巨噬细胞集落刺激因子对小鼠腹腔巨噬细胞抗氧化能力的影响

越来越多的实验结果证实巨噬细胞集落刺激因子在动脉粥样硬化发生过程中的重要作用，为了探讨巨噬细胞集落刺激因子和活性氧与动脉粥样硬化之间的相互关系，观察了巨噬细胞集落刺激因子对培养的小鼠腹腔巨噬细胞抗氧化酶活性的影响。结果发现巨噬细胞集落刺激因子能使小鼠腹腔巨噬细胞谷胱甘肽过氧化物酶活性提高３２％，超氧化物歧化酶的活性提高１２０％，并减轻叔丁基氢过氧化物促巨噬细胞源性泡沫细胞形成，以及提高巨噬细胞的生存数。此结果提示巨噬细胞集落刺激因子提高杭氧化酶活性的作用可能是其虽增加对氧化修饰低密度脂蛋白和胆固醇的摄入却能阻止泡沫细胞形成及动脉粥样硬化发生发展的原因之一。     

巨噬细胞集落刺激因子对RAW264.7细胞谷胱甘肽过氧化物酶活性及其基因表达的影响

为探讨巨噬细胞集落刺激因子抗单核巨噬细胞过氧化损伤的机理,以富含巨噬细胞集落刺激因子的Ｌ９２９细胞条件培养基作为细胞因子来源,采用酶活性测定、反转灵聚合酶链反应等技术,研究了巨噬细胞集落刺激因子对小鼠巨噬细胞系ＲＡＷ２６４．７细胞谷胱甘肽过氧化物酶活性及ｍＲＮＡ表达的影响。结果发现,巨噬细胞集落刺激因子可以提高ＲＡＷ２６４．７细胞硒谷胱革肽过氧化物酶及非硒谷胱甘肽过氧化物酶活性,并增强ＲＡＷ２６４     

巨噬细胞集落刺激因子对RAW264.7细胞谷胱甘肽过氧化物酶活性及其基因表达的影响

为探讨巨噬细胞集落刺激因子抗单核巨噬细胞过氧化损伤的机理 ,以富含巨噬细胞集落刺激因子的L92 9细胞条件培养基作为细胞因子来源 ,采用酶活性测定、反转录聚合酶链反应等技术 ,研究了巨噬细胞集落刺激因子对小鼠巨噬细胞系RAW2 6 4.7细胞谷胱甘肽过氧化物酶活性及mRNA表达的影响。结果发现 ,巨噬细胞集落刺激因子可以提高RAW2 6 4.7细胞硒谷胱甘肽过氧化物酶及非硒谷胱甘肽过氧化物酶活性 ,并增强RAW2 6 4.7细胞血浆谷胱甘肽过氧化物酶mRNA的表达 ,而对RAW2 6 4.7细胞磷脂氢过氧化物谷胱甘肽过氧化物酶的表达无诱导作用     

巨噬细胞集落刺激因子对小鼠腹腔巨噬细胞清道夫受体途径和细胞内胆固醇酯积聚的影响

为探索巨噬细胞集落刺激因子、清道夫受体、氧化型低密度脂蛋白与动脉粥样硬化的关系，观察了重组人巨噬细胞集落刺激因子对小鼠腹腔巨噬细胞清道夫受体途径的影响以及重组人巨噬细胞集落刺激因子对氧化型低密度脂蛋白所致细胞内胆固醇酯积聚的影响．结果发现重组人巨噬细胞集落刺激因子能增加培养的小鼠腹腔巨噬细胞表面的清道夫受体数目，使之对氧化型低密度脂蛋白的结合和降解呈现剂量和时间依赖性增加，并使细胞内胆固醇酯积聚增多．表明巨噬细胞集落刺激因子可通过增加清道夫受体数目使小鼠腹腔巨噬细胞对氧化型低密度脂蛋白的结合和降解增多，从而增加细胞内胆固醇酯含量，促进动脉壁内泡沫细胞形成．     

巨噬细胞集落刺激因子对小鼠腹腔巨噬细胞清道夫受体途径和细胞内胆固醇酯积聚的影响

为探索巨噬细胞集落刺激因子，清道夫受体、氧化型低密度脂蛋白与动脉粥样硬化的关系，观察了重组人巨噬细胞集落刺激因子对小鼠腹腔巨噬细胞清道夫受体途径的影响以及重组人巨细胞内胆固醇酯积聚的影响。结果发现重组人巨噬细胞集落刺激因子能增加培养小鼠的腹腔巨噬细胞表面的清道夫受体数目，使之对氧化型低密度脂蛋白的结合和降解呈现剂量和时间依赖性增加，并使细胞内胆固醇酯积聚增多。     

