CTGF反义寡核苷酸对AngⅡ诱导的人近曲肾小管上皮细胞转分化的影响
人近曲肾小管上皮细胞,,]人近曲肾小管上皮细胞；血管紧张素Ⅱ；结缔组织生长因子；细胞转分化,[摘要],[关键词],1材料和方
[摘要]目的：探讨结缔组织生长因子(CTGF)反义寡核苷酸对血管紧张素Ⅱ(AngⅡ)诱导的肾小管细胞转分化的可能影响。方法：采用体外培养的人近曲肾小管上皮细胞株(HK2)，分别观察CTGF反义寡核苷酸和AngⅡ干预细胞对细胞CTGF mRNA和蛋白质的表达、细胞内超微结构及细胞α平滑肌肌动蛋白(αSMA)表达的影响。结果:AngⅡ干预HK2细胞48h后，细胞CTGF mRNA和蛋白的表达显著增高(P<0.01)；AngⅡ干预96h后，细胞超微结构显示出间质细胞样结构；这些作用都可被CTGF反义寡核苷酸显著抑制。AngⅡ可以呈时间依赖性地刺激HK2细胞表达αSMA，这些作用可以被CTGF反义寡核苷酸显著抑制(P<0.01)。对照组错义寡核苷酸无上述作用。结论：CTGF反义寡核苷酸对AngⅡ诱导HK2细胞转分化具有显著的抑制作用，提示CTGF可能介导了AngⅡ诱导的肾小管上皮细胞转分化。
[关键词]人近曲肾小管上皮细胞；血管紧张素Ⅱ；结缔组织生长因子；细胞转分化
[中图分类号]R329.28
Effect of connective tissue growth factor antisense oligonucleotide on human proximal tubular cell transdifferentiation induced by angiotensin Ⅱ
CHEN Long, LIU Bicheng, MA Kunling, LIU Hong, LUO Dongdong
(Institute of Nephrology, Zhongda Hospital, Southeast University, Nanjing 210009, China)
Abstract：Objective To investigate the effect of connective tissue growth factor antisense oligonucleotide (CTGF AS) on AngⅡ induced tubular cell transdifferentiation . Methods The influence of CTGF AS on AngⅡinduced HK2 cells CTGF mRNA and protein expression were observed by RTPCR and confocal microscopy respectively. The effect of CTGF AS on AngⅡinduced cellular ultrastructure were observed by transmissive electronic microscope (TEM). The expression of αsmooth action (αSMA) were observed by immunocytochemistry. The control group was treated with scrambled oligonucleotide (SC).Results It was shown that AngⅡsignficantly induced the increasing expression of CTGF mRNA and protein (P<0.01, respectively). If the cells were stimulated with AngⅡ for 96h, the cellular ultrastructure showed mesenchymal features. And these effects could be partially attenuated by CTGF AS. AngⅡ significantly stimulated the expression of αSMA in a timedependent manner, which could be markedly attenuated by the treatment with CTGF AS (P<0.01, respectively). The control group treated with SC had no such effects.Conclusion AngⅡ induces the EMT of tubular epithelial cells, which can be significantly attenuated by treatment with CTGF AS. Our data have provided evidence that CTGF might be involved in the AngⅡ induced tubular cell EMT. ......
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