Abstract
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 indirect-immunofluorescence. The effect of CTGF-AS on Ang II-induced cellular ultra-structure was observed using a transmissive electronic microscope. The expression of α-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 CTGF-AS (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.
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Project supported by National Natural Science Foundation of China (No 30471732), Jiangsu Provincial Natural Science Fund (BK2002052) and “135” project for Jiangsu Provincial Outstanding Medical Talent (RC2002072).
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Chen, L., Liu, Bc., Zhang, Xl. et al. Influence of connective tissue growth factor antisense oligonucleotide on angiotensin II-induced epithelial mesenchymal transition in HK2 cells . Acta Pharmacol Sin 27, 1029–1036 (2006). https://doi.org/10.1111/j.1745-7254.2006.00344.x
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DOI: https://doi.org/10.1111/j.1745-7254.2006.00344.x
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