结缔组织生长因子介导转化生长因子促肾小管上皮细胞转分化  

Connective tissue growth factor mediates TGF-p to regulate tubular epithelial-myofibroblast transdifferentiation in vitro

作　　者：张海燕[1] 李幼姬[2] 叶任高[2] 李晓燕[2] 杜勇[2] 孔庆瑜[2] 梁鸣[2] 余学清[2] 

机构地区：[1]广东省佛山市第一人民医院肾内科 [2]中山大学附属第一医院肾内科,广州510080

出　　处：《中华肾脏病杂志》2003年第6期360-364,共5页Chinese Journal of Nephrology

基　　金：中山大学"211工程"资助(基金号98151)

摘　　要：目的 探讨结缔组织生长因子(CTGF)在体外能否够促进肾小管上皮细胞的表型转化,以及这一作用与转化生长因子β(TGF-β)的关系。方法 将NRK52E肾小管上皮细胞株细胞分组处理,采用RT-PCR和Western印迹的方法检测CTGF mRNA水平和蛋白水平的表达;Western印迹和流式细胞仪观察α-SMA的表达。结果 正常体外培养的NRK52E肾小管上皮细胞表达少量的CTGF,加入TGF-β1 10 ng/ml刺激48h后,CTGF mRNA和蛋白水平的表达都显著升高,α-SMA蛋白表达和荧光强度也明显增加;同时加入TGF-β1中和抗体后,CTGF和α-SMA的表达都显著降低。经过反义寡核苷酸处理48h,几乎检测不到CTGF mRNA和蛋白的表达,并且CTGF反义寡核苷酸可以基本消除TGF-β1引起的细胞α-SMA表达增强,而相同时间和剂量的CTGF正义寡核苷酸不能引起相应的改变。结论 CTGF和TGF-β1都可以促使体外培养的肾小管上皮细胞α-SMA表达增强,并且CTGF作为TGF-β1的下游效应介质而起作用,提示CTGF参与了肾小管上皮细胞的转分化。Objective To investigate the significance of connective tissue growth factor (CTGF) in tubular epithelial-myofibroblast transdifferention,and the relationship between CTGF and transforming growth factor-β(TGF-β) .Methods The normal rat kidney tubular epithelial cell line(NRK52E) was cultured for 48 hours on plastic plates in the presence or absence of recombinant TGF-β1 and CTGF antisense oligonucleotide. The expression of CTGF was detected by RT-PCR and Western blot,and α-smooth muscle actin (α-SMA) was assessed by Western blot and flow eytometry. Results In NRK52E cells,basal level of CTGF expression was seen in the normal condition,Culturing in TGF-β1 10 ng/ml caused profound increase of CTGF and a-SMA level. Adding TGF-β1 neutralizing antibody largely abrogated the effect of TGF-β1. CTGF antisense oligonucleotide was showed to inhibit the expression of CTGF gene and protein,and down-regulate α-SMA expression in response to TGF-β1,but sense oligonucleotide do not. Conclusions Both CTGF and TGF-β1 are key cytokines that regulate transdifferention of tubular epithelial cells into a-SMA(+) myofiroblasts. CTGF is α TGF-β downstream mediator in this pathway.

关 键 词：结缔组织生长因子 介导转化生长因子 肾小管上皮细胞 转分化 

分 类 号：R692[医药卫生—泌尿科学]