Jadus M R, Pai R, Horansky E, Wepsic H T, Kirschenbaum M A, Kamanna V S
Department of Veterans Affairs Medical Center, Long Beach, CA.
Biochim Biophys Acta. 1994 Nov 10;1224(2):181-8. doi: 10.1016/0167-4889(94)90189-9.
This study examined the ability of mesangial cells to synthesize colony-stimulating factors (CSF), cytoregulatory peptides associated with the differentiation and proliferation of hematopoietic cells. Conditioned media obtained from SV-40 transformed murine mesangial cells stimulated the growth of murine bone marrow progenitor cells of the myeloid series. Differential analysis of these cells showed the presence of both macrophages and granulocytes. Cellular identification of bone marrow colonies stimulated in response to mesangial cell conditioned media was examined by flow cytometric analysis and revealed the presence of F4/80 antigen positive macrophages (67%) and Gran-1 antigen positive granulocytes (21%). Neutralizing antibodies to macrophage colony-stimulating factor (M-CSF) and granulocyte-macrophage colony-stimulating factor (GM-CSF) but not antibody to interleukin-3 (IL-3), or stem cell factor (SCF) significantly inhibited the growth of the progenitor cells induced by mesangial cell conditioned media. Utilizing Northern blot analysis, murine mesangial cells expressed mRNA transcripts for M-CSF, GM-CSF, and granulocyte colony-stimulating factor (G-CSF). Further studies were performed to determine optimal incubation conditions for mesangial cell CSF gene expression. These studies revealed that both GM-CSF and G-CSF mRNA were maximally expressed at early time points (4 and 8 h of incubation), while M-CSF mRNA expression remained unchanged during the incubation of mesangial cells from 4-48 h. Incubation of mesangial cells with various concentrations of fetal bovine serum (FBS, 0.5-15%) markedly increased the mRNA expression of M-CSF, GM-CSF and G-CSF in a dose-dependent manner. These studies indicated that transformed murine mesangial cells are able to synthesize and secrete biologically active CSF that are associated with the migration and proliferation of circulating mononuclear cells in the glomerulus. Furthermore, observations regarding the role of duration of incubation and the media concentration of FBS on mesangial cell CSF mRNA expression may provide useful data to understand the optimal conditions for studies that examine the gene expression of basal or inducible CSF in mesangial cells.
本研究检测了系膜细胞合成集落刺激因子（CSF）的能力，CSF是与造血细胞分化和增殖相关的细胞调节肽。从SV - 40转化的小鼠系膜细胞获得的条件培养基刺激了髓系小鼠骨髓祖细胞的生长。对这些细胞的差异分析显示存在巨噬细胞和粒细胞。通过流式细胞术分析检测了响应系膜细胞条件培养基刺激的骨髓集落的细胞鉴定，结果显示存在F4/80抗原阳性巨噬细胞（67%）和Gran - 1抗原阳性粒细胞（21%）。抗巨噬细胞集落刺激因子（M - CSF）和粒细胞 - 巨噬细胞集落刺激因子（GM - CSF）的中和抗体，但抗白细胞介素 - 3（IL - 3）或干细胞因子（SCF）的抗体不能显著抑制系膜细胞条件培养基诱导的祖细胞生长。利用Northern印迹分析，小鼠系膜细胞表达了M - CSF、GM - CSF和粒细胞集落刺激因子（G - CSF）的mRNA转录本。进行了进一步研究以确定系膜细胞CSF基因表达的最佳孵育条件。这些研究表明，GM - CSF和G - CSF mRNA在早期时间点（孵育4和8小时）表达最高，而在系膜细胞孵育4 - 48小时期间，M - CSF mRNA表达保持不变。用不同浓度的胎牛血清（FBS，0.5 - 15%）孵育系膜细胞以剂量依赖方式显著增加了M - CSF、GM - CSF和G - CSF的mRNA表达。这些研究表明，转化的小鼠系膜细胞能够合成和分泌与肾小球中循环单核细胞迁移和增殖相关的生物活性CSF。此外，关于孵育时间和FBS培养基浓度对系膜细胞CSF mRNA表达作用的观察结果可能为理解检测系膜细胞中基础或诱导性CSF基因表达的研究的最佳条件提供有用数据。
