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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 Gy60Co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ⅠmRNA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。
Objective To establish a chronic rejection model in mice. Methods BALB / c (female) mice of 6-8 weeks old were randomly divided into two groups: the same gene transplantation group and the allogeneic transplantation group. Allogeneic tail vein was infused with a mixed suspension of bone marrow cells and splenocytes from non-irradiated B10.D2 (male) mice. After transplantation, genomic detection of mouse sex chromosomes, histopathological analysis, and flow cytometry and real-time quantitative PCR were used to determine the occurrence of the lesion in mice. Results Pathological analysis showed that allogeneic mice mainly showed skin lesions. Compared with the same genomic group, the skin thickens, the skin elasticity decreases, the infiltration of inflammatory cells in the dermis, hair follicles and fat decrease or disappear, and a large number of fibrous tissue hyperplasia , Subcutaneous tissue showed obvious structural changes; genomic analysis showed that allogeneic recipient mouse skin infiltration of donor cells; flow cytometry showed that allogeneic mice skin CD11b, CD45 and CD3 expression was significantly enhanced Real-time quantitative PCR results showed that the levels of RANTES, TGF-β1 and collagen Ⅰ mRNA in the skin of allogeneic mice were significantly increased. CONCLUSIONS: Allogeneic mice showed collagen-induced skin lesions compared with those of the same genomic mice. CD11b +, CD45 + and CD3 + cells as well as chemokine RANTES and cytokine TGF-β1 were involved in this pathological process. In this study, we established a model of chronic allograft rejection in mice with dermal collagen deposition, which provides a useful basis for subsequent studies.