CCR7/CCL21途径在循环成纤维细胞识别与沉积导致移植肾纤维化的作用及机制研究

项目名称： CCR7/CCL21途径在循环成纤维细胞识别与沉积导致移植肾纤维化的作用及机制研究

项目编号： No.81470975

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 医药、卫生

项目作者： 周洪澜

作者单位： 吉林大学

项目金额： 66万元

中文摘要： 移植肾间质纤维化是导致慢性移植肾失功、影响移植肾长期存活的最主要原因。虽然已知纤维细胞与纤维化关系密切，但其来源及作用途径尚未阐明。近年文献报道，骨髓来源的循环成纤维细胞可导致器官纤维化，但移植肾间质纤维化过程中，循环成纤维细胞是否参与、如何进行肾脏识别与沉积并不清楚。我们前期工作显示CCL21/CCR7通路对移植肾间质纤维化成纤维细胞的迁移有重要作用。由此，我们假设CCR7/CCL21途径可调控循环成纤维细胞在移植肾中的识别与沉积，并导致间质纤维化。为验证假设，我们拟利用雌性Fisher344大鼠作为供体，雄性Lewis大鼠作为受体构建移植肾间质纤维化的模型，通过Y染色体方法检测成纤维细胞的来源；采用成纤维细胞培养，CCR7siRNA靶向转染等方法，观察CCL21/CCR7通路在循环成纤维细胞肾脏识别与沉积中的作用及机制，为延缓移植肾纤维化的发生、发展及防治慢性移植肾失功提供方法。

中文关键词： 肾脏移植；慢性移植肾肾病；肾间质纤维化；成纤维细胞；CCR7/CCL21途径

英文摘要： Despite current improvements in the longevity of renal allografts, the transplanted kidney typically loses its function before the patient dies. Although a variety of factors contribute to the renal graft loss, chronic allograft nephropathy (CAN) is recognized as the leading cause of renal allograft failure after the first year following transplantation.Histological manifestations of CAN include interstitial fibrosis, tubular atrophy and dilation, accumulation of fibroblasts, glomerulosclerosis, and chronic inflammatory cell infiltration.Fibrosis is the predominant pathological change of chronic allograft kidney.Nevertheless, the precise pathogenesis mechanisms of renal fibrosis remain poorly understood. Since fibroblasts play a critical role in the progression of fibrosis, expression of fibroblast surface protein (FSP), which correlates with the number of fibroblasts, may serve as a biomarker for evaluating fibrosis of kidney allografts. A number of growth factors and cytokines have been implicated in the development of renal fibrosis,Our results indicate that FSP, CCL21, and CCR7 are localized in the interstitial fibroblasts and renal tubular epithelium cells, suggesting the involvement of the CCL21/CCR7 pathway in the progression of renal allograft fibrosis. Hence, these observations provide evidence for the molecular and cellular mechanisms of renal allograft fibrosis, and may help to develop a therapeutic target for allograft failure following kidney transplantation.Chemokines and chemokine receptors are involved in the progression of acute and chronic renal disease . Expression of chemokine receptors, such as CCR7, CCR2 and CXCR4, have been detected in fibrocytes isolated from humans and mice. One of the chemokine receptors, CC-chemokine receptor 7 (CCR7), and its ligand chemokine CC-ligand 21 (CCL21, also known as the secondary lymphoid tissue chemokine) have been shown to regulate trafficking of fibrocytes into the kidney, which leads to renal fibrosis. However, the potential involvement of the CCL21/CCR7 pathway in fibrosis of kidney allografts following transplantation remains undetermined. In the present study, we investigate the distribution and expression of FSP, chemokine CCL21 and chemokine receptor CCR7 in renal allograft biopsies obtained from patients following kidney transplantation. In our study, the model are constructed by using female Fisher344 rat as donator and male Lewis rat as receptor. And the origin of fibroblasts are detected by Y chromatin body detection. Fibroblasts culture，flow cytometry sorting，CCR7siRNA transfection, Fibroblasts intravenous injection and intravenous blocking CCL21/CCR7 pathway are used to study the role of circulating fibrocytes and CCL21/CCR7 signal in interstitial formation of transplant kidney

英文关键词： transplan kidney;chronic allograft nephropathy;interstitial fibrosis;fibrocytes;CCR7/CCL21 pathway