Effects of radiation on nuclear factor-kappa B in bone marrow stromal cells
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Objective To study the changes of the microenvironment of bone marrow hematopiesis after exposure to gamma radiation. Methods Immunohistochemistry and electrophoretic mobility shift assay (EMSA) were used. Results The expression of nuclear factor kappa B(NFκB) protein in the bone marrow stromal cells (BMSCs) was elevated after exposure to 8 Gy gamma rays radiation determined with immunohistochemistry. The activity of NFκB in the BMSCs was significantly increased after irradia tion determined with EMSA and it reached the peak 4 hours after irradiation. Conclusion Our findings suggest that nuclear factor kappa B in the BMSCs is involved in the protection of BMSCs and in the reco very of hematopoiesis after exposure to radiation.Cite
Objective To explore the change of inhibitors expression by bone marrow stromal cells(BMC S)after radiation injury in mice.Methods The mouse model of radiation injury was repared by radiating mice by 6GyCo 60 ?TNF α?TGF β 1and MIP 1αwas detected by radioimmunoassay and ELISA methods respectively,at same time the relationship between the levels of inhibitors and the quanitity of hydrocortisone and numbers of bone marrow cell(BMSC S)was showed. Results The levels of TNF α?TGF β 1 and MIP 1α was dramaticly increased in 3d?7d,in BMSC S culture medium after radiation injury,hydrocortisone can obviously inhibit the expression of inhibitors,levels of inhibitors was markedly related to the numbers of BMCS. Conclusion The disfunction of hematopoiesis after radiation injury was related to the increased levels of inhibitors,hydrocortisone has obvious suppression effect on expression of inhibitors.
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In order to investigate focal flushing dose radiation effect on the generation, differentiation and gene expression of bone marrow stromal cells the femoral head of rats was irradiated at 30 Gy by 137Cs γ-rays (dose rate: 0.83 Gy/min). Then the bone marrow stromal cells (BMSCs) was cultured. The ability of proliferation and colony formation was observed and the expression level of Cbf-α1, PPAR-γ, VEGF-a and KDR was detected by RT-PCR technology in two weeks later. It has been found that after partly irradiated by 137Cs γ-rays the proliferation and the number of colony of BMSCs in irradiated group decreases obviously meanwhile the expression level of Cbf-α1, PPAR-γ and VEGF-a in irradiated BMSCs obviously decreases by 18.98 %, 9.46 %, 57.34 % and 5.56 % respectively compared to the normal BMSCs (p0.05). It shows that the focal great radiation could damage the BMSCs and depress the generation, differentiation and the expression of the related genes obviously.
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Objective To reveal the abnormity of bone marrow microenvironment of psoriasis patients by comparing the level of TNF-α、LIF and HGF secreted by bone marrow stromal cells in patients and control.Methods Separate bone marrow mononuclear cells of patients and control by density gradient centrifugating.Bone marrow stromal cells were cultured with the method of adherent.When the cells were cultured for three passages and 72h,bone marrow stromal cells and supernatant liquid were collected.The phenotypes of cells were identified by flow cytometry and the level of TNF-α,LIF and HGF were detected by ELISA kits.Results The purity of bone marrow stromal cells was over 90%.The level of TNF-α,LIF and HGF in patients was significantly lower than that in control(P0.05).Conclusions Some of the cytokines which secreted by psoriatic bone marrow stromal cells are abnormal.This shows that psoriatic bone marrow microenvironment may be abnormal.
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In order to investigate radiation effect on the expression of VEGF in murine bone marrow stromal cells, C57BL / 6 J mice were irradiated to 0, 2.5, 5 and 10 Gy by whole body irradiation with 137Cs γ?ray. After the bone marrow stromal cells were ex-vivo cultured for 3, 6 or 10 d, the mRNA and protein of VEGF in BMSCs were analyzed by RT?PCR and ELISA, respectively. Both of the two indices were up-regulated with the cultural time in the control group (without irradiation). In the 5 and 10 Gy group, the expressions of VEGF mRNA in BMSCs increased significantly with the cultural time. Interestingly, in the 5 and 10?Gy group, VEGF concentration in BMSCs cultural media increased on 6 d and reduced significantly on 10 d after the ex?vivo culture. This is because that total cell numbers of the 10d group had been reduced significantly in comparison to the control group of the same cultural time. These results indicated the expressions of VEGF in BMSCs at mRNA and protein levels increased in each cell after 5 and 10 Gy radiation. Up-regulated VEGF might benefit for hematopoietic reconstitution. The asso-ciation among expression of VEGF, hematopoietic microenvironment and hematopoietic reconstitution deserve fur-ther study.
