温病透泄热郁法的研究

发布时间：2018-06-13 08:18

本文选题：温病透泄热郁法 + 方证理论　；参考：《南京中医药大学》2016年博士论文

【摘要】：目的：温病“透泄热郁法”是指针对温病“热郁”这一病机所采取的一种透法和泄法相结合的复合型治法。该法以祛邪为目的,针对温病过程中温邪和气郁相互搏结难解的这一病理变化,采取透和泄相结合的综合性治疗。然而,这一温病的特色治法,古今中医文献中仅有些散在的记载,对其认识尚有待客观化和规范化。在中医药界当今注重中医传统研究方法与先进科研技术手段相结合的大背景下,有必要用传统研究方法与现代科学技术相结合,加强对这一温病特色治法的客观化和规范化的研究。本课题沿着这一思路,分别从理论研究、数据挖掘和实验研究三个部分对温病透泄热郁法进行研究。方法：本课题第一部分为理论研究。系统梳理温病透泄热郁法的概念、立法依据、适应证、在温病中的具体应用、古代医家对本法的认识以及这一治法在肺部感染性疾病中的运用等方面。第二部分采取反向挖掘方法。以曹洪欣主编的《温病大成》中记载的明末至民国(公元1642-1949年)120部温病医籍为研究样本,获得温病透泄热郁法的临床运用信息,利用Medcase V3.2数据挖掘系统对其进行建库、挖掘和分析。通过对透泄热郁法在温病治疗中的症状、方药等项集的挖掘,以便分析和研究温病透泄热郁法的方证关系,为临床在温病范围准确应用透泄热郁法治疗温病提供客观依据和方法学参考。第三部分为实验研究。用不同浓度LPS((O、2、10μg/mL)诱导大鼠肺泡巨噬细胞(NR8383),采用Real-time PCR和Western blotting分别检测TLR4的mRNA和蛋白表达水平,Real-time PCR检测细胞因子TNF-α、A20、IL-6和IL-1β的mRNA表达水平,ELISA检测细胞上清液中TNF-α、IL-6和IL-1β的含量。应用NO含量检测试剂盒、ELISA和Real-time PCR,检测经升降散处理的LPS成功诱导的NR8383细胞中主要促炎因子和细胞因子N0、PGE2、iNOS、COX-2、TNF-α、IL-6和IL-1β表达水平；Western blotting分别检测上述细胞浆、核内的p65和IκB-α表达水平。提取细胞总蛋白,Western blotting检测细胞中ERK和p-ERK、JNK、p-JNK、p38和p-p38表达水平。结果：1.理论研究方面。通过古今文献研究发现,所谓温病“透泄热郁法”,是针对温病“热郁”这一病机所采取的一种透法和泄法相结合的复合型治法。其中,透法重在使邪气由深至浅外出而解,泄法重在疏散、排泄邪气,给邪气外透扫清障碍、铺平道路。二者有机配合,能快速而有效地使热和郁相互分离、病邪外透而解。古代医籍中有关温病透泄热郁法的记载零星、散在,未形成规范化和客观化。现代有学者通过临床回顾性分析,肺部感染性疾病“热郁”相关证候呈现出发病率高、致死率高和耐药性强这“三高”的特点,已有学者将温病透泄热郁方应用到肺部感染性疾病的治疗之中,并取得了良好的临床疗效。如曹洪欣等人对2003年北京、广州、天津等地的SARS治疗方案的疗效进行回顾性分析,结果显示,综合类方剂配伍应用统计中,升降散位列第一。上述内容提示,温病透泄热郁法具有一定的理论基础和现代临床研究价值。2.温病医案应用透泄热郁法临床经验要素数据挖掘分析方面检索获得温病透泄热郁法相关温病类医籍21部、医案181例,涉及症状147条,脉象15类,舌象51条,药物186种,症状内关联产生规则30条,舌象内关联产生规则30条,脉象内关联规则30条,用药内关联产生规则94条。通过数据挖掘分析符合纳入标准的181例温病透泄热郁法医案数据,归纳出临床上运用透泄热郁法治疗温病的辨证依据,包括以下几点：常见症：矛盾热象(如发热、目赤、口渴、小便短赤、发斑等,但面色苍白或晦滞无华、无汗、或者少汗、或者但头汗出、四肢不温等),神志异常(烦躁、神昏谵语等),气郁征象(胸闷、气喘、脘痞等)。可见症：恶寒、寒战、身痛、头痛、口唇干裂、咳嗽、咳痰、咽喉肿痛、食欲不振、呕吐、恶心、恶梦纷纭、失眠、大便秘结、便溏、倦怠乏力、发疹,舌质红或红绛,苔黄燥或黄腻,脉沉数。温病透泄热郁法透泄药物配伍模式研究方面：①具体遣方用药时,泄类药物在药物种类和使用频次上,所占比重均较高；透类药物在性味上有一个共性,即皆具有辛味或性味芳香。②温病透泄热郁方并不是机械地由透类药物和泄类药物相加组成,其中还包括其它类药物。③透泄共用的常见药物：淡豆豉+栀子、犀角+连翘、连翘+生地黄、连翘+玄参。3.实验研究方面实验一：通过建立HPLC法同时测定升降散中大黄酸、大黄素、大黄酚中的含量,评估以及证实了组成方剂药材的稳定性和可靠性,这为后续实验的进一步开展,提供了质量监控方面的参考。实验二：LPS能成功诱导NR8383细胞中NF-κB上调,在蛋白表达和m-RNA表达水平均证实,LPS能梯度诱导TLR4高表达,且与LPS浓度呈正相关；同时,LPS能诱导TLR4下游NF-κB的激活,增加下游靶基因编码细胞因子的合成和分泌,且与LPS浓度呈正相关。浓度为2μg/mL、5μg/mL的升降散,能够显著抑制LPS诱导的NR8383细胞中iNOS和COX-2的蛋白表达水平,其下游产物NO、 PGE2、TNF-α、IL-6和IL-1β的表达也随之下降。同时,升降散作用LPS诱导的NR8383细胞后,能够通过抑制p65进入细胞核内以及IκB-α的磷酸化来抑制NF-κB信号通路的激活。
[Abstract]:Objective: the method of febrile disease "transmenting heat depression" is a combined treatment method which is adopted by the pointer to the febrile disease "hot depression". This method is aimed at removing pathogenic factors and aiming at the pathological changes of temperature evil and qi depression in the process of febrile disease. It is necessary to combine traditional methods of traditional Chinese medicine with advanced scientific research and technology in the context of combining traditional Chinese medicine research methods and advanced scientific and technological means. The objective and standardized research of the rule of law. This subject follows this idea to study the febrile diseases through three parts: theoretical research, data mining