【作者】 刘煜德；

【导师】 黄衍寿；

【作者基本信息】 广州中医药大学 ， 中医内科学， 2005， 博士

【摘要】 动脉粥样硬化(AS)是严重危害人类健康的疾病,近几十年来其发病率呈逐渐上升的趋势,由于病因未明,中西医对该病的防治效果都不能令人满意。近年来感染与AS的相关性引起了人们的关注。 本文从文献研究和动物实验研究两方面对AS与感染的相关性进行探讨。 第一部分 文献研究 冠心病(CAD)是动脉粥样硬化导致器官病变最常见的类型,属于中医“胸痹心痛”的范畴,自《内经》始不少医家认识到其病因与“外邪”有关,但由于受汉·张仲景《金匮要略》的“阳微阴盛”病机说的影响,治疗上侧重于祛寒温通活血,没有使用清热解毒法。 近年来现代医学发现感染与AS密切相关。根据已完成的关于感染与AS相关性的动物实验研究、以及抗生素防治冠心病的临床试验,目前还不能确定感染是否能触发或加重AS。 第二部分 实验研究 1.实验目的 探讨邪毒内侵与AS的相关性及清热解毒法的干预作用。 2.实验方法 8周龄SPF级雄性C57BL/6J小鼠84只,随机分为黄芩苷高剂量组、黄芩苷低剂量组、阿奇霉素组、高胆固醇+CPn感染组、高胆固醇组、CPn感染组、正常对照组,其中CPn感染组和正常对照组喂普通饲料和水;黄芩苷高剂量组、黄芩苷低剂量组、阿奇霉素组、高胆固醇+CPn感染组、高胆固醇组喂高胆固醇饲料,正常饮水。 喂饲高胆固醇饲料1周后,黄芩苷高剂量组、黄芩苷低剂量组、阿奇霉素组、高胆固醇+CPn感染组、CPn感染组接种CPn,方法如下:以乙醚麻醉小鼠后,加样枪吸取10ul含0.5-1×10~7IFU CPn的培养液,经一侧鼻腔吸入。每周接种1次,共3次。正常对照组以同样方法经一侧鼻腔吸入10ul培养液。 最后一次接种5天后开始给药至实验结束。黄芩苷高剂量组灌服50mg/kg/d黄芩苷,黄芩苷低剂量组灌服25mg/kg/d黄芩苷,阿齐霉素组灌服5mg/kg/d阿齐霉素,高胆固醇+CPn感染组灌服等量生理盐水,CPn感染组不治疗。更多还原