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Immunoregulation of bone marrow stromal cells transfected interleukin 18 on intracranial glioma rats
Objective:To clarify the mechanism of immunoregulation of bone marrow stromal cells(BMSCs) transfected interleukin 18(IL-18) after their transplantation into glioma bearing rats.Methods:Cultured BMSCs from SD rats were transfected with rmIL-18(BMSCs/IL-18).Untransfected BMSCs were used as control.Culture supernatant medium was collected for IL-18 examination at different time point by ELISA kit.After establishment of glioma bearing rats followed by BMSCs/IL-18 transplantation,serum concentration of IFN-γ,IL-2 and IL-10 were examined by means of ELISA kit.And their splenocytes were cultured with C6 cells and BMSCs/IL-18 for in vitro cytotoxicity assay,and subsets of splenocyte were detected by flow cytometry.TUNEL was used to clarify apoptosis cells inside glioma and anti-CD34 staining was performed to observe microvessel density(MVD).Results:BMSCs/IL-18 could secret IL-18 long term and stably.After being transplanted with BMSCs/IL-18,serum concentration of IL-2,IFN-γ in glioma bearing rats' increased obviously and serum concentration of IL-10 decreased.Flow cytometry results showed that CD4+ and CD8+ T lymphocytes increased in the splenocytes.And rechallenge with C6 cells induced a rapid immuno-reaction.In vitro cytotoxicity assays.It was showed that BMSCs/IL-18 could stimulate splenocytes to kill C6 cells obviously.TUNEL assay showed that there were 15.74±6.23 apoptosis cells inside glioma in each view in Group 2,which was much more when compared with other groups.Microvessel density inside glioma in group 2(6.51±2.71) was lower than in group 1(13.52±3.06),group 3(12.67±2.61) and control group(14.84±1.47).Conclusion:By means of inducing Th1 cytokine and suppressing Th2 cytokine and activating cytotoxic T lymphocyte,BMSCs/IL-18 induces obviously anti-tumor activity.
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The aim of this study was to investigate the effect of radiation on the expression of interleukin-6 in the bone marrow stromal cells (BMSCs) and to discuss the mechanisms by which the expression of interleukin-6 increased. The BMSCs from normal Kungming mice were cultured in vitro with the method of Dexter. The expression of interleukin-6 on the levels of protein and mRNA were detected by the uses of radio-immunity assay(RIA) and the reverse transcription polymerase chain reaction(RT-PCR) respectively. PDTC and dexamethasone, two different inhibitors of nuclear factor-kappa B, were used as controls. The result of RIA showed that the expression of interleukin-6 were distinctly increased at 1 hour, with its peak at 4 hours post irradiation in the supernatant of culture of murine BMSCs when the BMSCs were irradiated by 8.0 Gy 60 Co γ rays. The results of RT-PCR showed that the expression of IL-6 mRNA reached its peak at 8 hours. After adding NF-kappa B inhibitor in the supernatant before radiation, there was no difference between control group and radiation group in detecting the mRNA of IL-6 using RT-PCR. This study suggested that the transcription and expression of IL-6 were activated by ionizing radiation, which might be due to the reason that ionizing radiation could activate NF-kappa B.
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The study explores the expression of calcium-binding protein S100A8 and the relationship between S100A8 and cell cycle or apoptosis in bone marrow cells of mice with radiation injury. The protein/DNA bi-parameter flow cytometry method has been used to analyze the change of S100A8 protein expression and the dif-ference in cell cycle or apoptosis between S100A8 positive and S100A8 negative cells in bone marrow of mice at 6h after irradiation with 3.5 or 7 Gy 60Co γ-ray in vivo. In the normal bone marrow of mice, S100A8 positive cells are mainly granulocytes, and they are inactive in cell proliferation compared to S100A8 negative cells. At the early phase of radiation injury, the proportion of S100A8 positive cells and the expression level of S100A8 protein have no sig-nificant change in bone marrow of mice; G2/M arrest and apoptosis occur in S100A8 negative cells, but no change of cell cycle and apoptosis occur in S100A8 positive cells. These phenomena suggest that S100A8 positive cells in bone marrow of mice are radiation resistant compared to S100A8 negative cells.