and experimental research. Method: the first part of this subject is theoretical research. Application, the understanding of this method by ancient doctors and the application of this method in the infectious diseases of the lung. The second part adopts the reverse mining method. The clinical application letter of the method of heat depression and heat depression is obtained with the sample of 120 febrile diseases recorded in the late Ming to the Republic of China (1642-1949 AD), which is the editor of Cao Hongxin. By using the Medcase V3.2 data mining system to build the database, excavate and analyze it. Through the excavation of the symptoms, prescription and other items in the treatment of warm disease in the treatment of febrile diseases in order to analyze and study the relationship between the prescription of heat depression and the method of heat depression in warm disease, so as to provide an objective basis and prescription for the treatment of febrile diseases in the correct range of temperature disease. The third part was the experimental study. The rat alveolar macrophages (NR8383) were induced with different concentrations of LPS (O, 2,10 mu g/mL). The mRNA and protein expression levels of TLR4 were detected by Real-time PCR and Western blotting respectively. The content of TNF- alpha, IL-6 and IL-1 beta. NO content detection kit, ELISA and Real-time PCR were used to detect the main pro-inflammatory factors and cytokines N0, PGE2, iNOS, alpha, and beta expression levels in the NR8383 cells successfully induced by LPS. Expression level. Extract the total cell protein and Western blotting to detect the expression level of ERK and p-ERK, JNK, p-JNK, p38 and p-p38 in cells. Results: 1. theoretical research. Through the study of ancient and modern literature, it is found that the so-called febrile disease "heat release heat depression" is a combination of a combination of penetration and vent methods adopted for the febrile disease "hot depression". Among them, the method is to solve the evil spirit from deep to shallow, and the method is to evacuate, discharge evil, clear the obstacles, and smooth the road. The two organic cooperation can quickly and effectively make the heat and depression separate from each other, and get rid of the disease. Through the clinical retrospective analysis, a clinical retrospective analysis shows that the syndrome of "hot depression" of the pulmonary infection disease is characterized by high incidence, high mortality and high resistance to drug resistance. Some scholars have applied the febrile disease with febrile disease to the treatment of pulmonary infectious diseases and achieved good clinical effect. A retrospective analysis of the curative effect of the SARS treatment program in Beijing, Guangzhou, Tianjin and other places in 2003 was reviewed. The results showed that in the statistics of the combination of comprehensive prescriptions, the number of elevating and dispersing was the first. The above content suggested that the method of febrile disease permeation heat depression has a certain