CTGF反义寡核苷酸对人晶状体上皮细胞转分化的探讨

Influence of CTGF antisense oligonucleotide on human lens epithelial cells' trans-differentiation

目的:探讨结缔组织生长因子反义寡核苷酸对体外培养人晶状体上皮细胞(HLECs)合成CTGF与α-SMA表达的影响。方法:测定CTGF反义寡核苷酸对体外培养HLECs的转染率;应用CTGF反义寡核苷酸直接转染第3代HLECs进行细胞生长曲线的测定;采用逆转录多聚酶链式反应(RT-PCR)检测细胞CTGF和α-SMAmRNA水平。结果:直接导入法反义寡核苷酸进入较慢,但72h未出现细胞毒性。CTGF-ASON直接导入法培养细胞72h后可见对细胞增殖抑制作用。直接导入法处理48h后,TGF-β1刺激能显著增加CTGF以及α-SMAmRNA的表达,但却可被CTGF-ASON直接导入法所抑制,而错义寡核苷酸却没有这种作用。结论:CTGF反义寡核苷酸直接转染HLECs能抑制TGF-β1诱导的CTGF与α-SMA表达上调,提示阻断CTGF可能是防治后发性白内障的有效手段。

AIM：To investigate the influence of CTGF-ASON on expression of CTGF and α-SMA induced by transforming growth factor-β1（TGF-β1） in normal human lens epithelial cells（HLECs）.METHODS：CTGF-ASON was transduced into normal HLECs by cotransfection with lipofectamine 2000 or direct ways.The transfection ratio of FITC-labeled ASON was measured by fluoroscope and the proliferation of HLECs was measured by CCK-8 assay.The expression of CTGF and α-SMA was detected by RT-PCR.RESULTS：Direct ASON transfection was slow,and no obvious cytotoxicity was observed within 72 hours.The proliferation of HLECs was not inhibited by direct ASON transfection until 72 hours.ASON treated for 48 hours,Culture in TGF-β1 caused profound increase of CTGF and α-SMA that can be inhibited by direct ASON transfection,but not missense oligonucleotides transfection.CONCLUSION：CTGF-ASON can abrogate the influence of TGF-β1 on increase of CTGF and α-SMA mRNA in HLECs.CTGF may be a potential therapeutic target to posterior capsular opacification.