摘要
目的 :观察外周血单核巨噬细胞 (MO/MAC)条件培养基 (M -CM)对肾小管上皮细胞 (RTEC)直接作用的生物学效应并探讨可能的作用机制。方法 :应用正常人外周血M -CM刺激人近端肾小管上皮细胞 (HK - 2 ) ,以[3 H]-TdR掺入法检测HK - 2细胞DNA合成、Westernblot法检测骨桥蛋白 (OPN)和α -平滑肌肌动蛋白 (α -SMA)表达、间接抑制ELISA法检测纤连蛋白 (FN)分泌。进一步采用白细胞介素 - 10 (IL - 10 )和转化生长因子 - β1(TGF - β1)中和抗体进行拮抗。结果 :①M -CM可促进HK - 2细胞DNA合成、α -SMA表达及FN分泌。②TGF - β1中和抗体(5mg/L)与M -CM同时作用于HK - 2细胞 ,其α -SMA表达和FN分泌均明显低于M -CM单独作用组。③IL - 10(2 0 μg/L)与M -CM同时作用组的HK - 2细胞 ,α -SMA表达和FN分泌亦明显低于M -CM组 ;IL - 10预孵育MO/MAC组 ,HK - 2细胞α -SMA表达也明显低于M -CM组。结论 :单核巨噬细胞可直接诱导肾小管上皮细胞增殖、表型转化以及分泌细胞外基质增加 ;其分泌的TGF - β1及炎性细胞因子可能参与介导上述效应。
AIM: To observe the direct effects of peripheral blood monocytes/macrophages (MO/MAC) on renal tubular epithelial cells (RTEC),and further probe into the possible mechanisms. METHODS: Conditioned medium(M-CM) of human peripheral blood MO/MAC was collected and added to HK-2,a human renal proximal tubular cell line.After incubation with M-CM for 24 hours,HK-2 cells were detected for DNA synthesis by -TdR incorporation,osteopontin (OPN) and α-smooth muscle actin (α-SMA) expression by Western blot,and fibronectin(FN) secretion by ELISA.Furthermore,anti-TGFβ_1 neutralizing antibody and interlukin-10(IL-10) were used separately to antagonize the effects of M-CM on HK-2 cells. RESULTS: ①DNA synthesis,α-SMA expression and FN secretion were all increased in HK-2 cells when incubated with M-CM.②When adding with anti-TGFβ_1 neutralizing antibody (5 mg/L) in the M-CM,the degree of upregulation of α-SMA and FN in HK-2 cells was much lower than that stimulated by M-CM alone.③M-CM added with IL-10 (20 μg/L) had a weaker ability to induce the increasing in α-SMA expression and FN excretion in HK-2 cells, compared with M-CM itself alone.M-CM from MO/MAC preincubated with IL-10 caused a lower upregulation of α-SMA expression in HK-2 cells than M-CM from non-preincubated MO/MAC. CONCLUSION: MO/MAC can directly induce proliferation,transdifferentiation and extracellular matrix secretion in RTEC.TGFβ_1 and proinflammatory cytokines secreted by MO/MAC might be involved in the aboveeffects.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2002年第10期1217-1221,共5页
Chinese Journal of Pathophysiology
基金
国家自然科学基金资助项目 (No .39770 346 )
教育部跨世纪人才基金 (39910 2 10 4 74 - 2 31-C0 4 )资助项目
关键词
巨噬细胞
肾小管
上皮细胞
印迹法
蛋白质
酶联免疫吸附测定
Macrophages
Kidney tubules
Epithelial cells
Blotting,western
Enzyme-linked immunosorbent assay
