活血解毒中药有效部位对ApoE基因敲除小鼠动脉粥样硬化斑块炎症反应的影响

Effects of Herbal Extractives with Activating Blood Circulation and Detoxication on Atherosclerotic Inflammatory Reaction in Atherosclerotic Plaque of 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基因敲除小几动脉粥样硬化（AS）斑块炎症反应的影响。方法70只（6～8）周龄小鼠ApoE基因敲除小鼠予高脂饮食喂养13周后，待其形成成熟的AS斑块后，随机分为活血解毒组（虎杖提取物组、大黄醇提物组）、活血对照组（三七总皂苷组）、解毒对照组（黄连提取物组）、模型组、阳性对照组（辛伐他汀组）。继续高脂喂养并按体重比折算的临床推荐剂量给予相应药物治疗13周后，处死动物，检测血清超敏C反应蛋白（hs—CRP）及可溶性白细胞分化抗原40配体（sCD40L）水平，并取主动脉根部4个切面，分别行HE染色和Movat染色，用免疫组化染色法观察小鼠主动脉根部斑块内过氧化物酶体增殖物激活受体-γ（PPAR-γ）的蛋白表达，实时荧光定量PCR法检测主动脉内PPAR-γ及核转录因子出（NF-κB）mRNA的表达。结果与模型组比较，大黄醇提物组及虎杖提取物组血清hs—CRP和sCD40L水平均明显降低（P〈0．01或P〈0．05）。解毒对照组黄连提取物主动脉斑块内PPAR-γ蛋白及基因表达均明显增多（P〈0．01），而中药各组小鼠主动脉斑块内NF-κB mRNA的表达与模型组比较均无统计学意义（P〈0.05）。结论采用临床推荐剂量治疗的活血解毒中药有效部位中，虎杖提取物和大黄醇提物可明显降低ApoE基因敲除小鼠血清炎症标志物hs—CRP及sCD40L水平，其效果优于单纯活血或解毒组，但对PPAR-γ和NF-κB的作用并不明显。而黄连提取物却可明显增加具有抗AS炎症作用的保护性受体PPAR-γ的蛋白和基因表达，提示其作用环节有所不同，值得深入研究。

Objective To observe the effects of several herbal extracts with activating blood circulation （saponins of Panax pseudo- ginseng）, detoxication （Coptis chinensis extractive）, activating blood circulation and detoxication （giant knotweed rhizome and rhubarb extractives） on inflammatory reaction of atherosclerotic plaques in the aortic roots of fat - fed ApoE knockout mice, Methods Seventy ApoE knockout mice were fed a high - fat diet for 13 weeks until the mature atherosclerotic plaques formed, Thereafter, they were randomized to six groups： Activating blood circulation and detoxication groups （treated with resveratrol and rhubarb extract）, activating blood circulation group （treated with notoginseng saponin）, detoxication groups （treated with coptidis rhizome extract）, control group, and positive control group （treated with simvastatin）, Thereafter, rats insix drug - treated groups were treated with respective drug and fed high - fat food for another 13 weeks, At the end of experiment, all mice were sacrificed and blood was collected for deterrninating the concentration of sCD40L and hs- CRP. The morphology and composition of atherosclerotic plaques in aortic roots were examined. The protein expression of PPAR- γ was determined by immunohistochemistry staining and the gene expression of PPAR - γ and NF - κB determined by real - time fluorescent quantitative PIER technology, Results Compared with control group, the serum concentrations of hs - CRP and sCD40L in resveratrol and rhubarb extract groups decreased significantly（P〈0. 01 or P 〈 0. 05 ）, Both the protein and gene expression of PPAR - γ in the plaque were increased significantly in coptidis rhizome extract group （P〈0. 01）, But the gene expression of NF- κB in all the medicated herbtreated groups did not change compared with controls （P 〉0. 05）, Conclusion Resveratrol and rhubarb extract could significantly decrease the concentrations of Ks- CRP and sCD40L in serum, which was superior to groups of both activating blood circulation and detoxication, but had no significant effect on the upstream inflammatory factor （NF- κB）. Coptidis rhizome extract could significantly increase the expression of PPAR - γ and suggested that there was different mechanism of action.