protein was increased in a concentration dependent manner, while bcr/abl, bcl-2 and p53 had no significant changes.Conclusion OCW could induce the apoptosis of K-562 cells by up-regulating the expression of Fas/FasL protein.

Effects of somatostatin on the apoptosis of colorectal cancer cells / 中华消化外科杂志

Objective To investigate the effects of somatostatin on the apoptosis of colorectal cancer cells. Methods The expression of somatostatin mRNA in colorectal cancer tissues from 79 patients who had been admired to Yijishan Hospital from January 2004 to October 2006 was detected by nested RT-PCR. The apoptotic index of colorectal cancer cells was detected by TUNEL, and the protein expressions of somatostatin, Fas, FasL, caspase-3 and caspase-8 in colorectal cancer tissues were detected by immunohistochemistry. All data were analyzed by chi-square test, q test and Spearman rank correlation coefficient. Results There was a positive correlation between the mRNA and protein expression of somatostatin (r = 0.98, P < 0.05). The mRNA and protein expression of somatostatin in poorly and moderately differentiated colorectal cancers were significantly lower than that in well differentiated colorectal cancers (χ~2 = 10.78, 11.24, 5.27, 5.24, P < 0.05). The positive expression rates of mRNA and protein of somatostatin in papillary adenocarcinoma were significantly higher than those in mucinous adenocarcinoma and signet ring cell carcinoma and undifferentiated carcinoma (χ~2= 6.56, 6.99, 5.44, 7.39, P < 0.05). The mRNA and protein expression of somatostatin in colorectal cancer in Dukes A and B were significantly higher than that in Dukes C and D (χ~2 =5.17, 4.06, P <0.05). The apoptotic index in high or moderate somatostatin expression group was significantly higher than that in low somatostain expression group (q = 5.66, 4.21, P < 0.05), and the positive expression rates of Fas, caspase-8 and caspase-3 in high or moderate somatostatin expression group were significantly higher than those in low somatostafin expression group (χ~2= 5.48, 5.62, 6.89, 4.32, 4.19, 3.91, P <0.05). Conclusion Somatostatin plays an important role in the regulation of cell apoptosis in colorectal cancer, and the mechanism may be related to the aberrant expression of Fas/FasL.

The investigation of expression of CD95, CD44V6 in breast cancer and lymph node micrometastasis / 肿瘤研究与临床

Objective To investigate the relationship and clinicopathologic significance between the expression of CD95,CD44V6 and metastasis of axillary lymph node in breast cancer.Methods Immunohistochemistry was used to detect the expression of CD95 and CD44V6 in 101 cases of breast cancer, in which, 40 cases without metastasis in axillary lymph node were detected MCK expression by IHC. The results were analyzed statistically. Results 9 cases of breast cancer with lymph node micrometastasis were observed by IHC in 40 cases without metastasis in axillary lymph node by microscope. The expression of CD95, CD44V6 in lymph node metastasis group was similar as in lymph node micrometastasis group. There was significant difference of CD95 expression between those with lymph node metastasis and those without. The positive rate of CD95 and the high expression of CD44V6 in the cases that the tumor size was over 2 cm were significandy higher than in the cases that the tumor size was less than 2 cm (P<0.05). Conclusion Detection of the expression of CD95, CD44V6 in breast cancer may be helpful to predict the lymph node micrometastasis and provide more dependable evidence for judging prognosis and selecting treatment prescription clinically.

Effects of emodin on expression of PMN Fas/FasL,Caspase-3 from peripheral blood in patients with SIRS / 中华肝胆外科杂志

Objective To investigate the expression of PMN Fas/FasL,Caspase-3 from peripheral blood in patients with SIRS and observe the effect of emodin on the PMN apoptosis and explore its mechanism.Methods The samples of peripheral blood were collected from 8 patients(all suffer from acute pancreatitis)with SIRS and 6 healthy volunteers.The circulating neutrophils were isolated and cultured.There were three groups of in our study:control group,SIRS group and emodin-treated group.The change of the PMN apoptosis and the expression Ieve]of Fas/FasL.Caspas-3 were observed.Results Compared with healthy volunteers.the percentage of PMN apoptosis significantly reduced in patients with S1RS(P＜0.05).After emodin was used,the percentage of PMN apoptosis in patients with SIRS increased(P＜0.05).Compared with healthy volunteers,the expression of Fas and Caspase-3 reduced in patients with SIRS(P＜0.05).Emodin could significantly induce the expression of Fas and Caspase-3(P＜0.05).After 24 hours of in vitro culture.the expression of FasL was not detected.Conclusion PMN from patients with SIRS shows profoundly delayed rates of apoptosis in vitro compared with PMN from healthy volunteers,and the decreased percentage of PMN apoptosis is associated with the reduced expression of Fas and Caspase-3.Emodin can significantly inhibit the delayed PMN apoptosis in patients with SIRS by inducing the expression of Fas and Caspase-3.