【作者】 李继庭；

【导师】 左俊岭；

【作者基本信息】 广州中医药大学 ， 中医内科学， 2011， 博士

【摘要】 [背景]流行性感冒(简称流感)是由流感病毒引起的急性呼吸道传染病,是一种常见病和多发病,经常引起地方性流行,有时引起世界性大流行。二十世纪曾发生过五次世界性流感大流行,给人类带来了深重的灾难。流感带来的损失不容忽视,根据美国科学技术委员会的报告,近年来科技发达的美国每年死于流感的人数为2万-4万,流感成为致死率较高的病毒性传染病；每年用于预防流感的费用和流感引起的劳动力损失超过1.2亿美元。可见流感至今仍是严重危害人类生命和健康的一种传染病。现代医学在研制抗流感病毒的药物和疫苗方面取得了长足的进步。但流感病毒具有不断发生抗原性变异的特点,抗流感病毒的药物和疫苗又具有一定的局限性,并具有不同程度的副作用。因此,目前尚无理想的防治流感的药物,研究并控制流感的工作任重而道远。流感属于中医的时行感冒范畴,为外感热病的一种。中医在流感的防治方面积累了丰富的经验,具有独特的优势,成为目前防治流感的重要研究方向。在此背景下,本研究综合伤寒论六经辨证理论及温病学卫气营血、三焦辨证理论,进行了寒温并用中药复方银翘柴桂颗粒治疗流感的理论、临床及实验研究。[目的]通过动物实验探讨银翘柴桂颗粒在调节免疫、保护脏器、抑制病毒等三方面的作用机制,评价其安全性、有效性,并通过临床实验观察该药物的疗效、症状积分改善、体温下降曲线、外周血细胞等相关指标以进一步了解其临床实用性、安全性及有效性,为银翘柴桂颗粒的临床应用提供实验依据。动物实验研究[方法]将NIH小鼠随机分为正常对照组、病毒模型组、金刚烷胺组、中药高、中、低剂量组。各组小鼠在乙醚轻度麻醉下,正常对照组用0.9%生理盐水,其余各组用甲型流感病毒FM1稀释液滴鼻以建立流感病毒感染模型。用药组分别以中药高、中、低剂量及金刚烷胺进行干预,然后根据各部分的具体实验安排进行相关指标检测。主要包括实验：1.急性毒性实验；2.银翘柴桂颗粒相关药效学研究(包括肺指数及死亡保护作用研究)；3.银翘柴桂颗粒对流感病毒FM1感染小鼠巨噬细胞吞噬功能的影响。[结果](1)急性毒性实验：各用药组小鼠无死亡,实验所取各剂量的银翘柴桂颗粒,无明显致毒性,药物安全。(2)对流感病毒FM1感染小鼠肺炎抑制作用实验：病毒感染组的肺指数较正常对照组明显增高,有显著性差异(P<0.05)。与病毒模型组相比,金刚烷胺组、中药高中低各剂量组的肺指数降低,均有显著性差异(P<0.05),中药各组的剂量与肺指数呈负相关、与肺抑制率呈正相关。(3)对流感病毒FM1感染小鼠的死亡保护作用：与病毒模型组相比,中药高、中剂量组生存时间延长,具有显著性差异(P<0.05)；中药高、中、低剂量组对流感病毒感染小鼠的死亡保护率分别是35%、25%、15%；延长生命率是38.06%、32.26%、25.81%；金刚烷胺组对流感病毒感染小鼠的死亡保护率是60%；延长生命率是43.31%；(4)对流感病毒FM1感染小鼠巨噬细胞吞噬功能的影响：病毒模型组OD值最低,与正常对照组比较,P<0.01,差异显著,说明病毒感染小鼠后其巨噬细胞吞噬功能明显降低；各药物治疗组OD值均较病毒模型组为高,P值均<0.01,差异极显著。[结论]以上结果说明银翘柴桂颗粒能有效抑制流感病毒感染小鼠的肺部炎症改变,并能延长其生存时间,具有明显的死亡保护作用,同时可以提高病毒感染后小鼠巨噬细胞的吞噬功能,起到调节免疫功能的作用。临床研究[方法]以西药抗病毒药奥司他韦及金刚烷胺为对照组药物,对银翘柴桂颗粒进行前瞻性、随机、对照研究。对142例流感患者的症状、体征、不同时间点的体温变化、疾病疗效、中医证候疗效、症状积分改善等指标进行临床观察。[结果](1)银翘柴桂颗粒在不同时间体温变化率均大于西药组,差异有显著统计学意义(P<0.01)；而且中药大剂量组与中药常规剂量组体温变化率比较有显著统计学差异(P<0.01),对三组体温变化曲线进行比较,经球形检验,差异均有统计学意义(P<0.05)。说明银翘柴桂颗粒的退热效果优于西药组,且大剂量银翘柴桂颗粒退热效果更佳；(2)银翘柴桂颗粒的症状积分改善率大于西药组,差异有显著统计学意义(P<0.01),而中药大剂量组与常规剂量组的症状积分改善率无统计学意义(P>0.05)。银翘柴桂颗粒对流感整体症状的改善明显优于西药组,而银翘柴桂颗粒在改善流感整体症状方面的量效关系未显示出统计学意义；(3)银翘柴桂颗粒两组治疗后外周血白细胞总数、中性粒细胞数较疗前降低,差异有统计学意义(P<0.05),但白细胞总数及中性粒细胞数仍在正常参考值范围内变化。银翘柴桂颗粒两组疗后与疗前相比,淋巴细胞绝对值有升高趋势,差异均有显著统计学意义(P<0.01)。(4)银翘柴桂颗粒在疾病疗效、证候疗效、主症疗效均显著优于西药组,差异有统计学意义(P<0.05),银翘柴桂颗粒的整体疗效明显优于西药组。[结论]银翘柴桂颗粒具有显著的临床疗效,尤其在退热、改善流感症状方面作用显著,并具有调节免疫的功能。更多还原

【Abstract】 ObjectiveInfluenza is a common frequent acute respiratory tract infectious disease caused by Influenzal Virus (Ⅳ) which rages in some places even around the world. There were five big rages worldwide in the 20th century that destroyed seriously. Losses caused by flu deserve anything but ignoring. According to the report of American National Science and Technology Council, recently about 20,000 to 40,000 people die of flu annually in U. S. Influenza has been a viral infectious disease with high mortality rate. Expenses for preventing flu and labor loss due to flu amount to over 120 million dollars each year. Flu is still an infectious disease, formidably threatening to human health and life.There is great advance in the anti-virus study in medicine and vaccine in the field of western medicine. However, for the continuous variation of antigen of bird-flu, the limitations and the side effect of the anti-virus medicine and vaccine in some degree, the perfect method to prevent and cure flu has not come out yet. The research for controlling flu has a long way to go.Flu belongs to "epidemic cold" in traditional Chinese medicine (TCM) theory, one of exogenous febrile diseases. So far the doctors