结缔组织生长因子在血管紧张素Ⅱ诱导的人近端肾小管上皮细胞肥大中的作用  

Role of connective tissue growth factor in angiotensin Ⅱ induced human proximal tubular cell hypertrophy

作　　者：刘必成[1] 孙静[1] 刘宏[1] 陈珑[1] 陈琪[2] 范乐明[2] 

机构地区：[1]东南大学肾脏病研究所东南大学附属中大医院肾内科,南京210009 [2]南京医科大学病理生理学系

出　　处：《中华医学杂志》2004年第17期1470-1474,共5页National Medical Journal of China

基　　金：江苏省自然科学基金资助项目 (BK2 0 0 2 0 5 2 );江苏省医学重点人才培养基金资助项目(RC2 0 0 2 0 72 )

摘　　要：目的　探讨结缔组织生长因子 (CTGF)在介导血管紧张素Ⅱ (AngⅡ )诱导的人肾近端小管细胞肥大中的作用。方法　用 [3 H] 亮氨酸掺入、考马斯亮蓝蛋白质定量技术分别观察抗CTGF抗体对AngⅡ诱导的人肾近端小管上皮细胞株 (HK 2 )细胞内蛋白质从头合成及细胞内蛋白质含量的影响 ;用流式细胞分析技术观察抗CTGF抗体对AngⅡ诱导的细胞周期分布改变的影响 ;用扫描电镜技术观察抗CTGF抗体对AngⅡ诱导细胞形态变化的影响。 结果　AngⅡ可以显著诱导细胞内[3 H] 亮氨酸掺入和细胞内总蛋白质含量增加的作用 ,呈浓度和时间依赖性 ;抗CTGF抗体对AngⅡ诱导的细胞内总蛋白质含量、[3 H] 亮氨酸掺入量的增加有抑制作用 ,呈浓度和时间依赖性。AngⅡ作用于细胞 4 8h后 ,细胞平均直径、G0 G1期细胞百分比显著增加 (均P <0 0 1) ,抗CTGF抗体可显著抑制AngⅡ诱导的细胞直径增加和细胞周期阻滞 (均P <0 0 5或P <0 0 1)。结论　AngⅡ可以诱导肾小管上皮细胞发生肥大 ,CTGF可能介导了AngⅡ诱导的肾小管上皮细胞肥大效应。Objective Renal cell hypertrophy is an important compensatory mechanism of chronic renal diseases, which has been shown closely correlated with the activation of intrarenal renin angiotensin system. However, the exact mechanism is still uncertain. The present study was to investigate the role of connective tissue growth factor (CTGF) in mediating the effect of angiotensin (AngⅡ) induced human proximal tubular cell (HK-2) hypertrophy. Methods The cell line, HK-2, was grown in Dulbeccos′s Modified Eagle′s Medium (DMEM) containing 10% heat inactivated fetal calf serum (FCS). After rested in serum-free medium for 24 hours, The influence of anti-CTGF antibody on AngⅡ induced cell protein de novo synthesis and total protein content were determined by [3H]-leucine incorporation and Coomassie brilliant blue G250 technique respectively. Fluorescence-activated cell sorter (FACS) flow cytometer was used to analyze the effect of anti-CTGF antibody on cell cycle distribution. The change of cellular size was determined by scanning electronic microscope (SEM). Results AngⅡ significantly induced the increase of [3H]leucine incorporation in dose [AngⅡ (mol/L): 0: 6926±1034; 10 -9: 8455±2137; 10 -7: 10 741±802; 10 -5:12 945±1377] and time [AngⅡ (10 -7mol/L): 0 h: 5584±1016; 24 h: 7379±957; 48 h: 10 741±802; 72 h: 16 606±1177] dependent manner. Meanwhile, the influence of AngⅡ on cell total protein content showed the similar manner. Anti-CTGF antibody significantly inhibited the Ang Ⅱ induced above effects dose and time dependently. 48 h after the stimulation by Ang II (10 -7mol/L), the percentage of cells in G0-G1 phase (76.09%±1.82%) and the average cell diameter (20.6 μm± 3.8 μm) was significantly increased compared to the control (62.1%±2.5%, 11.9 μm±1.6 μm, P<0.01 respectively), which could markedly reversed by treatment with anti-CTGF antibody (71.68%±1.78%; 16.4 μm±3.2 μm, P<0.05, 0.01 respectively vs AngⅡ group). Conclusion Ang Ⅱ could induce the deve

关 键 词：结缔组织生长因子 血管紧张素Ⅱ 近端肾小管 上皮细胞肥大 CTGF 

分 类 号：R587.2[医药卫生—内分泌]