一种表现为皮肤胶原沉着病的小鼠慢性移植排斥模型的建立

摘要

Objective To establish a murine cGVHD model. Methods The BALB/e mice ( ♀ , female ) transplanted with B1O. D2 ( ♂ , male ) bone marrow( BM ) and spleen cell mixture were used to construct a murine cGVHD model with scleroderma. Syngeneic BM transplantation( BMT ) was conducted in control group. Back skin was depilated and harvested to analyze the generation of sclerodermatous graft-versus-host disease in allo-BMT mice group to compared with syngeneic group animals at day 14 post-transplantation by using histological, flow cytometric, real-time PCR and PCR analyses. Results H&E staining demonstrated that allo-BMT mice skin exhihited typical scleroderma characterized by the thickening skin,infiltration of mononuclear cell, decreasing hair follicle , and the disappearance of atrophic fat layer. Many donor cells were present in the skin of allo-BMT mice, but not in syngeneic bone marrow transplanted controls hy PCR analyses of Y-chromosome sequences. Flow cytometric analyses confirmed that increased cutaneous CD45＋ cells were accompanied by the thickening skin, in which CDllb＋( monocyte/macrophages ) and CD3 ＋( T cells ) cells were predominating in the dermal infiltrates.mRNA abundance for cutaneous RANTES, TGF-β1 and collagen Ⅰ was observed to be up-regulated in sc.lerodermatous mice compared to the syngeneic group animals. Conclusion The murine cGVHD model with scleroderma is established that can help further investigation.%目的 建立一种小鼠慢性移植排斥模型.方法 将6～8周龄BALB/c(雌性)小鼠经6 Gy 60Co全身照射后随机分为同基因移植组和异基因移植组.异基因组尾静脉输注未经照射的B10.D2(雄性)小鼠的骨髓细胞和脾细胞的混合悬液.移植术后通过小鼠性染色体基因组检测、组织病理学分析,以及流式细胞仪和实时定量PCR检测确定小鼠病变的发生.结果 病理学分析显示,异基因组小鼠主要表现为皮肤损伤,与同基因组相比,皮肤明显增厚,皮肤弹性降低,真皮层炎性细胞浸润,毛囊和脂肪减少或消失,大量的纤维组织增生,皮下组织出现明显结构改变;性染色体基因组分析显示,异基因组受者小鼠皮肤内有供者小鼠细胞的浸润;流式结果显示,异基因组小鼠皮肤内CD11b、CD45和CD3表达明显增强;实时定量PCR检测结果显示,异基因组小鼠皮肤RANTES、TGF-β1以及胶原蛋白Ⅰ mRNA水平明显升高.结论 与同基因组小鼠相比,异基因组小鼠表现为皮肤胶原沉着病损伤;CD11b+、CD45+和CD3+ 细胞以及趋化因子RANTES和细胞因子TGF-β1参与了这一病理过程的发生.本研究建立了一种表现为皮肤胶原沉着病的小鼠慢性移植排斥反应模型,为相关后续研究提供了有益的基础.