活血解毒中药有效部位对ApoE基因敲除小鼠血脂和动脉粥样硬化斑块炎症反应的影响

Effects of Some Active Ingredients of Chinese Drugs for Activating Blood Circulation and Detoxicating on Blood Lipids and Atherosclerotic Plaque Inflammatory Reaction in ApoE-gene Knockout Mice

目的观察活血(三七总皂苷)、解毒(黄连提取物)、活血解毒中药有效部位(虎杖提取物、大黄醇提物)对ApoE基因敲除小鼠血脂和主动脉粥样斑块炎症反应的影响。方法ApoE基因敲除小鼠予高脂饮食喂养26周,给药13周后,处死检测血脂,并取出心脏及主动脉,石蜡包埋,取主动脉根部4个切面,分别行HE染色、Movat染色,测量并计算脂质核心占斑块总面积的百分比以及斑块内脂质成分/胶原成分比值;每个样本取4个切面的平均值。免疫组织化学染色法观察小鼠主动脉根部粒细胞/巨噬细胞集落刺激因子(GM-CSF)、肿瘤坏死因子α(TNF-α)的表达。结果给药13周后,斑块内脂质核心面积、斑块内脂质成分/胶原成分比值,中药各给药组与模型组比较均有不同程度降低(P<0.01),其中活血解毒中药虎杖提取物降低最为显著,并与活血中药三七总皂苷组、解毒中药黄连提取物组比较亦具有显著差异(P<0.05)。虎杖提取物、黄连提取物、三七总皂苷均具有程度不同的调脂作用,但仍以虎杖提取物的作用最为显著;各给药组只有大黄醇提物组的TNF-α表达减少,与模型组比较差异有统计学意义(P<0.05),中药各组GM-CSF表达减少,与模型组比较,差异有统计学意义(P<0.05)。结论活血、解毒及活血解毒中药有效部位三七总皂苷、黄连提取物、虎杖提取物、大黄醇提物在临床推荐剂量上均可通过改善斑块内部成分来稳定易损斑块,其中,兼有活血和解毒作用的中药虎杖提取物、大黄醇提物效果最为显著。其机制可能与调节脂质代谢和抑制炎症反应有关。

Objective To observe the effects of active ingredients from Chinese drugs for activating blood circulation and detoxicating, including notoginseng saponins （ drug 1 ）, Coptis chinensis （ drug 2 ）, giant knotweed rhizome （ drug 3 ） and rhubarb （ drug 4 ）, on blood lipids and inflammatory reaction of aortic atherosclerotic plaques in ApoE knockout mice. Methods ApoE knockout mice were fed with high-fat diet for 26 weeks, then they were randomized into 6 groups, the untreated model group and the test groups treated with various test drugs respectively. After ending the 13 weeks of treatment, all the mice were sacrificed with their blood lipids detected, and their heart and aorta were taken out to make slices with paraffin embedding. Four sections from aortic root of each mouse were chosen to measure and calculate the percentage of lipid core （LC） in the total area of plaque （TP） and the lipid/collagen ratio （L/C） in the plaque by HE and Movat staining respectively, and the mean value of the four sections was taken for analysis. The expressions of granulocyte-macrophage colony-stimulating factor （GM-CSF） and tumor necrosis factor-α （TNF-α） in mice＇ s aorta root were determined by immunohistochemical staining as well. Results After being treated for 13 weeks, either the percentage of LC in TP and the L/C ratio was significandy lower in all the test drug treated groups than those in the model group, respectively （P 〈0. 01 ） , especially prominent in the group treated with drug 3. Although lowering of the two indexes presented in all the 3 groups treated by drug 1, 2 and 3, significant difference still presented between drug 3 treated group vs drug 1 and 2 treated group （P 〈 0. 05 ）. As for the expressions of GM-CSF and TNF-α, in comparing with the untreated model group, significant decreasing of the TNF-α showed only in the drug 4 treated group, while that of GM-CSF could be found in all the test drug treated groups （ P 〈 0. 05 ）. Conclusion All the 4 drugs tested in the recommended dosage can stabilize the vulnerable plaques in ApoE knockout mice by improving the constitution of plaque, among them, drug 3 and 4, the drugs possess both the actions of activating blood circulation and detoxicating, show more significant effect, and their mechanisms may be related to their actions in regulating lipid metabolism and inhibiting inflammatory reaction.