Regulation of macrophage functions by cytokine and 1,25-dihydroxyvtamin D_3 on glomerulonephritis

新月体肾小球肾炎是发生在肾小球毛细血管外并累及足细胞的许多严重损害的一个代表。新月体形成细胞是来自肾小球上皮细胞还是单核/巨噬细胞，这一问题一直备受争议，目前还没有得到澄清。为探讨新月体肾小球肾炎中足细胞的病理生理机制，我们试图建立足细胞的培养方法，重点研究足细胞与巨噬细胞之间的关系。从正常大鼠肾小球分离培养出的未知来源的非贴壁细胞数量增加，第11天达到3.5x105/ml。它们表现出巨噬细胞的几个特征，特异性抗原和酶的表达，形态和H_2O_2的产生。它们表达FxlA，但缺乏Thy1.1或因子VIII的表达。培养3-11天的形态学研究显示…肾小球基底膜(GBM)上有足突的细胞转变为巨噬细胞(MC)，并从肾小球迁移。免疫荧光双重染色显示，第8天从肾小球表面迁移的细胞均为抗Podocalyin和ED-1阳性。免疫电子显微镜显示ED-1阳性细胞定位于肾小球系膜上，抗巨噬细胞抗体和-Thy1.1抗体与补体共同作用不能减少MC的数量，而氨基核苷嘌呤霉素可显著减少MC的数量。经γ射线照射后肾小球巨噬细胞数量的显著减少并不能导致骨髓嵌合大鼠肾小球培养的MC表达Ia抗原，表明MC不是来源于骨髓细胞。巨噬细胞集落刺激因子促进肾小球系膜细胞向MC转化，粒细胞-巨噬细胞集落刺激因子在第8天随细胞数增加而明显增强。结论：正常大鼠肾小球培养中足细胞向MC转化，CSFS可促进这种转化。这些结果为新月形成细胞的起源提供了一个全新的见解。较少

One representative of a number of severe lesions that occur outside the glomerular capillaries and involve podocytes is crescentic glomerulonephritis. The question of whether the crescent-forming cells are derived from glomerular epithelial cells or monocytes/macrophages is highly controversial and has not yet been clarified. To investigate pathophysiology of podocyte in crescentic glomerulonephritis, we attempted to establish methods for culturing cells confirmed to be derived from podocytes, focusing particularly on the relationship between podocytes and macrophages.Non-adherent cells of unknown origin that grew from normal rat isolated glomerular cultures increased in number, reaching a total of 3.5 x 10^5/ml on day 11. They showed several characteristics of macrophages, the expression of specific antigens and enzyme, morphology, and production of H_2O_2. They expressed FxlA,but lacked the expressions of Thy1.1 or factor VIII.A morphological kinetic study on day 3-11 of culture show … More ed that the cells with foot processes on the glomerular basement membrane (GBM) changed into macrophagic cells (MC) and migrated from the glomeruli. Immunofluorescence double staining indicated that the cells migrated from glomerular surface on day 8 were both anti-podocalyxin and ED-1 positive. Furthermore, immunoelectron microscopy revealed that the ED-1 positive cells were located on GBM.Pretreatment with an anti-macrophages and -Thy1.1 antibodies both with complement did not reduce the number of MC,while pretreatment with puromycin aminonucleoside predominantly reduced the number of MC.Predominant decrease in the number of glomerular macrophage by gamma-irradiation did not result in the reduction in the number of MC.MC derived from glomerular cultures of bone marrow chimeric rats expressed the Ia antigen originated from recipient, indicating that MC is not derived from bone marrow cells. Macrophage colony-stimulating factor accelerated the speed of the change into MC and granulocyte-macrophage colony-stimulating factor dramatically enhanced its degree with increase of cell number on day 8.We concluded that podocytes change into MC in normal rat glomerular culture and the change is enhanced by CSFs. The results provide a completely new insight into the origin of crescent forming cells. Less