结缔组织生长因子介导血管紧张素Ⅱ诱导的人肾近端小管细胞肥大机制  

Mechanism of connective tissue growth factor in mediating angiotensinⅡ induced hypertrophy of renal proximal tubular cells

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

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

出　　处：《中华肾脏病杂志》2004年第3期190-193,共4页Chinese Journal of Nephrology

基　　金：江苏省自然科学基金(BK2002052);江苏省医学重点人才基金(RC2002072)

摘　　要：目的探讨结缔组织生长因子(CTGF)介导血管紧张素(Ang)Ⅱ致肾小管上皮细胞肥大的可能作用机制。方法用AngⅡ刺激体外培养的人肾近端小管上皮细胞株(HK-2),以流式细胞仪观察抗CTGF抗体(0.5μg/ml)对AngⅡ(10-7mol/L)诱导的细胞周期分布改变的影响。用RT-PCR观察不同浓度AngⅡ(0、10-9、10-7、10-5mol/L)刺激细胞48h,和AngⅡ(10-7mol/L)在不同时间点(0、24、48、72h)时HK-2细胞p27kip1mRNA表达情况;同时观察抗CTGF抗体(0.5μg/ml)对AngⅡ(10-7mol/L)诱导p27kip1mRNA表达变化的影响。用免疫细胞化学法观察抗CTGF抗体(0.5μg/ml)对AngⅡ(10-7mol/L)诱导的细胞p27kip1蛋白表达的影响。结果AngⅡ使G0～G1期细胞百分比明显增加(P<0.01),抗CTGF抗体可显著抑制AngⅡ诱导的细胞周期阻滞(P<0.05)。AngⅡ呈时间和浓度依赖性刺激p27kip1mRNA表达上调(P均<0.05),抗CTGF抗体可以显著抑制AngⅡ诱导的p27kip1mRNA表达的增加(P<0.05),免疫细胞化学显示AngⅡ可显著增加肾小管上皮细胞p27kip1蛋白表达,此作用可被抗CTGF抗体抑制。结论CTGF在AngⅡ诱导HK2细胞肥大中可能发挥重要的作用,其机制可能与抑制细胞表达p27kip1,并逆转细胞增殖周期阻滞有关。Objective To investigate the mediating mechanism of connective tissue growth factor (CTGF) in AngⅡ induced renal tubular cell hypertrophy.Methods The human renal proximal tubular cell line(HK 2) was grown in DMEM containing 10%heat inactivated fetal calf serum (FCS). The influence of anti CTGF antibody(0 5 μg/ml)on AngⅡ induced cell cycle change was analyzed by fluorescenceactivated cell sorter(FACS) flow cytometer. The dose and time response of p27kip1 mRNA expression to the stimulation of different concentrations(0,10-9,10-7,10-5mol/L)or different time intervals (0,24,48,72 h)of AngⅡwere observed by RT PCR. The effect of anti CTGF antibody(0 5 μg/ml)on AngⅡ(10-7mol/L) induced cell p27kip1 expression was observed by immunocytochemistry.Results Flow cytometer study showed that AngⅡarrested the cell cycle at G0～G1 phase(P< 0 01), which was significantly reversed by treatment with anti CTGF antibody(P< 0.05).The expression of p27kip1 mRNA was upregulated when stimulated by AngⅡin dose and time dependent manner, and the peak expression levels occurred at the concentration of 10-5mol/L and at the time of 48 h.Cocultured with anti CTGF antibody could attenuate significantly the increasing expression of p27kip1 mRNA induced by AngⅡ(P< 0 05). Immunocytochemistry revealed that AngⅡincreased the expression of p27kip1 protein(P< 0 01),which could be attenuated significantly by treatment with anti CTGF antibody(P< 0.05).Conclusion CTGF plays an important role in mediating AngⅡ induced renal hypertrophy, which may be related with its inhibition of the expression of p27kip1 and arrest of the cell cycle at G0～G1 phases.

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

分 类 号：R692.6[医药卫生—泌尿科学]