S100A8
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Journal Article Effects of gamma-irradiation on the M-CSF-promoter linked to a chloramphenicol aminoacyl transferase reporter gene expressed in a clonal murine bone marrow stromal cell line Mary Ann Sakakeeny, Mary Ann Sakakeeny Department of Radiation Oncology, University of Massachusetts Medical Center, Worcester, Massachusetts, USA Search for other works by this author on: Oxford Academic Google Scholar Jean Leif, Jean Leif Department of Radiation Oncology, University of Massachusetts Medical Center, Worcester, Massachusetts, USA Search for other works by this author on: Oxford Academic Google Scholar Wilma Merrill, Wilma Merrill Department of Radiation Oncology, University of Massachusetts Medical Center, Worcester, Massachusetts, USA Search for other works by this author on: Oxford Academic Google Scholar Denise Pratt, Denise Pratt Department of Radiation Oncology, University of Massachusetts Medical Center, Worcester, Massachusetts, USA Search for other works by this author on: Oxford Academic Google Scholar Elizabeth Romanik, Elizabeth Romanik Department of Radiation Oncology, University of Massachusetts Medical Center, Worcester, Massachusetts, USA Search for other works by this author on: Oxford Academic Google Scholar Michael Mckenna, Michael Mckenna Department of Radiation Oncology, University of Massachusetts Medical Center, Worcester, Massachusetts, USA Search for other works by this author on: Oxford Academic Google Scholar T. J. Fitzgerald, T. J. Fitzgerald Department of Radiation Oncology, University of Massachusetts Medical Center, Worcester, Massachusetts, USA Search for other works by this author on: Oxford Academic Google Scholar Maureen Harrington, Maureen Harrington Indiana University, Indianapolis, Indiana, USA Search for other works by this author on: Oxford Academic Google Scholar Joel S. Greenberger Joel S. Greenberger Department of Radiation Oncology, University of Pittsburgh School of Medicine, Presbyterian University Hospital, Pittsburgh, Pennsylvania, USA Correspondence: Dr. Joel S. Greenberger, Department of Radiation Oncology, University of Pittsburgh School of Medicine, Presbyterian University Hospital, Desoto and O'Hara Streets, Pittsburgh, PA 15213-2582, USA. Search for other works by this author on: Oxford Academic Google Scholar Stem Cells, Volume 12, Issue 1, 1994, Pages 87–94, https://doi.org/10.1002/stem.5530120115 Published: 01 January 1996 Article history Received: 06 July 1993 Accepted: 31 August 1993 Revision received: 28 September 1993 Published: 01 January 1996
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Objective To investigate the radioprotective effect of genistein (GEN) on murine bone marrow stromal cells (BMSCs). Methods Adult male mice were administered orally with GEN at 24 h prior to 6.0 Gy gamma irradiation. The number of CFU-F, the adherent capability of bone marrow cells (BMCs) to BMSCs adherence layer, and the expressions of cell adhesion molecules including vascular cell adhesion molecule-1 (VCAM-1), fibronectin (Fn), and laminin (Ln) on BMSCs cultured in vitro were detected, respectively. Results The number of CFU-F, the adherent capability, the expressions of VCAM-1, Fn, and Ln on BMSCs cultured in vitro in GEN-pretreated mice were higher and recovered more rapidly than those in mice without GEN-pretreatment. Light microscopy revealed that there was no significant difference in morphology of BMSCs in both groups after irradiation injury. Conclusion These observations suggest that radioprotective properties associated with GEN are largely a consequence of the reduction of damage, the induction of cell proliferation, and the promotion of adherent capability recovery in recipient mice. These changes may ultimately ameliorate bone marrow microenvironment and therefore accelerate the rebuilding of hematopoietic system after irradiation injury.
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