小檗碱对颈动脉粥样硬化家兔NF-κB、MCP-1表达的影响

The effect of berberine on the NF-κB and MCP-1 in rabbits with carotid artery atherosclerosis

目的探讨小檗碱预防颈动脉粥样硬化形成的作用机制。方法将24只雄性新西兰大白兔随机分为3组:正常组、颈动脉粥样硬化组(模型组)、小檗碱预防组(小檗碱组),每组各8只;正常组给予普通饮食5周;模型组和小檗碱组给予高脂饲料喂养1周后行右侧颈动脉内膜空气干燥术,术后继续高脂饲料喂养4周以制成颈动脉粥样硬化模型;小檗碱组在给予高脂饲料喂养的同时每日灌服小檗碱;第5周取右侧颈动脉组织行HE染色,并观察颈动脉病理改变;行核因子-κB(Nuclear factor kappa B,NF-κB)免疫组织化学染色半定量检测NF-κB的活性,用RT-PCR法检测NF-κBp65mRNA、MCP-1mRNA表达水平。结果模型组颈动脉血管内膜明显增生,内膜下和中膜可见大量泡沫细胞堆积,有明显的动脉粥样硬化斑块形成;小檗碱组的内膜轻度增厚,内膜下有少量泡沫细胞形成;小檗碱组NF-κBp65阳性细胞数密度高于正常组(P<0.01),明显低于模型组(P<0.01);小檗碱组NF-κBp65/β-actin、MCP-1/β-actin扩增带吸光度值比值高于正常组(0.42±0.05vs0.18±0.04,0.57±0.18vs0.33±0.49,P<0.01),而明显低于模型组(0.42±0.05vs0.66±0.10,0.61±0.11vs0.76±0.03,P<0.01);NF-κB活性与MCP-1呈显著正相关(r=0.958P<0.01)。结论小檗碱预防家兔颈动脉粥样硬化可能通过抑制NF-κB活性,降低MCP-1表达而起作用。

Objective To investigate the mechanism of berberine on preventing atherosclerosis （AS） in rabbits. Methods New Zealand white rabbits （n = 24） were randomly divided into three groups including the normal group,the carotid artery atheroselerosis group and berberine group （n = 8）. The rabbits of the normal group were fed with normal diet. Carotid atherosclerosis was established by an air-drying operation which was performed on the right common carotids, and in conjunction with a high cholesterol-diet in the second week for 4 weeks. Berberine was administrated intragastricly daily from the first week for 5 weeks. The right carotid arteries were assayed by Hematoxylin and Eosin （HE） staining to investigate the effect of berberine on carotid atherosclerotic lesion, and by immunohistochemistry staining to assay the effects of berberine on the activity of nuclear factor kappa B （NF-κB）,and by RT-PCR to assay the effects of berberine on the expression of mono cyte chemoattractant protein-1 and Nuclear factor kappaB p65. Results The air-operation in conjunction with a high cholesterol diet increased numberous foam cells compared with the normal group, in which were decreased in the berberine group. The relative copies of MCP-1 mRNA expression on β-actin increased from 0. 33 ±0. 05 （the normal group） to 0. 76 ±0. 03 （the carotid artery atherosclerosis group） （P〈0. 01 ）, and decreased to 0. 57 ±0. 18 （berberine group）. The relative copies of NF-κBp65mRNA on β-actin increased from 0. 18 ±0. 04 （the normal group） to 0. 66 ±0. 16 （the carotid artery atherosclerosis group） （P〈0. 01）, and decreased to 0. 42 ±0. 05 （berberine group） （P〈0. 01）. The activity of NF-κB was positive associated with the expression of MCP-1 mRNA （r = 0. 958,P〈0. 01）. Conclusions Berberine could prevent carotid artery atherosclerosis by inhibiting the activity of NF-κB and the expression of MCP1.