巨噬细胞集落刺激因子在动脉粥样硬化发生中的作用

动脉粥样硬化斑块损伤处可见巨噬细胞集落刺激因子表达增强，巨噬细胞集落刺激因子能通过调节核－巨噬细胞的一系列功能，影响胆固醇代谢，抑制高胆固醇血症家兔泡沫细胞形成及减少主动脉斑块面积。巨噬细胞集落刺激因子与氧化修饰低密度脂蛋白关系密切，二者可以相互作用，共同影响着动脉粥样硬化的进程。     

巨噬细胞集落刺激因子对RAW264.7细胞基质金属蛋白酶9分泌的影响

目的探讨巨噬细胞集落刺激因子致动脉粥样硬化作用是否与其影响基质金属蛋白酶表达和活性有关。方法应用酶谱法观察巨噬细胞集落刺激因子对体外培养的RAW264.7细胞基质金属蛋白酶9活性的影响;逆转录聚合酶链反应检测巨噬细胞集落刺激因子对体外培养的RAW264.7细胞基质金属蛋白酶9 mRNA表达的影响。结果与对照组相比,不同浓度(100、250和500μg/L)巨噬细胞集落刺激因子能增强RAW264.7细胞基质金属蛋白酶9的活性,并呈一定的剂量依赖性;同时,250μg/L巨噬细胞集落刺激因子可增强RAW264.7细胞基质金属蛋白酶9 mRNA表达。结论巨噬细胞集落刺激因子可诱导RAW264.7细胞基质金属蛋白酶9的表达,并可能通过这一机制影响动脉粥样硬化斑块的稳定性。     

巨噬细胞集落刺激因子与脑缺血

在中枢神经系统内，巨噬细胞集落刺激因子主要由星形细胞产生，发育期的大脑皮质神经元和小脑质细胞表达巨噬细胞集落刺激因子的受体，在脑缺血时，中枢神经系统的巨噬细胞集落刺激因子及其受体的表达均上调，补充外源性巨噬细胞集落刺激因子可明显减轻缺血性脑损伤，推测这种缺血保护作用是通过巨噬细胞集落刺激因子与受体介导的信号传导途径以及与其他因子的相互作用实现的。     

粒细胞—巨噬细胞集落刺激因子对人内皮细胞基质金属蛋白酶分泌及活性的影响

为探讨粒细胞—巨噬细胞集落刺激因子对人内皮细胞基质金属蛋白酶分泌及活性的影响，体外培养人脐静脉内皮细胞，分别应用蛋白印迹法和酶谱法检测基质金属蛋白酶2的含量和活性。结果发现，与对照组相比，不同浓度(5、25和50μg/L)粒细胞—巨噬细胞集落刺激因子可使人内皮细胞基质金属蛋白酶2的蛋白表达和活性增强，且随粒细胞—巨噬细胞集落刺激因子剂量增加而增强。提示粒细胞—巨噬细胞集落刺激因子可通过刺激人内皮细胞基质金属蛋白酶2蛋白的表达，并增强其活性，促进内皮细胞对细胞外基质的降解，从而易于动脉粥样硬化斑块内炎性细胞的渗入，内皮细胞的迁移及新血管的形成，从而对斑块的稳定性产生影响。     

巨噬细胞集落刺激因子对DEL细胞增殖作用的研究

目的 研究巨噬细胞集落刺激因子对动脉粥样硬化病变处伴随血管平滑肌细胞共同积聚的巨噬细胞的增殖作用及机制。方法 以能分化为巨噬细胞表型的ＤＥＬ细胞为离体细胞实验模型。将富含巨噬细胞集落刺激因子的１９２９细胞条件培养液或重组的人巨噬细胞集落刺激因子加入体外培养的处于静止期的ＤＥＬ细胞,以氚标胸腺嘧啶核苷或５－溴－悄嘧啶 核苷掺入法分别检测ＤＥＬ细胞ＤＮＡ的合成,另以Ｎｏｒｔｈｅｒｎ ｂｌｏｔ法检测ＤＥ     

