Influence of Moxibustion Serum on the Expression of Fas bcl-2 mRNA and Protein of EL-4 Lymphoma Cells / 针灸推拿医学（英文版）

Objective: To observe the influence of moxibustion sertma of mice on the expression of Fas, bcl-2 mRNA and protein of EL-4 lymphoma cells in vitro. Method: The EL-4 lymphoma cells were cultivated for 24 h in the moxibustion serum of mice. The expression of Fas and bcl-2 mRNA of EL-4 lymphoma cells were detected by in-situ hybridization method, and the protein expression of Fas and bcl-2 were observed by the immuocytochemistry method. Results: The expression of bcl-2 mRNA and protein decreased, and the expression of Fas rnRNA and protein increased significantly in EL-4 cells, which were cultivated in the moxibustion serum compared those cultivated in normal mice serum (P<0.05). Conclusion: Moxibustion serum could down-regulate the bcl-2 mRNA and protein and up-regulate the Fas mRNA and protein of EL-4 cells.

The Expression of Fas antigen and Bax and Apoptosis in Ex Vivo Expanded Hematopoietic Progenitor Cells / 대한혈액학회지

BACKGROUND: During ex vivo expansion of cord blood (CB) CD34+ cells, differentiation of the expanded cells happened and hematopoietic potential of the progenitor cells decreased. In this study, we evaluate the effect of the expression of Fas antigen, Bcl-2, and Bax on CD34+ or AC133+ hematopoietic progenitor cells during ex vivo expansion. METHODS: CD34+ and AC133+ cells isolated from human CB were cultured in serum free medium supplemented with several cytokines for 7 days. After expansion culture, we re isolated CD34+ and AC133+ cells and compared the numbers of granulocyte-macrophage colony-forming units (CFU-GM) and granulocyte, erythrocyte, monocyte, and macrophage colony-forming units (CFU-GEMM), and expression of Fas antigen, Bcl-2, and Bax with unexpanded cells. RESULTS: CFU-GM was expanded 23.94 fold in CD34+ cells and 15.22 fold in AC133+ cells at day 7 of culture but CFU-GEMM was not expanded. The expression of Fas antigen and Bax was 7.44% and 2.75%, respectively, in fresh isolated CD34+ cells and increased to 19.71 % and 33.67%, respectively, in expanded CD34+ cells at day 7 culture, but Bcl-2 was not changed. In case of AC133+ cells, the expression of Fas antigen and Bax were also increased from 5.87% and 6.19% to 24.85% and 22.83%, respectively, and Bcl-2 was slightly decreased. Apoptosis was not changed in CD34+ cells and AC133+ cells during ex vivo expansion. CONCLUSION: These results indicate that the nature of expansion was similar between CD34+ and AC133+ cells, and expression of Fas antigen and Bax increased on CD34+ and AC133+ cells during ex vivo expansion. Selection of the expanded progenitor cells without apoptosis may be useful for the hematopoietic stem cell transplantation.

Increased Expression of Fas Antigen and Apoptosis in Aplastic Anemia Bone Marrow Cells

BACKGROUND: Clinical observations and laboratory studies have supported an immune basis for most acquired aplastic anemias, with the majority of patients responding to immunosuppressive therapy. Fas, a member of the tumor necrosis factor (TNF) receptor superfamily is a critical downregulator of cellular immune responses. Proinflammatory cytokines like interferon gamma (IFN-gamma) and TNF-alpha can induce Fas expression and render hematopoietic progenitor cells susceptible to Fas-induced growth suppression and apoptosis. METHODS: In order to investigate the involvement of apoptosis in the pathogenesis of aplastic anemia (AA), we measured the expression of Fas antigen and caspase-3 on bone marrow (BM) mononuclear cells (MNCs) of AA in the presence or absence of IFN-gamma, TNF-alpha, or macrophage inflammatory protein 1-alpha (MIP-1alpha). RESULTS: We confirmed that AA BM MNCs were more apoptotic and highly expressed Fas antigen than normal donors. Stimulation by IFN-gamma, TNF-alpha, or MIP-1alpha increased Fas antigen and caspase-3 expression in AA BM MNCs than BM MNCs of normal donors. Anti-Fas monoclonal antibody enhanced IFN-gamma, TNF-alpha, or MIP-1alpha mediated caspase-3 expression in BM MNCs of normal donors. Among these three cytokines, IFN-gamma enhanced apoptosis most strongly via Fas-caspase-3 pathway. CONCLUSION: These results suggest that Fas signal pathway may play a role in the pathophysiology of aplastic anemia and negative hematopoietic regulators like IFN-gamma can induce apoptosis of bone marrow progenitors in part by Fas induction.