【Abstract】 Atherosclerosis(AS) is a disease which do harm to people badly. And its morbidity has been rising during recent decades. Owing to its unclear pathogen, there is no satisfying therapy to prevent and cure it in both modern medicine and traditional Chinese medicine(TCM). However, the correlation between infection and AS has aroused people’s attention recently.This thesis researches into the correlation between infection and AS.Part One: literature researchCoronary artery disease(CAD) is the most common organ pathological changes which is due to AS. CAD is belonged to the range of "chest pain" in TCM. Many Chinese ancient medical scientist realized that the pathogen of "chest pain" is related to exopathogenic factors. But owing to being influenced by (Oingguiyaolue)) , ancient medical scientists mostly used the remedies for chest pain of dispelling cold, warming and promoting blood circulation. While heat-clearing and detoxicating remedy was not used.In the recent years, infection was discovered to be involved in the genesis of CAD. However, according to the result of the finished animal experiments and clinical trials, it is still uncertain whether infection initiates or deteriorates CAD.Part Two: Animal experiment research. 1. ObjectTo study the correlation between exogenous evil and AS, and the therapeutic action of heat-clearing and detoxicating remedy.2. Methods84 eight-week-old SPF male C57BL/6J mice were divided randomly into 7 groups: baicalin high dosage (50mg/kg/d) group (group I ), baicalin low dosage (25mg/kg/d) group (group II), azithromycin(5mg/kg/d) group(group III), cholesterol+CPn group(group IV), cholesterol group(group V ), CPn group(group VI), and normal group(group VII).Group VI and group VD were fed with a regular chow diet and water throughout the study. While group I , group II, group III, group IV and group V were fed a 2% cholesterol-supplemented diet and regular water.A week after being fed a 2% cholesterol-supplemented diet, group I , group II, group III, group IV and group V were inoculated with CPn at 0.5-lx10~7 IFU per inoculation. The inoculation was carried out by dropping a total volume of 10 ul inocula into one side of the mouse nose under ether. Each mouse of these groups was inoculated once weekly for 3 consecutive weeks, group VII were sham inoculated with 10 culture medium.Administration started 5 days after the last inoculation. Group I were drenched baicalin 50mg/kg/d, group II baicalin 25mg/kg/d, group III azithromycin 5mg/kg/d. Group IV, group V, group VI, group andVII were drenched aequales volume of normal saline.14 weeks after the final inoculation, 2 mice were selected randomly from each group and sacrificed for aorta isolation to observe the injured endothelial cells.18 weeks after the final inoculation, all mice were sacrificed for serum collection and the isolation of aorta, heart, lung, liver and kidney. The serum was used to test CPn antibodies, TNF-a IL-6 EL-10 sE-selectin sVCAM-land sICAM-1. Aorta was used to assess plaque area index(PAI), CD40-CD40L expression intensity and apoptosiss. The heart, lung, liver and kidney were pathologically studied.3. Rusult3.1 The effect of CPn on PAI and the intervention of medicine.Compared with group VII, PAI in group IV and group V differed significantly (p<0.01). Although PAI in group IV is a little greater compared to group V, there is no significant difference (p>0.05).PAI in treatment groups differ significantly compared with group IV and with group VII. Compared with group III, PAI in group II were smaller (p<0.05), whereas group I had no significant difference (p>0.05).3.2 The effect of CPn on inflammatory factors and the intervention of medicine.3.2.1 The effect of CPn on TNF— a and the intervention of medicine.Compared with normal group, the level of serum TNF—a in all model groups increased (p<0.01). Thereinto, TNF—a in group IV was higher than that in group V and group VI (p<0.01). Compared with group IV, TNF—a in all treatment groups decreased remarkably (p<0.01). However, only TNF—a in group I was close to that in normal group (p>0.05). There is no significant difference among treatment groups (p>0.05).3.2.2 The effect of CPn on IL—6 and the intervention of medicine.Compared with normal group, the level of serum IL—6 in group VI and group IV decreased (p<0.01), while that in group V was no significant difference (p>0.05). Compared with group IV, IL—6 in all treatment groups decreased remarkably (p<0.01), which was close to the level of normal group (p>0.05).3.2.3 The effect of CPn on IL—10 and the intervention of medicine.Compared with normal group, the level of EL—10 in group V and group VI had no significant difference(p>0.05), whereas group IV increased significantly (p<0.01). Compared with group IV, IL—10 in group I and group II decreased remarkably (p<0.01), which was close to the level of normal group (p>0.05). IL—10 in group III versus group IV did not differ (p>0.05), which versus normal group was higher remarkably (p<0.01).3.3 The effect of CPn on ICAM—1, VCAM—Is E—slectin and the intervention of medicine.Compared with normal group, the level of soluble ICAM—1 in group IV and group VI invreased significantly (p<0.01). ICAM—1 in group IV versus group VI did not differ (p>0.05). Compared with group IV, ICAM—1 in all the three treatment groups decreased some extent. But only group II and group III had significant differenceCompared with normal group, the level of soluble VCAM—1 in group IV increased remarkably (p<0.01), while that in group V and group VI had no significant changes (p>0.05). Compared with group IV, VCAM—1 in all the treatment groups decreased some extent, but they all had no significant difference (p>0.05). Compared with normal group, group II had no significant changes (p>0.05), while group I and group III increasedremarkably (p<0.01 and p<0.05 respectively).Compared with normal group, the level of soluble E—selectin in group IV and group VI increased significantly (p<0.01), while that in group V has no significant changes (p<0.01). Compared with group IV, E~selectin in all the treatment groups decreased some extent, but only group II and group III had significant difference (p<0.01), while group I had no significant changes.3.4 The effect of CPn on endothelial cell injury and the intervention of medicine.the extent of endothelial cell injury in group IVand group V increased remarkably (p<0.01), while group VI had no significant changes (p>0.05). Compared with group IV, the extent of endothelial cell injury in all the treatment groups decreased remarkably (p<0.01), but did not reach the level of normal group (p<0.01). The three treatment groups had no significant difference (p>0.05).3.5 The effect of CPn on plaque stability and the intervention of medicine.3.5.1 The effect of CPn on CD40 and CD40L expression intensity in aortic intima and the intervention of medicine.Compared with normal group, CD40 expression intensity in group IV and group V increased remarkably (p<0.01), while group VI had no significant changes (p>0.05). All the treatment groups versus group IV did not differ (p>0.05). All the treatment groups were higher than normal group (p<0.01).Compared with normal group, CD40L expression intensity in group IV and group V increased remarkably, while group VI had no significant changes (p>0.05). Compared with group IV, CD40L expression intensity in group III decreased, but did not reach the level of normal group; while group I and group II had no significant changes (p>0.05).3.5.2 The effect of CPn on apoptosis in aortic intima and the intervention of medicine.Compared with normal group, apoptosis in aortic intima in group IV and group V increased remarkably (p<0.01), while group VI had no significant changes (p>0.05). Group IV was higher than group V.All the treatment group versus group IV decreased remarkably (p<0.01), but did not reach the normal level (p<0.01). Compared to each other in the three treatment groups, they all had significant difference (p<0.01).更多还原