原发性高血压患者中抗α1肾上腺素受体自身抗体激活大鼠主动脉平滑肌细胞内 NF-κB

摘要

目的：探讨高血压患者血清中抗α1肾上腺素受体自身抗体（autoantibodies against α1-adrenergic re-ceptor，α1-AAs）拟去甲肾上腺素的激动样效应及胞内信号分子活化机制。方法：培养大鼠主动脉平滑肌细胞。将收集的原发性高血压病人血清进行免疫亲和层析法纯化，ELISA检测其滴度后以1∶80刺激细胞，并设立不同的处理组。将α1-AAs刺激大鼠主动脉平滑肌细胞后，经Western blotting及免疫荧光方法分析胞内核因子κB （nuclear factor κB， NF-κB）信号分子激活及表达状况。结果： Western bltting 显示α1-AAs 刺激组中胞内核因子NF-κB （p50）表达较空白对照组明显增强，且可被α1受体阻滞剂哌唑嗪和NF-κB拮抗剂PDTC明显抑制（P＜0．05）。免疫荧光实验中，α1-AAs组荧光强度显著高于空白对照组和α1-AAs＋哌唑嗪组（P＜0．01）。结论：高血压患者血清中α1-AAs能够致大鼠主动脉平滑肌细胞NF-κB活化。%AIM:To study the effects of autoantibodies against α1-adrenergic receptor (α1-AAs) isolated from the hypertensive patients, which showed the agonist-like activity similar to norepinephrine , on the signal mechanism of vas-cular smooth muscle cells (VSMCs) isolated from rat thoracic aorta.METHODS:Rat VSMCs were cultured and identi-fied.The serum of hypertensive patients was purified by immunoaffinity chromatography .The autoantibodies were detected by ELISA and used to activate the cells with the titer of 1∶80.The total protein was extracted and the expression of NF-κB in different treatment groups was detected by Western blotting .Meanwhile, the activation of NF-κB in the nucleus was ana-lyzed by immunofluorescence method .RESULTS: The expression of NF-κB in VSMCs was obviously higher in α1-AAs group than that in control group .Meanwhile, the expression of NF-κB was inhibited by prasozin and PDTC.The autoanti-bodies caused a significant increase in NF-κB expression in the nucleus .The fluorescence intensity in α1-AAs group was high than that in control group and α1-AAs+prasozin group (P<0.01).CONCLUSION:Theα1-AAs from hypertensive patients increase NF-κB expression in rat VSMCs.