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目的探究慢性移植排斥反应诱导小鼠肾炎的发生机制,为下一步药物治疗提供实验依据。方法建立DBA/2→B6D2F1(DBA/2J×C57BL/6)移植小鼠模型,通过尿蛋白检测以及组织病理学分析确定小鼠肾炎的发生;利用直接免疫荧光标记技术以及实时定量PCR分析慢性移植排斥反应诱导小鼠肾炎发生过程中T、B淋巴细胞的活化情况,以及靶组织器官中各种因子或蛋白的变化。结果与同系基因型移植对照组(B6D2F1→B6D2F1)小鼠相比,异体移植实验组(DBA/2→B6D2F1)小鼠脾细胞中CD4+CD69+、CD8+CD69+双阳性细胞百分比均明显增加,B220+I-Ab+双阳性细胞的I-Ab表达显著增强,提示T、B淋巴细胞的活化;另外,在实验组小鼠肾组织中,趋化因子如单核细胞趋化蛋白-1(MCP-1)、受激活调节正常T细胞表达和分泌因子(RANTES)、B淋巴细胞趋化因子(BLC)以及淋巴细胞趋化因子(Ltn)表达上升;炎性细胞因子(如IL-6、IL-1β)表达显著增强;促纤维生长因子如转化生长因子β1(TGF-β1)、结缔组织生长因子(CTGF)以及纤维增生相关蛋白如Ⅰ型胶原蛋白(collagenⅠ)的mRNA表达水平明显上升,提示组织炎症以及纤维化的发生。结论慢性移植排斥反应诱导肾炎发生可能与激活淋巴细胞以及炎症因子、趋化因子和促纤维生长因子的分泌表达有关。
Objective To investigate the mechanism of chronic kidney allograft rejection induced by nephritis in mice and provide experimental evidence for further drug treatment. Methods The mouse model of DBA / 2 → B6D2F1 (DBA / 2J × C57BL / 6) transplantation was established. The incidence of nephritis in mice was determined by urinary protein and histopathological analysis. Direct immunofluorescence labeling and real- Exclusion reaction induced the activation of T and B lymphocytes in mouse nephritis and the changes of various factors or proteins in target tissues and organs. Results Compared with the B6D2F1 → B6D2F1 mice, the percentages of CD4 + CD69 +, CD8 + CD69 + double positive cells in the spleen cells of allogeneic transplantation mice (DBA / 2 → B6D2F1) + I-Ab + double positive cells significantly increased, suggesting T, B lymphocyte activation; In addition, in the experimental group of mouse kidney tissue, chemokines such as monocyte chemoattractant protein-1 (MCP- 1), upregulated expression of RANTES, B lymphocyte chemotactic factor (BLC) and lymphocyte chemotactic factor (Ltn) in activated normal cells. Inflammatory cytokines such as IL-6, IL- 1β) was significantly increased. The mRNA expression of fibrogenic growth factors such as transforming growth factor β1 (TGF-β1), connective tissue growth factor (CTGF) and fibrinolytic related protein (collagen Ⅰ) Inflammation and the occurrence of fibrosis. Conclusion Chronic rejection induced nephritis may be related to the activation of lymphocytes and the secretion of inflammatory cytokines, chemokines and fibroblast growth factor.