厄贝沙坦对AngⅡ介导的肾小球系膜细胞核因子-κB活化及MCP-1表达的影响

Irbesartan regulates activation of nuclear factor-κB and expression of monocyte chemoattractant protein-1 induced by angiotensin II in mesangial cells of rats

目的观察不同剂量厄贝沙坦(Irb)对血管紧张素Ⅱ(AngⅡ)介导的肾小球系膜细胞(GMC)核因子-κB(NF-κB)活化及单核细胞趋化因子-1(MCP-1)表达的影响,探讨Irb抑制GMC炎症因子表达的可能机制。方法分离培养大鼠GMC,用10-6mol/LAngⅡ刺激后将其随机分为5组(n=4):对照组、AngⅡ刺激组、AngⅡ和3种不同浓度(10-5mol/L、10-6mol/L、10-7mol/L)的Irb共孵育组。分别采用ELISA法及半定量RT-PCR法检测细胞培养上清液中MCP-1、IκB、NF-κB的表达水平和GMC内MCP-1mRNA的表达水平,并在酶联免疫激光扫描共聚焦显微镜下观察GMC内NF-κBp65核易位情况,计算核易位阳性率。结果与AngⅡ刺激组比较,其他各组MCP-1、IκB、NF-κB水平及MCP-1mRNA表达均明显较低(P<0.05或0.01);与低浓度Irb组比较,高浓度Irb组MCP-1、IκB、NF-κB、MCP-1mRNA表达及中浓度Irb组MCP-1水平均明显较低(均P<0.05);与中浓度Irb组比较,高浓度Irb组MCP-1、IκB、NF-κB水平及MCP-1mRNA表达均明显较低(均P<0.05)。结论Irb可能是通过抑制NF-κB的活化,降低GMCMCP-1的表达以减少肾小球系膜区单核细胞的浸润,从而减轻肾小球炎症反应。

Objective To investigate the effect of Irbesartan on activation of nuclear factor - kB （NF -k B） and expressions of monocyte chemoatractant protein-1 （MCP-1） induced by angiotensin Ⅱ（AngⅡ）in mesangial cells（GMC） of rats. Methods GMCs were extracted from healthy SD rat and cultured, the GMCs were divided into five groups： control group, Ang Ⅱ group, 3 Irbesartan （Irb） groups with different Irb concentration. The protein levels of MCP-1, I K B,NF- kB in the supernatant of GMC were measured by ELISA assay. The expression of MCP-1 mRNA was measured by reverse transcription polymerase chain reaction （RT-PCR）. Inflammation related factor NF-k B p65 was observed using laser scanning confocal microscopy. Results Comparing with Ang If group, the MCP-1, I k B,NF- k B levels and MCP-1mRNA expression in other groups were significantly decreased （P〈0.05 or 〈0.01）.The inhibition effect of Irbesartan increased with the increasing concentration of Irb. Conclusion Irbesartan can inhibit the activation of NF- k B in GMC of rats induced by the Ang Ⅱ ; it also down-regulate MCP-1 expressions, which indicates that Irbesartan might be able to reduce the monocyte infiltration in the GMC and down-regulate the inflammatory reaction level in nephritis.