Induction of Fas Antigen Expression, and Apotopsis of Human B Cell by CD40 and Tumor Necrosis Factor Receptor Ligation / 대한류마티스학회지

OBJECTIVE: CD40 and the TNFR belong to the NGF/TNFR supergene family. Ligation of CD40 on B cells induces activation ignals leading to proliferation, Ig isotype class switching, germinal center formation but also induces Fas antigen expression.In addition,CD40 ligation induces pro-inflammatory cytokines including TNF-alpha and LT-alpha gene transcription by human B cell.TNF-alpha is a pleiotropic cytokine and also induces Fas antigen expression on various cells. Lately it has been known that TNF-alpha plays an important role in the pathogenesis of chronic inflammatory diseases,including rheumatoid arthritis,or chronic inflammatory bowel diseases.However there have been occurrence of autoantibodies,or autoimmune disease such as lupus after use of anti TNF-alpha blocking agents. In this report,we tested the relationship and biological significance of CD40 ligation and TNFR signaling with respect to Fas antigen expression on human B cells. METHODS: Ramos Burkitt's lymphoma B cell was used as a prototype of ger-minal center B lymphocyte,and R2G6 cell was utilized as a model of activated germinal center B cell.CD40 lgation was performed by the coculture with CD40 ligand bearing L-293 cells,or anti-CD40 monoclonal antibody,whereas control was obtained with CD-8-L-293 cells or control antibody.Expression of Fas antigen was determined with flow cytometer.Apoptosis assay was conducted by two ways.Alamar blue reduction assay after sIgM cross linking or anti-Fas anti-body,in the presence or absence of CD40 ligation or TNF-alpha .In addition,DNA content assay was utilized to make sure the proportion of apoptotic Ramos B cells by various treatments. RESULTS: 1)CD40 and TNF-alpha induced Fas antigen expression on Ramos B cell line cells and rendered them susceptible to Fas-mediated apoptosis.2)CD40 and TNFR signaling upregulate Fas antigen independently.3)Both TNFR and CD40 signaling rescue sIgM crosslink induced apoptosis of Ramos B cell line cells,only CD40,but not TNFR,signaling rescues Ramos cells from Fas-mediated apoptosis. CONCLUSION: Taken together,these results demonstrate that B cell signaling via two distinct members of the NGF/TNFR superfamily,CD40 and TNFR, independently engage the Fas pathway and provide mechanisms for eliminating B cells.Acting alone,both signals will ready B cells for Fas-mediated apoptosis. In concert with sIg signaling,the rescue effect provided uniquely by CD40 ligation assures the selective survival of only those B cells which have bound antigen and presented it to antigen-specific T(h) cells .

The Expression of Fas Antigen and Apoptosis in Patients with Myelodysplastic Syndromes and Its Clinical Relevance / 대한혈액학회지

BACKGROUND: The myelodysplastic syndromes (MDS) are a group of acquired clonal hematopoietic disorders characterized by the peripheral cytopenias and hypercellular or normocellular dysplastic bone marrow. The event responsible for the development of MDS is generally not known. Several recent reports have described an increased frequency of apoptosis in bone marrow cells from patients with aplastic anemia or MDS. Gersuk et al observed that Fas ligand expression was significantly increased on bone marrow cells from MDS patients as compared to normal individuals. METHODS: We examined apoptosis and Fas antigen expression using ISNEL method and immunohistochemistry on marrow cells from 36 patients with MDS. RESULTS: Among the 36 patients, 15 patients (42%) demonstrated apoptosis positive cells (>15%) and 9 patients (25%) demonstrated positive Fas antigen expression (>20%). The presence of apoptosis significantly correlated with hemoglobin level at diagnosis (P<0.05) and the expression of Fas antigen significantly correlated with bone marrow cellularity and CD34 positive cell aggregate group at diagnosis (P<0.05). There was no statistically significant relationship between apoptosis and Fas antigen expression. The median survival of all patients with MDS was 44 months (2-117+). Multivariate analysis showed that FAB classification and hemoglobin level at diagnosis were significant prognostic factor but presence of apoptosis and expression of Fas antigen had no significance. CONCLUSION: The data indicate that the apoptosis and expression of Fas antigen are present in patients with MDS and correlate with some clinical parameters but not significantly associated with survival period of the patients with MDS.

APOPTOSIS INDUCED BY CYCLOSPORINE A IN RENAL TUBULAR AND INTERSTITIAL CELLS AND THE PROTECTIVE EFFECTS OF ENALAPRIL AND LOSARTAN ON IT / 解放军医学杂志

The rats were fed on low salt diet for seven days,and then were randomly divided into five groups: control (n=7), CsA treated (n=7), CsA+Verapamil (n=7), CsA+Enalapril (n=7) and CsA+Losartan (n=7).CsA was given by subcutaneous injection (15mg?kg -1?d -1) for four weeks. The apoptosis of tubular and interstitial cells was detected by TUNEL assay. The proliferating cell nuclear antigen and Fas antigen expression were examined by immunohistochemical staining. Apoptosis of tubular and interstitial cells increased in CsA treated rats. CsA+Losartan and CsA+Enalapril treated rats had a statistically significant decrease in apoptosis when compared to CsA treated rats (7.3?1.1 and 6.6?0.8 vs 13.9?1.2 respectively, P