Influence of connective tissue growth factor antisense oligonucleotide on angiotensin II-induced epithelial mesenchymal transition in HK2 cells  

作　　者：Long CHEN Bi-cheng LIE Xiao-liang ZHANG Jian-dong ZHANG Hong LIU Min-xia LI 

机构地区：[1]Institute of Nephrology, Zhong Da Hospital, Southeast University School of Medicine, Nanjing 210009, China

出　　处：《Acta Pharmacologica Sinica》2006年第8期1029-1036,共8页中国药理学报（英文版）

基　　金：Project supported by National Natural Science Foundation of China(№ 30471732);Jiangsu Provincial Natural Science Fund(BK2002052);"135"project for Jiangsu Provincial Outstanding Medical Talent(RC2002072).

摘　　要：Aim： The present study was designed to further investigate the effect of connective tissue growth factor antisense oligonucleotide （CTGF-AS） on angiotensin Ⅱ （Ang Ⅱ）-induced tubular cell epithelial mesenchymal transition （EMT） in vitro. Methods： The human proximal tubular cell line （HK2） was grown in Dulbecco＇s modified Eagle＇s medium containing 10% heat inactivated fetal calf serum. After being rested in serum-free medium for 24 h, the influence of CTGF-AS （20 μg/mL） on Ang Ⅱ-induced （1 × 10^-7 mol/L） CTGF mRNA and the protein expression were examined by using reverse transcription-polymerase chain reaction and indirectimmunofluorescence. The effect of CTGF-AS on Ang Ⅱ-induced cellular ultrastructure was observed using a transmissive electronic microscope. The expression of or-smooth action （α-SMA） was assayed by immunocytochemistry. In all experiments, the control group was treated with scrambled oligonucleotide. Results： It was shown that Ang Ⅱ significantly induced the increasing expression of CTGF mRNA and protein （P〈0.01, respectively）, which were significantly abolished by treatment with CTGF-AS. After stimulating cells with Ang Ⅱ, the cellular ultrastructure showed mesenchymal features. These effects were partially inhibited by CTGF-AS. Ang Ⅱ significantly resulted in the expression of α-SMA in time dependent manner, which was markedly attenuated by the treatment with CTGFAS （P〈0.01, respectively）. In contrast, no similar effects were observed in the control group treated with scrambled oligonucleotide. Conclusion： Ang Ⅱ-induced EMT in human proximal tubular epithelial cells （PTC） can be attenuated by treatment with CTGF-AS. Our data provides further evidence that CTGF might be involved in Ang Ⅱ-induced EMT in PTC.Aim： The present study was designed to further investigate the effect of connective tissue growth factor antisense oligonucleotide （CTGF-AS） on angiotensin Ⅱ （Ang Ⅱ）-induced tubular cell epithelial mesenchymal transition （EMT） in vitro. Methods： The human proximal tubular cell line （HK2） was grown in Dulbecco＇s modified Eagle＇s medium containing 10% heat inactivated fetal calf serum. After being rested in serum-free medium for 24 h, the influence of CTGF-AS （20 μg/mL） on Ang Ⅱ-induced （1 × 10^-7 mol/L） CTGF mRNA and the protein expression were examined by using reverse transcription-polymerase chain reaction and indirectimmunofluorescence. The effect of CTGF-AS on Ang Ⅱ-induced cellular ultrastructure was observed using a transmissive electronic microscope. The expression of or-smooth action （α-SMA） was assayed by immunocytochemistry. In all experiments, the control group was treated with scrambled oligonucleotide. Results： It was shown that Ang Ⅱ significantly induced the increasing expression of CTGF mRNA and protein （P〈0.01, respectively）, which were significantly abolished by treatment with CTGF-AS. After stimulating cells with Ang Ⅱ, the cellular ultrastructure showed mesenchymal features. These effects were partially inhibited by CTGF-AS. Ang Ⅱ significantly resulted in the expression of α-SMA in time dependent manner, which was markedly attenuated by the treatment with CTGFAS （P〈0.01, respectively）. In contrast, no similar effects were observed in the control group treated with scrambled oligonucleotide. Conclusion： Ang Ⅱ-induced EMT in human proximal tubular epithelial cells （PTC） can be attenuated by treatment with CTGF-AS. Our data provides further evidence that CTGF might be involved in Ang Ⅱ-induced EMT in PTC.

关 键 词：connective tissue growth factor angiotensin Ⅱ epithelial mesenchymal transition proximal kidney tubules humans 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]