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Aim:The present study was designed to further investigate the effect of connec-tive 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’smodified Eagle’s medium containing 10% heat inactivated fetal calf serum.Afterbeing 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 wereexamined 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 expres-sion of or-smooth action(α-SMA)was assayed by immunocytochemistry.In allexperiments,the control group was treated with scrambled oligonucleotide.Results:It was shown that Ang Ⅱ significantly induced the increasing expressionof CTGF mRNA and protein(P<0.01,respectively),which were significantly abol-ished by treatment with CTGF-AS.After stimulating cells with Ang Ⅱ,the cellularultrastructure showed mesenchymal features.These effects were partially inhib-ited by CTGF-AS.Ang Ⅱ significantly resulted in the expression of α-SMA in timedependent manner,which was markedly attenuated by the treatment with CTGF-AS(P<0.01,respectively).In contrast,no similar effects were observed in thecontrol group treated with scrambled oligonucleotide.Conclusion:Ang Ⅱ-in-duced EMT in human proximal tubular epithelial cells(PTC)can be attenuated bytreatment with CTGF-AS.Our data provides further evidence that CTGF might beinvolved in Ang Ⅱ-induced EMT in PTC.
Aim: The present study was designed to further investigate the effect of connec-tive tissue growth factor antisense oligonucleotide (CTGF-AS) on angiotensin II (Ang II) -induced tubular cell epithelial mesenchymal transition (EMT) in vitro. Methods: The human The proximal tubular cell line (HK2) was grown in Dulbecco’s modified Eagle’s medium containing 10% heat inactivated fetal calf serum. After be be 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 wereexamined 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 expres-sion of or-smooth action (α-SMA) was assayed by immunocytochemistry. In allexperiments, the control group was treated with scrambled oligonucleotide. Results: It was shown that Ang Ⅱ significantly induced the increasing expressionof CTGF mRNA and protein (P <0.01, respectively), which were significantly abol-ished by treatment with CTGF-AS. Afterfter stimulating cells with Ang II, the cellularultrastructureevenous mesenchymal features.These effects were partially inhib- ited by CTGF-AS. Ang Ⅱ RTI> resulting in the expression of a-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-in-duced 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 in isolated in Ang II-induced EMT in PTC.