摘要
目的探讨蛋白激酶C在1-磷酸鞘氨醇(Sphingosine 1-phosphate,S1P)激活大鼠心肌细胞核转录因子(Nuclear transcription factor,NF)-κB信号传导通路中的作用。方法心肌细胞NF-κB的核内浓度采用夹心ELISA的方法检测;免疫印记观察S1P作用前后核内NF-κB的改变;RT-PCR及免疫印记检测转化生长因子β1(transforming growth factorβ1,TGF-β1)变化。结果 S1P刺激可诱导心肌细胞下游NF-κB活化,两者之间存在浓度与时间的依赖关系;免疫印记显示S1P作用后,细胞核内NF-κB表达明显增加;而特异性PKC抑制剂(Staurospo-rine)和NF-κB阻断剂(PDTC)均能在不同程度上抑制S1P的这种作用。S1P作用心肌细胞使TGF-β1表达上调,S1P的这种作用可被Staurosporine和PDTC所抑制。结论 PKC作为NF-κB活化的上游信使,参与了S1P激活NF-κB的信号传导通路。通过以上途径使TGF-β1合成增加,从而诱导心肌细胞纤维化,导致心肌肥厚。
Objective To study the roles of protein kinase C （PKC） in the signal transduction pathway of S1 P induced nuclear transcription factor （NF） -κB activation of cardiomyocyte cells. Method NF-κB level in nuclear protein extraction of cardiomyocyte cells was detected by sandwich ELISA. Intranuclear translocation of NF-κB was observed by WB. TGF-β1protein and RNA level was detected by WB and RT-PCR. Result SIP could induce NF-κB activation of cardiomyocyte cells in time and dose dependent manners. WB revealed intranuclear translocation following S1P induction. Specific PKC （Staurosporine） and pyrrotidine dithiocarbamate （PDTC） could inhibit SIP-induced NF-κB activation in cardiomyocyte cells. The up-expresssion of TGF-β1 by SIP was also inhibited by Staurosporine and PDTC. Conclusion PKC, as an up-stream messenger, is involved in the signal transduction pathway of S1 P-induced NF-κB activation.
出处
《中南医学科学杂志》
CAS
2011年第6期629-633,共5页
Medical Science Journal of Central South China
关键词
蛋白激酶C
1-磷酸鞘氨醇
核因子ΚB
信号传导
protein knase C
sphingosine-1-phosphate
nuclear transcription factor -κB
signal transduction