theoretical basis and modern clinical research value for the application of the medical case of.2. febrile disease. The clinical experience factor data mining analysis of the heat depression method retrieves 21 medical records of febrile diseases related to febrile disease, 181 medical cases, including 147 symptoms, 15 pulse images, 51 tongue images, 186 drugs, 30 rules in symptoms, 30 rules in the association of tongue images, 30 association rules in the pulse image, and association rules within the medication. The 94. Through data mining analysis to meet the inclusion criteria of 181 cases of febrile disease, heat depression forensic case data, summed up the clinical use of heat depression to treat febrile diseases, including the following points: common symptoms: fever, such as fever, eye red, thirst, short urine, hair spots and so on, but the face pale or obscure, no Hua, no sweat, or Less sweat, or the sweat out of the head, limbs not warm, etc), abnormal spirit (irritability, deity delirium, etc.), qi depression signs (chest tightness, asthma, epigastric, etc.). Visible symptoms: malaise, shiver, body pain, headache, lip dry crack, cough, phlegm, sore throat, nausea, nightmare, insomnia, stool, loose stool, loose stool, rash, red tongue or red tongue Red crimson, yellowing dry or yellow greasy, pulse sink number. Study on the compatibility mode of leakage of heat depression in febrile disease: (1) the specific proportion of the drug is higher in the type and frequency of drug use when the prescription is used, and the drug has a common character on the sexual taste. It is composed of other drugs and other drugs. (3) the common drugs that are shared by the drugs. (3) the common drugs that are shared by the spillway: light Douchi + gardenia, rhinoceros horn + forsythia, forsythia Forsythia + Dihuang, forsythia forsythi +.3.. The content of rhein, emodin and chrysophanol in lifting and dispersing is determined by establishing HPLC method The stability and reliability of the ingredients were estimated and confirmed, which provided a reference for quality monitoring for further experiments. Experiment two: LPS could successfully induce the up regulation of NF- kappa B in NR8383 cells. Both in protein expression and m-RNA expression level, LPS can induce high expression of TLR4, and it is positively related to the concentration of LPS. At the same time, LPS can induce the activation of NF- kappa B downstream of TLR4, increase the synthesis and secretion of the downstream target gene encoding cytokine, and have a positive correlation with the concentration of LPS. The concentration is 2 mu g/mL, and the rise and drop of 5 mu g/mL can significantly inhibit the protein expression level of iNOS and COX-2 in the NR8383 cells induced by LPS. Meanwhile, the expression of NR8383 cells induced by LPS can inhibit the activation of NF- kappa B signaling pathway by inhibiting the phosphorylation of p65 into the nucleus and I kappa B- alpha.
【学位授予单位】：南京中医药大学
【学位级别】：博士
【学位授予年份】：2016
【分类号】：R254.2

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