Constitutive activation of the MAPK pathway mediates v-fes-induced mitogenesis in murine macrophages

Fes is a nonreceptor tyrosine kinase expressed at the highest level in macrophages. We previously showed that the overexpression of c-fes in murine macrophages of the BAC-1.2F5 cell line renders these cells independent of macrophage colony-stimulating factor (MCSF) for survival and proliferation, although no direct relationship could be established between tyrosine-phosphorylated substrates of Fes- and MCSF receptor-dependent signaling and mitogenesis. In this study, we investigated whether the mitogen-activated protein kinase (MAPK) pathway is involved in the growth factor-independent growth of v-fes-overexpressing macrophages. We found a constitutively increased phosphorylation of extracellularly regulated kinase (ERK) in v-fes-overexpressing macrophages as compared with mock-infected cells. This finding was associated with activation of mitogen/extracellular signal-regulated kinase (MEK) and with constitutive localization of ERK in the nucleus. Treatment of v-fes-overexpressing cells with the MEK-specific inhibitor PD98059 markedly reduced cell growth, hyperphosphorylation, and nuclear localization of ERK, indicating that the MAPK pathway mediates the mitogenic effect of v-fes. (Blood. 2000;95:3959-3963)     

巨噬细胞集落刺激因子与血管平滑肌细胞增殖

目的一些以血管平滑肌细胞（ＶＳＭＣ）增殖为特点的血管疾病，在病变部位常有巨噬细胞浸润。本研究巨噬细胞集落刺激因子（ＭＣＳＦ）在ＶＳＭＣ生长调节中的作用。方法实验采用培养大鼠主动脉ＶＳＭＣ，细胞增殖观察指标采用氚标胸腺嘧啶核苷掺入法，并用Ｎｏｒｔｈｅｒｎｂｌｏｔ技术测定原癌基因表达。结果（１）Ｌ９２９细胞上清液（富含ＭＣＳＦ）及重组ＭＣＳＦ以剂量依赖关系刺激氚标胸腺嘧啶核苷掺入；（２）ＶＳＭＣ在接受刺激后表达某些原癌基因，如ｃｆｏｓ、ｃｍｙｃ、ｅｒｇ１和ＪｕｎＢ；（３）凝血酶、ＰＤＧＦ、ｂＦＧＦ与ＭＣＳＦ在促增殖作用上具有协同作用。结论ＭＣＳＦ与其它生长因子协同作用，通过自分泌／旁分泌机制调控ＶＳＭＣ增殖，从而可能在血管病变的形成和进展中起重要作用     

Potential role of macrophage colony-stimulating factor in the proliferation of DEL cells.

The replication and activation of both vascular smooth muscle cells and macrophages, which have previously entered the arterial wall, are key events in the atherosclerotic process. The importance of macrophage colony-stimulating factor (MCSF) in control of the growth/proliferation of both cell types confers to this compound a central role in the development of vascular lesions. In order to gain insight into the mechanisms of macrophage proliferation, we investigated the effect of MCSF upon the proliferation of DEL cells. DEL cells constitute a monocyte/histiocytic cell line that differentiates along a macrophage lineage following exposure to phorbol ester. DEL cells constitutively express MCSF, and its receptor MCSFR is encoded by c-fms. We examined whether MCSF might play a role in the proliferation of cultured DEL cells. [3H]Thymidine or 5-bromo-2-deoxyuridine (BrdU) incorporation was measured following the addition of recombinant MCSF or L929 cell supernatant (as a source of MCSF) to quiescent DEL cells. In DEL cells, serum-free L929 cell supernatant induced DNA synthesis in a dose-dependent manner, and such an effect could be blunted by pretreatment of L929 cell supernatant with anti-mouse MCSF antibody. In these cells, DNA synthesis could also be triggered in a dose-dependent manner by the addition of recombinant human MCSF (rh MCSF) or thrombin. These findings clearly show that MCSF influences DEL cell proliferation and suggest an autocrine loop activation. They indicate that MCSF plays an important role in the development of vascular lesions, which occur during atherosclerotic progression.     

