Vielhauer Volker, Anders Hans-Joachim, Mack Matthias, Cihak Josef, Strutz Frank, Stangassinger Manfred, Luckow Bruno, Gröne Hermann-Josef, Schlöndorff Detlef
Nephrologisches Zentrum, Medizinische Poliklinik, Ludwig-Maximilians-University, Munich, Germany.
Department of Animal Physiology, Ludwig-Maximilians-University, Munich, Germany.
J Am Soc Nephrol. 2001 Jun;12(6):1173-1187. doi: 10.1681/ASN.V1261173.
The infiltration of leukocytes plays a major role in mediating tubulointerstitial inflammation and fibrosis in chronic renal disease. CC chemokines participate in leukocyte migration and infiltration into inflamed renal tissue. Because CC chemokine-directed leukocyte migration is mediated by target cell expression of a group of CC chemokine receptors, this study examined the expression of CC chemokines and their receptors during initiation of tubulointerstitial fibrosis after unilateral ureteral obstruction in C57BL/6 mice. Obstructed kidneys developed hydronephrosis, tubular cell damage, interstitial inflammation, and fibrosis. From days 2 to 10, a progressive interstitial influx of F4/80+ macrophages and CD3+ lymphocytes occurred (macrophages, 4-fold; lymphocytes, 20-fold at day 10, compared with contralateral control kidneys). In parallel, the number of activated fibroblast-specific protein 1+ fibroblasts and interstitial collagen IV accumulation increased from days 2 to 10. The mRNA expression of CC chemokines (predominantly monocyte chemoattractant protein-1 [MCP-1]/CCL2, RANTES/CCL5) and their receptors CCR1, CCR2, CCR5 increased progressively from days 2 to 10. By in situ hybridization, a prominent interstitial mRNA expression of MCP-1 and RANTES and their receptors CCR2 and CCR5 localized to interstitial mononuclear cell infiltrates. MCP-1 and RANTES expression was also seen in tubular epithelial cells. Fluorescence-activated cell sorter analysis of single-cell suspensions from obstructed kidneys revealed a prominent expression of CCR2 and CCR5 by infiltrating macrophages, whereas most lymphocytes expressed CCR5 only. These data demonstrate an increased expression of MCP-1/CCL2 and RANTES/CCL5 at sites of tubulointerstitial damage and progressive fibrosis during unilateral ureteral obstruction that correlates with simultaneous accumulation of interstitial macrophages and T lymphocytes expressing the respective surface receptors CCR2 and CCR5. The chemokine receptor-mediated leukocyte influx into the tubulointerstitium could offer a new potential target for therapeutic intervention in progressive renal tubulointerstitial fibrosis.
白细胞浸润在介导慢性肾病的肾小管间质炎症和纤维化过程中起主要作用。CC趋化因子参与白细胞向炎症肾组织的迁移和浸润。由于CC趋化因子介导的白细胞迁移是由一组CC趋化因子受体的靶细胞表达介导的,本研究检测了C57BL/6小鼠单侧输尿管梗阻后肾小管间质纤维化起始过程中CC趋化因子及其受体的表达。梗阻肾脏出现肾积水、肾小管细胞损伤、间质炎症和纤维化。从第2天到第10天,F4/80+巨噬细胞和CD3+淋巴细胞逐渐向间质内浸润(与对侧对照肾脏相比,巨噬细胞在第10天增加了4倍;淋巴细胞增加了20倍)。同时,从第2天到第10天,活化的成纤维细胞特异性蛋白1+成纤维细胞数量和间质胶原IV积累增加。CC趋化因子(主要是单核细胞趋化蛋白-1 [MCP-1]/CCL2、调节激活正常T细胞表达和分泌因子/CCL5)及其受体CCR1、CCR2、CCR5的mRNA表达从第2天到第10天逐渐增加。通过原位杂交,MCP-1和调节激活正常T细胞表达和分泌因子及其受体CCR2和CCR5在间质中的mRNA表达显著,定位于间质单核细胞浸润处。在肾小管上皮细胞中也可见MCP-1和调节激活正常T细胞表达和分泌因子的表达。对梗阻肾脏单细胞悬液进行荧光激活细胞分选分析显示,浸润的巨噬细胞显著表达CCR2和CCR5,而大多数淋巴细胞仅表达CCR5。这些数据表明,在单侧输尿管梗阻期间,肾小管间质损伤和进行性纤维化部位的MCP-1/CCL2和调节激活正常T细胞表达和分泌因子/CCL5表达增加,这与表达相应表面受体CCR2和CCR5的间质巨噬细胞和T淋巴细胞的同时积累相关。趋化因子受体介导的白细胞流入肾小管间质可能为进行性肾小管间质纤维化的治疗干预提供一个新的潜在靶点。