谢健　沈超　童晓云　李晓　石安华　陈文玲　叶勇　刘涛

摘要：目的 探讨活血通脉颗粒干预颈动脉粥样硬化（carotid atherosclerosis，CAS）斑块形成的机制及对炎症因子表达的抑制作用。方法 将84只兔随机抽取10只为空白组，其余采用高脂饲料喂养建立CAS模型。将成模48只CAS兔随机分为模型组、自然消退组、辛伐他汀组和活血通脉低、中、高剂量组，每组8只。各给药组给予相应药物灌胃，每日1次，连续5周。取颈动脉组织行常规HE染色观察病理变化；免疫组化检测兔颈动脉TLR4的活性表达；荧光定量PCR检测各组TLR4及炎症因子单核细胞趋化蛋白-1（MCP-1）、核因子（NF）-κB mRNA表达。结果 模型组颈动脉内膜明显增厚，动脉粥样硬化分布广泛，斑块内含有大量泡沫细胞；免疫组化染色各给药组TLR4阳性染色面积百分比明显低于自然消退组（P<0.01）；活血通脉高剂量组与辛伐他汀组比较无明显差异；荧光定量PCR检测TLR4、NF-κB、MCP-1 mRNA表达各给药组低于自然消退组（P<0.05）。结论 活血通脉颗粒可能通过TLR4信号通路干预CAS模型兔斑块形成及对炎症因子表达的抑制作用达到治疗目的。

关键词：活血通脉颗粒；颈动脉粥样硬化；TLR4信号通路；单核细胞趋化蛋白-1；核因子-κB；兔

DOI：10.3969/j.issn.1005-5304.2016.03.013

中图分类号：R285.5 文献标识码：A 文章编号：1005-5304（2016）03-0047-04

Abstract： Objective To discuss the mechanism of rabbit plaque formation of carotid atherosclerosis （CAS） and inhibition of the expression of inflammatory factors. Methods Ten white rabbit randomly selected from 84 white rabbits were set as blank group， and the rest rabbits were fed with high-fat diet for the establishment of CAS models. 48 CAS model rabbits were randomly divided into model group， the natural healing group， simvastatin group， and Huoxue Tongmai Granule low-， medium- and high-dose groups， 8 rabbits in each group. Each medication group was given relevant medicine for gavage， once a day for 5 weeks. Carotid artery tissue routine HE staining was used to observe the pathological changes. Immunohistochemical detection was used for the activity of TLR4 expression in rabbit carotid artery. Fluorescence quantitative PCR was used to detect the TLR4 and the inflammation factor MCP-1， NF-κB mRNA expression. Results The rabbit endometria in model group were obvious thickening； atherosclerosis was widely distributed； a large number of foam cells in plaque were observed. Immunohistochemical in treatment groups showed that TLR4 positive staining area percentage significantly smaller than the natural healing group （P<0.01）. There were relatively few differences between Huoxue Tongma Granule high-dose group and simvastatin group. The expressions of TLR4， NF-κB， MCP-1 mRNA in the treatment group were lower than the natural healing group by Fluorescence quantitative PCR detection （P<0.05）. Conclusion Huoxue Tongmai Granules achieve therapeutical effect by intervening CAS rabbit plaque formation and inhibiting the expression of inflammatory factors through TLR4 signaling pathways.

Key words： Huoxue Tongmai Granules； carotid atherosclerosis； TLR4 signaling pathways； MCP-1； NF-κB； rabbits

随着人口老龄化加快，颈动脉粥样硬化（carotid atherosclerosis，CAS）发病率不断上升，炎症反应贯穿CAS发病的始终。临床应用活血通脉颗粒治疗CAS，疗效较好[1-2]，但其具体作用机制仍未明确。本实验观察活血通脉颗粒是否通过TLR4抑制炎症因子，从而达到抑制炎症反应、延迟斑块形成、稳定斑块、治疗CAS的作用。

1 材料与方法

1.1 药物

活血通脉颗粒由红景天、虎杖、三七粉、水蛭、茯苓、隔山消等组成，上述饮片均购于云南省中医医院中药房，并由该院制剂中心配制。辛伐他汀片，默沙东制药有限公司，规格40 mg/片，批号K005133。

1.2 动物与饲料