of TCM have accumulated rich experiences in prevention and curing for eternal febrile diseases by different ways with advantages. Then the paper studied the way of treating influenza in basis of unification of "Differentiation of Six Channels" and "Differentiation of Four Systems(wei, qi, ying, and xue)" and "Differentiation of Tri-energizer".Then in order to study the mechanism and pharmacology of cold-warm combinational compound recipe Yinqiaochaigui granules treating influenza, some experiments are did.Experimental research were done for investigating the mechanism and evaluating the security and effectiveness of Yinqiaochaigui granules on regulation of the immune, protection of the organs and inhibition of the virus. Clinical research were done by observing the effect, the improvement of symptom scores, the curve of temperature drops, blood cells to clear about Yinqiaochaigui granules’ practical, security and effectiveness. So it will provide the experimental evidence for the clinical application of Yinqiaochaigui granules.Experimental Research MethodsTo study the mechanism of Yinqiaochaigui granules to cure influenza, we did some experiments. This experiments takes mouse infected by influenza virus dropping nose as the model and uses Yinqiaochaigui granules to carry out experiments. The NIH into the mice is randomly divided into nine groups:normal control group,model control group, Amantadine group, the group on high dose of traditional Chinese medicine, the group on medium dose of traditional Chinese medicine, the group on low dose of traditional Chinese medicine.There are 10 mice in each group. Under slight anesthesia caused by ether, the mice are infected by dropping influenza virus into the nose. After the day the virus attacks, medicine is given. Then, according to the objective of every expeiment, relevant indexes are measured. The experiments included:1. Acute toxicity of Yinqiaochaigui granules;2.The pharmacodynamics study including the lung exponent and life protection;3. Study of Yinqiaochaigui granules’ influence on the phagocytizing function of IV FMi infected mice’s macrophages.Result(1) Acute toxicity of Yinqiaochaigui granules:There is no mice died. The traditional Chinese medicine low/medium/high dose have no toxicity.(2)Study of Yinqiaochaigui granules’influence on the inhibition of the IV FM1 infected mice’s pneumonia:compared with normal control group,the lung exponent of the model group increase obviously which have significant difference(P<0.05). Compared with the model group,the lung exponent of amantadine group and traditional Chinese medicine low/medium/high dose group all reduced which have significant difference(P<0.05). Besides, the correlation of lung exponent and the medicine dose are negative, inhibition rate of lung and the medicine are positive.(3) Study of Yinqiaochaigui granules, influence on the protection of theⅣFM1 infected mice, s death:Compared with model group, the average life time of the FM1 infected mice in traditional Chinese medicine of high/medium/low dose groups all improved obviously;The life protection rate of the FM1 infected mice in traditional Chinese medicine of high/medium/low dose groups were 35%,25%,15%. The health extension rate of the FM1 infected mice in traditional Chinese medicine of high/medium/low dose groups were 38.06%, 32.26%,25.18%. The life protection rate and the health extension rate of the FM1 infected mice in amantadine group were 60%,43.31%.