目的观察血性脑脊液对大鼠脑动脉平滑肌细胞蛋白激酶GIamRNA及其蛋白表达水平变化与增殖的关系,探讨蛋白激酶GIa基因在蛛网膜下腔出血后继发慢性脑血管痉挛分子机制中的调控作用。方法组织块法原代培养脑动脉平滑肌细胞。采用半定量逆转录多聚酶链反应技术检测对照组、血性脑脊液处理24h组、血性脑脊液处理48h组蛋白激酶GIa基因mRNA的表达水平;采用蛋白质印迹技术检测相应的蛋白的表达水平。采用甲基噻唑基四唑法、氚标记胸腺嘧啶核苷掺入法检测脑动脉平滑肌细胞增殖改变。结果血性脑脊液诱导脑动脉平滑肌细胞不断增殖。各组均检测出蛋白激酶GIa基因的mRNA以及蛋白表达的变化,图像定量分析光密度值显示各组表达的mRNA以及蛋白表达与细胞增殖改变密切相关。结论蛋白激酶GIa基因可能在蛛网膜下腔出血后脑血管痉挛中调节脑动脉平滑肌细胞增殖。
Aim To observe the changes of PKGIa mRNA and protein expressive level in cerebral arterial smooth muscle cells(CASMC)induced by bloody cerebrospinal(BCSF)in rats,and to investigate the value of PKGIa signaling pathway in cerebral vasospasm(CVS)after subarachnoid hemorrhage(SAH).Methods The pure CASMC were cultured by tissue-sticking methods.Semi-quantitative reverse transcription and polymerase chain reaction(RT-PCR)were used to examine the PKGIa mRNA expression after CASMC were exposed to BCSF for 24 h and 48h respectively.And PKGIa protein expressions were determined by Western blotting.The changes of CASMC proliferation were determined by MTT and ^3H-TdR incorporated way.Results BCSF could stimulate CASMC proliferating.The mRNA and protein of PKGIa could be detected in all groups,and the expressive level of mRNA and protein had a possible relationship with CASMC prolferation.Conclusion The results suggested that PKGIa signaling pathway might play an important role in the CVS after SAH.