原发性高血压患者中抗α_1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

Activation of NF-κB in rat vascular smooth muscle cells by autoantibodies against α_1-adrenergic receptor from hypertensive patients

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 againstα1-adrenergic receptor,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 factorκB,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。

[ ABSTRACT] AIM： To study the effects of autoantibodies against ctl-adrenergic receptor （ctl-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-KB in different treatment groups was detected by Western blotting. Meanwhile, the activation of NF-KB in the nucleus was ana lyzed by immunofluorescence method. RESULTS： The expression of NF-KB in VSMCs was obviously higher in otl-AAs group than that in control group. Meanwhile, the expression of NF-KB was inhibited by prasozin and PDTC. The autoanti bodies caused a significant increase in NF-KB expression in the nucleus. The fluorescence intensity in αl-AAs group was high than that in control group and αl-AAs ＋ prasozin group （P 〈0.01 ）. CONCLUSION： The αl-AAs from hypertensive patients increase NF-KB expression in rat VSMCs.