(4) Study of Yinqiaochaigui granules’influence on the phagocytizing function ofⅣFM1 infected mice’s macrophages:Compared with normal control group, the OD(optical delnsity) of model group was lower (P<0.05) which indicate the phagocytizing function ofⅣFM1 infected mice’s macrophages reduced obviously;Compared with model group, the OD of medication groups were higher(P<0.01).ConclusionSummary, Yinqiaochaigui granules can alleviate the pathological changes in the lung of FM1 infected mice and have a protective and life extension effect on FM1 infected mice. Meanwhile, Yinqiaochaigui granules can improve the phagocytizing function ofⅣinfected mice’s macrophages so that have the regulative funcation of the immune.Clinical Research MethodsThis research uses the forward-looking clinical stochastic comparison plan. The selected cases collected from the Guangzhou University of Traditional Chinese Medicine First Affiliated Hospital of outpatient and emergency patients, were randomly divided into the traditional Chinese medicine high dose group, the traditional Chinese medicine conventional dose group and the pure western medicine group three groups, integrates the case number is 142 examples. The symptoms, signs, the improvement of symptom scores, the curve of temperature drops of patients were observed in the research. Result(1)Compared with the western medicine group,the changing rate of temperature in different time of Yinqiaochaigui granules’ groups were higher(P<0.01). And The traditional Chinese medicine conventional dose group and traditional Chinese medicine high dose group were more significant statistic difference (P<0.01). Comparing the curve of temperature drops of the three groups with test of sphericity, the statistic difference were all significant(P<0.05).The result indicate that the effect of defervescence of Yinqiaochaigui granules was superior to western group, high dose Yinqiaoachaigui granules was more effective.(2) The improvement rate of symptom scores of Yinqiaochaigui granules groups were higher than western group(P<0.01). But the difference between the improvement rate of symptom scores of Yinqiaochaigui granules high dose group and conventional dose group was not significant(P>0.05). Compare with the reduction of integration of flu syndromes, the Yinqiaochaigui groups were superior to the western one. There was not dose-effect relationship between the two groups of Yinqiaochaigui.(3)The WBC and Neutrophils of Yinqiaochaigui granules goups were reduced after treatment. But the change of WBC and neutrophils was in the normal range. The lymphocytes have increasing tendency after the treatment of Yinqiaochaigui granules groups, the difference was significant (P<0.01).(4)The therapeutic effect, TCM syndromes effect of Yinqiaochaigui granules groups were superior to the western group, the difference was significant (P<0.05). The integrated effect of Yinqiaochaigui granules was obviously superior to the western medicine.ConclusionYinqiaochaigui granules have the significant clinical effect, especially in the fact of defervescence and alleviating flu symptoms. And the immune regulative funcation of Yinqiaochaigui granules was confirmed.更多还原