摘要
目的探讨一氧化氮—环磷酸鸟苷依赖性蛋白激酶信号通路是否通过对GATA-6表达进行调节来抑制苯肾上腺素诱导的血管平滑肌细胞表型转化。方法组织贴块法原代培养Wistar大鼠血管平滑肌细胞。四甲基偶氮唑盐法测定血管平滑肌细胞的增殖,逆转录聚合酶链反应以及免疫印迹法测定GATA-6和平滑肌肌球蛋白重链的表达水平。结果一氧化氮供体S-亚硝基-N-乙酰青霉胺和环磷酸鸟苷依赖性蛋白激酶选择性环磷酸鸟苷类似物Sp-8-pCPT-cGMPs抑制苯肾上腺素诱导的血管平滑肌细胞增殖,但环磷酸鸟苷依赖性蛋白激酶拮抗剂Rp-8-pCPT-cGMPs促进血管平滑肌细胞增殖。S-亚硝基-N-乙酰青霉胺和Sp-8-pCPT-cGMPs促进GATA-6和平滑肌肌球蛋白重链mRNA和蛋白质的表达,而Rp-8-pCPT-cGMPs则降低其表达水平。结论一氧化氮—环磷酸鸟苷依赖性蛋白激酶信号通路在转录水平和翻译水平促进GATA-6和平滑肌肌球蛋白重链的表达,进而参与维持血管平滑肌细胞分化表型。
Aim To discuss the regulation of expression d GATA-6 by nitric oxide/cGMP-dependent protein kinase I in vascular smooth muscle cells (VSMC) phenotypie modulation. Methods Cultured wistar rat aortic VSMC were used as an experimental model. Cell growth was determined by MTr assay. The nRNA and protein expression of GATA-6 and smooth muscle myosin heavy chain(sm-MHC) were assayed by RT-PCR and Western blot analysis. Results The results showed that phenylephrineinduced proliferation of VSMC was inhibited by S-nitroso-N-Acetylpenicillamine (SNAP) and Sp-8-pCPY-cGMPs, but promoted by Rp-8-pCPT-cGMPs. The expression of GATA-6 and sm-MHC mRNA and protein were increased by SNAP and Sp-8-pCPT-cGMPs, and decreased by Rp-8-pCPY-cG-MPs. Conclusions NO/PKG I can up-regulate the expression of GA- TA-6 and sm-MHC at the transcriptional and translational level. These findings demonstrate that NO/PKG I pathway is involved in the maintenance of the differentiated phenotype in VSMC.
出处
《中国动脉硬化杂志》
CAS
CSCD
2007年第9期687-690,共4页
Chinese Journal of Arteriosclerosis