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目的:进一步探讨结缔组织生长因子(connectivetissuegrowthfactor,CTGF)反义寡核苷酸对血管紧张素Ⅱ(AngⅡ)诱导的肾小管细胞肥大的影响。方法:采用体外培养的人肾近端小管上皮细胞株(HK2),分别采用考马斯亮蓝法、流式细胞分析技术及扫描电镜和透射电镜,观察CTGF反义寡核苷酸对AngⅡ诱导的细胞内总蛋白含量、细胞周期分布改变及细胞直径和超微结构改变的影响。结果:AngⅡ干预HK2细胞48h后,细胞内总蛋白含量显著增加(P<0·01);这种作用可被CTGF反义寡核苷酸显著抑制,且呈时间和浓度依赖性。AngⅡ干预HK2细胞后,细胞大部分阻滞在G0-G1期(P<0·01),而CTGF反义寡核苷酸可逆转这种周期阻滞现象(P<0·05)。经AngⅡ干预的HK2细胞,细胞平均直径显著增加,细胞超微结构显示表面微绒毛减少、细胞内粗面内质网增多、线粒体减少、高尔基体增加,这些作用均可被CTGF反义寡核苷酸所抑制。结论:CTGF反义寡核苷酸对AngⅡ诱导HK2细胞肥大具有显著的抑制作用,提示CTGF可能介导了AngⅡ诱导的肾小管上皮细胞肥大。
OBJECTIVE: To further investigate the effects of connective tissue growth factor (CTGF) antisense oligonucleotide on angiotensin Ⅱ (AngⅡ) -induced renal tubular cell hypertrophy. Methods: Human renal proximal tubule epithelial cell line (HK2) was cultured in vitro. Coomassie brilliant blue staining, flow cytometry, scanning electron microscopy and transmission electron microscopy were used to observe the effect of CTGF antisense oligonucleotide on AngⅡ-induced cells Total protein content, cell cycle distribution and cell diameter and ultrastructure changes. RESULTS: After treated with AngⅡ for 48 h, the total cellular protein level was significantly increased (P <0.01). This effect was significantly inhibited by CTGF antisense oligonucleotide in a time-and concentration-dependent manner. After the intervention of HKⅡ, most of the cells were arrested in G0-G1 phase (P <0.01), while CTGF antisense oligonucleotide reversed this cycle arrest (P <0.05). The average diameter of HK2 cells treated with Ang Ⅱ was significantly increased, the ultrastructure of the cells showed reduced microvilli, increased endoplasmic reticulum, reduced mitochondria and increased Golgi, all of which were inhibited by CTGF antisense oligonucleotide Acid inhibited. CONCLUSION: CTGF antisense oligonucleotide significantly inhibits the hypertrophy of HK2 cells induced by AngⅡ, suggesting that CTGF may mediate angiotensin Ⅱ induced renal tubular epithelial cell hypertrophy.