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目的检测血管紧张素II(Ang II)介导的大鼠血管平滑肌细胞(VSMC)增殖过程中信号转导和转录活化因子-1(STAT1)的激活与核转位。方法本文采用Western印迹、非同位素凝胶电泳(EMSA)和免疫荧光染色的方法,观察Ang II刺激大鼠主动脉VSMC前后,细胞中STAT1的活化状态与定位。结果VSMC经Ang II干预后,胞内磷酸化的STAT1(p-STAT1)蛋白表达增加(P<0.01),达峰后随时间梯度逐渐下降,Ang II干预15 min后检测到胞核内有蛋白-DNA复合物形成。这一反应可被血管紧张素II 1型受体(AT1)阻滞剂Losartan以及Jak2抑制剂AG490抑制(P<0.01)。免疫荧光染色结果也显示Ang II干预后p-STAT1主要在胞核内表达。结论Ang II可以通过和AT1受体结合,激活Jak/STAT通路,在Ang II介导的大鼠VSMC增殖过程中发挥作用。
Objective To investigate the activation and nuclear translocation of signal transducer and activator of transcription 1 (STAT1) in the proliferation of rat vascular smooth muscle cells (VSMCs) induced by angiotensin II (Ang II). Methods Western blotting, non-isotope gel electrophoresis (EMSA) and immunofluorescence staining were used to observe the activation status and localization of STAT1 in the cells before and after Ang II stimulation. Results The expression of STAT1 (p-STAT1) protein in VSMCs was increased after Ang II intervention (P <0.01), and gradually decreased with time gradient after reaching the peak. After 15 min of Ang II intervention, protein -DNA complex formation. This response was inhibited by Losartan, an angiotensin II type 1 receptor (AT1) blocker, and AG490, a Jak2 inhibitor (P <0.01). Immunofluorescence staining also showed that p-STAT1 was mainly expressed in the nucleus after Ang II intervention. Conclusion Ang II can activate Jak / STAT pathway by binding to AT1 receptor and play an important role in the proliferation of rat VSMC induced by Ang II.