| PrefaceAs one of the major complication after allogenic bone marrow transplantation (BMT) , acute graft - versus - host disease ( aGVHD) has a bad influence on the survival rate of patients. , CsA-at present one of the regular immunosuppressive agent for inhabiting aGVHD in clinic - has serious side - effect and toxic effect, especially on liver and kidney; at the same time it can lead to severe infection as well as tumor. Although Monoclonal antibody can inhibit immunologic rejection reaction , it can lead to the repea of Leukemia because it eliminate graft - versus - Leukemia ( GVL) effect. Therefore it is an urgent problem to seek a new effective method to prevent aGVHD, which is mild and have little side - effect. We have proved that hyper-baric oxygen (HBO) could reduce the quantity of T cell and its subsets, inhibit the production of IL -2 and the expression of IL -2R, repress the reaction of T cell to IL - 2 and restrain the expression of LFA -1 on the surface of T cell. In addition, HBO can prolong the survival time of transplanted skin pieces, and improve the microcircu-latory oxygen to injured tissue, and anti - infection, so it has remarkably advantage in inhabiting aGVHD. So the paper observed the effect of HBO and on the mRNA of IL-2, TNF-a,IL-4,IL-10 in the spleen cell after allogenic bone marrow transplantation, we probe the mechanism of HBO antagonizing aGVHD and provided experimental data to clinical application of HBO.MethodDuplication of the model of aGVHDDonor mice BALB/C, female, receptor mice C57BL/6 male, ten to twelve weeks, 20 - 22g ( Laboratory Animal Center Of China Medical University). The recipient mice were exposed to 1800cGY total body radiation (TBI) delivered by a linear accelerator at a dose rate of 200cGY /min within 24 hours before BMT. The single cell suspensions suspended in RPMI1640 were obtained from the spleens and bone marrow of the donor mice sterilely at 0 ℃ All recipient mice were given 1 × 10 bone marrow cells and 5 × 106 spleen cells.Treatment of animalsAfter radiation the recipient mice were divided into four groups randomly.A; syngenie bone marrow transplantation group, the recipient mice received no further therapy;B: allogeneic bone marrow transplantation group, the recipient mice received no further therapy;C: CsA + MTX group, the recipient mice were injected into CsA and MTX;D: donor mice HBO group, the recipient mice were applied to HBO;E: donor and recipient mice HBO group, both donor and recipient mice were treated with HBO.The donor mice of D and E group were disposed by 99.9% oxy-gen with 0.25MPa for 1.5 hour once a day on day - 3, - 2, - 1 before BMT. From 1 day to 10 day post BMT, the recipient mice of C group were injected CsA (2. 5mg/kg? day) intraperitionally, the recipient mice of E group were treated with HBO as described above, and on day 1,3 ,5,9 post BMT the recipient mice of C group were injected MTX (7mg/m2) simultaneously. After BMT all the recipient mice were bred in sterile room.Assay of cytokine mRNAPut the recipient mice to death on day 11 after BMT, take its spleen tissue , extract all RNA from spleen cells and detect the change of cytokine IL-2, TNF-a,IL-4, IL-10mRNA expression by method of RT - PCRStatistical analysis: values are mean + SEM. T - test was used except for the survival rate data.RESULTRecipient mice were monitored for clinical signs of aGVHD following BMT. The appearance of allogeneic bone marrow transplantation group was characteristic of aGVHD including hunched posture, poor appetite and patchy alopecia. In contrast CsA + MTX group and HBO groups seldom exhibited the sign of aGVHD. But the CsA + MTX group was not as active as the two HBO groups and showed diarrhea because of the side effects of immunosuppressive agent.Survival of recipient mice was followed for 11 days post BMT. The survival rate of two HBO groups ( D:9/11;E:9/10) and syngeniegroup( A:9/10) was much higher than allogeneic bone marrow transplantation group ( B: 8/17 ) an... |
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