昆明山海棠提取物对异基因骨髓移植小鼠调节性T细胞的影响

作者	单位
李晟	中国人民解放军空军航空医学研究所附属医院内二科
徐开林	徐州医学院附属医院血液科
李振宇	徐州医学院附属医院血液科
潘秀英	徐州医学院附属医院血液科

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中文摘要:

目的研究昆明山海棠(THH)提取物减轻异基因骨髓移植小鼠移植物抗宿主病(GVHD)作用及可能机制。方法BALB/c受鼠(H-2d)清髓性预处理后输注C57BL/6供鼠(H-2b)骨髓细胞和脾细胞混合液,随机分4组:对照组、环孢素A(CsA)组、THH组、THH+CsA联药组(联药组)。观察移植后受鼠GVHD发生情况和嵌合体植入情况,流式细胞术测定受鼠外周血CD4+CD2+5T细胞百分率以及实时荧光定量PCR方法检测受鼠脾细胞内Foxp3mRNA表达水平。结果(1)10天时CsA组、THH组、联药组CD4+CD2+5T细胞数量开始增加,15天达高峰,之后开始下降,30天时联药组仍维持较高水平;(2)CsA组、THH组、联药组受鼠脾细胞内Foxp3mRNA表达水平在15天达高峰,联药组最高,之后逐渐下降,联药组下降最缓慢,30天时仍维持较高水平;(3)CsA组、THH组、联药组受鼠生存时间均长于对照组(P<0.05);(4)CsA组、THH组、联药组GVHD临床表现和病理改变均比对照组轻;(5)对照组、CsA组、THH组、联药组受鼠骨髓细胞表面H-2b百分率均>95%。结论THH可减轻异基因骨髓移植小鼠GVHD,延长其生存时间,可能与升高CD4+CD2+5T细胞水平及促进Foxp3mRNA表达而发挥免疫抑制效应有关。THH和CsA小剂量联合应用有协同作用。

英文摘要:

Objective To study the effect of Tripterygium hypoglaucum(level)Hutch(THH)extraction in alleviating the graft-versus-host disease(GVHD)in mice after allogeneic bone marrow transplantation and to explore its mechanism.Methods Mixture of 2×107 bone marrow cells and 2×107 spleen cells obtained from C57BL/6 mouse were transplanted into the myelo-ablative irradiated inbred BALB/c mouse(H-2d).The experimental mice were designed into 4 groups:the control group(A),the Cyclosporine A(CsA)group(B),the THH group(C),and the CsA + THH group(D).Condition of GVHD and transplanted chimera were observed,percentage of CD+4CD+25T cell in peripheral blood were detected by flow cytometry,and the expression of Foxp3 mRNA in splenocyte at different time points were determined by real-time FQ-PCR.Results CD+4CD+25T cell in Group B,C and D increased from the 10th day,peaked on the 15th day,descended after then,but the level in Group D maintained higher than that in the other three groups(P<0.05).Expression of Foxp3 mRNA in recipients’ splenocytes increased gradually,reached peak on the 15th day and followed with gradual decreasing,the highest peak and the slowest decreasing was shown in Group D,so the level was still rather higher in Group D on the 30th day.The survival time in the three treated group(B,C and D)were all longer than that in the control group A(P<0.05).GVHD signs and pathologic changes in the three treated groups were milder than in the control group.Allogeneic chimerism rates(H-2d)in all groups were over 95%.Conclusion THH could prevent GVHD in mice after allogeneic bone marrow transplantation,and prolong their survival time,the mechanism is possibly related with its immunosuppressive effect in raising CD+4CD+25T cells and promoting the Foxp3 mRNA expression.THH combining with small dose CsA shows synergic effect in preventing GVHD.

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