Light诱导类风湿关节炎滑膜细胞向破骨细胞转化

目的 观察Light在类风湿关节炎(RA)滑膜细胞向破骨细胞转化过程中的作用.方法 取8例RA患者滑膜组织,胶原酶消化获取滑膜细胞,每例滑膜细胞分成5份培养,第1组加入巨噬细胞集落刺激因子(MCSF)作阴性对照,第2组加入MCSF和LIGHT,第3组加入MCSF和核因子(NF)-κB受体激动剂配体(RANKL),第4组加入MCSF、LIGHT和RANKL,第5组加入LIGHT.体外培养2周后,行抗酒石酸酸性磷酸酶(TRAP)染色,F肌动蛋白(F-actin)染色以及象牙片上骨吸收陷窝观察破骨细胞的形成和活性.结果 第1组和第5组TRAP(-),F-actin(-),象牙片上无骨吸收陷窝形成;第2组TRAP(+),F-actin(+),骨陷窝形成(+),多核破骨细胞呈圆形和椭圆形,体积较小,骨陷窝分散,体积较小;第3组TRAP(++).F-actin(++),骨陷窝形成(++),多核破骨细胞体积大,骨陷窝较多,体积大,形态不规则;第4组TRAP(+++),F-actin(+++),骨陷窝形成(++++),多核破骨细胞更多,骨陷窝大且有融合趋势.结论 Light能诱导RA滑膜细胞向破骨细胞转化,并能促进RANKL诱导滑膜细胞向破骨细胞转化的能力.     

Large variations in human foam cell formation in individuals: a fully autologous in vitro assay based on the quantitative analysis of cellular neutral lipids

The transformation of monocyte-derived macrophages into lipid-laden foam cells constitutes a characteristic and crucial event in the development of the earliest atherosclerotic lesions. We investigated whether the propensity to form foam cells varies among individuals. We developed a fully autologous foam cell assay based on a recently developed novel culture technique for human monocyte-derived macrophages (Wintergerst ES, Jelk J, Asmis, R. Differential expression of CD14, CD36 and the LDL receptor on human monocyte-derived macrophages. A novel cell culture system to study macrophage differentiation and heterogeneity, Histochem. Cell Biol. 1998;110:231-241). Thin layer chromatography and laser densitometry were used to determine cholesterol, triglyceride and cholesteryl ester levels in human macrophages. Aggregated LDL obtained by vortexing was found to be a reproducible stimulus of foam cell formation in human macrophages. In our hands, Cu(2+)-oxidized LDL also induced cholesteryl ester accumulation, but only when vortexed. We found that foam cell formation in an individual varied by less than 25% over a 10-month period. In contrast, we observed a sevenfold difference in foam cell formation among eight male volunteers. The transfer of foam cells into culture medium with freshly thawed autologous serum resulted in a 75% regression within 1 week, independent of the amount of cellular cholesteryl esters accumulated. Foam cell formation correlated neither to serum nor to cellular cholesterol and triglyceride levels. The propensity to form foam cells could therefore represent a novel indicator of individual risk of atherogenesis.     

Breast tumor cell TACE-shed MCSF promotes pro-angiogenic macrophages through NF-κB signaling

Most deaths associated with breast cancer, the most common malignancy in women, are caused by metastasis. Tumor associated macrophages significantly contribute to breast cancer progression and development of metastasis through the promotion of angiogenesis which involves a central regulator of macrophage functions: nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB). Macrophages are activated by macrophage colony stimulating factor (MCSF) and chemokine (C–C motif) ligand 2 (CCL2) to secrete angiogenic factors including vascular endothelial growth factor (VEGF). The release of MCSF from tumor cells is mediated by ectodomain shedding through tumor necrosis factor alpha converting enzyme activation (TACE). Here we determined whether tumor cells TACE-shed MCSF promotes angiogenesis through activation of the NF-κB pathway in macrophages and the subsequent release of VEGF. These interactions were modeled in vitro using a panel of mammary cells mimicking the breast cancer progression from normal murine mammary gland cells to metastatic 4T1 cells along with J774 macrophages, all derived from BALB/c mice. TACE and MCSF expressions were higher in metastatic cells compared to epithelial cells (p < 0.05). Tumor conditioned medias activated the expression of VEGF by macrophages through stimulation of the NF-κB pathway and resulting macrophage secretions that promoted high levels of endothelial cell tubes. Furthermore, the combinations of CCL2, also highly expressed by tumor cells, and MCSF promoted pro-angiogenic macrophages. These results highlight the key role of tumor cell TACE-shed MCSF and secreted CCL2 in stimulating pro-angiogenic macrophages.