本文選題：溫病透泄熱郁法 + 方證理論��； 參考：《南京中醫(yī)藥大學(xué)》2016年博士論文

【摘要】：目的：溫病“透泄熱郁法”是指針對溫病“熱郁”這一病機(jī)所采取的一種透法和泄法相結(jié)合的復(fù)合型治法。該法以祛邪為目的,針對溫病過程中溫邪和氣郁相互搏結(jié)難解的這一病理變化,采取透和泄相結(jié)合的綜合性治療。然而,這一溫病的特色治法,古今中醫(yī)文獻(xiàn)中僅有些散在的記載,對其認(rèn)識尚有待客觀化和規(guī)范化。在中醫(yī)藥界當(dāng)今注重中醫(yī)傳統(tǒng)研究方法與先進(jìn)科研技術(shù)手段相結(jié)合的大背景下,有必要用傳統(tǒng)研究方法與現(xiàn)代科學(xué)技術(shù)相結(jié)合,加強(qiáng)對這一溫病特色治法的客觀化和規(guī)范化的研究。本課題沿著這一思路,分別從理論研究、數(shù)據(jù)挖掘和實驗研究三個部分對溫病透泄熱郁法進(jìn)行研究。方法：本課題第一部分為理論研究。系統(tǒng)梳理溫病透泄熱郁法的概念、立法依據(jù)、適應(yīng)證、在溫病中的具體應(yīng)用、古代醫(yī)家對本法的認(rèn)識以及這一治法在肺部感染性疾病中的運用等方面。第二部分采取反向挖掘方法。以曹洪欣主編的《溫病大成》中記載的明末至民國(公元1642-1949年)120部溫病醫(yī)籍為研究樣本,獲得溫病透泄熱郁法的臨床運用信息,利用Medcase V3.2數(shù)據(jù)挖掘系統(tǒng)對其進(jìn)行建庫、挖掘和分析。通過對透泄熱郁法在溫病治療中的癥狀、方藥等項集的挖掘,以便分析和研究溫病透泄熱郁法的方證關(guān)系,為臨床在溫病范圍準(zhǔn)確應(yīng)用透泄熱郁法治療溫病提供客觀依據(jù)和方法學(xué)參考。第三部分為實驗研究。用不同濃度LPS((O、2、10μg/mL)誘導(dǎo)大鼠肺泡巨噬細(xì)胞(NR8383),采用Real-time PCR和Western blotting分別檢測TLR4的mRNA和蛋白表達(dá)水平,Real-time PCR檢測細(xì)胞因子TNF-α、A20、IL-6和IL-1β的mRNA表達(dá)水平,ELISA檢測細(xì)胞上清液中TNF-α、IL-6和IL-1β的含量。應(yīng)用NO含量檢測試劑盒、ELISA和Real-time PCR,檢測經(jīng)升降散處理的LPS成功誘導(dǎo)的NR8383細(xì)胞中主要促炎因子和細(xì)胞因子N0、PGE2、iNOS、COX-2、TNF-α、IL-6和IL-1β表達(dá)水平；Western blotting分別檢測上述細(xì)胞漿、核內(nèi)的p65和IκB-α表達(dá)水平。提取細(xì)胞總蛋白,Western blotting檢測細(xì)胞中ERK和p-ERK、JNK、p-JNK、p38和p-p38表達(dá)水平。結(jié)果：1.理論研究方面。通過古今文獻(xiàn)研究發(fā)現(xiàn),所謂溫病“透泄熱郁法”,是針對溫病“熱郁”這一病機(jī)所采取的一種透法和泄法相結(jié)合的復(fù)合型治法。其中,透法重在使邪氣由深至淺外出而解,泄法重在疏散、排泄邪氣,給邪氣外透掃清障礙、鋪平道路。二者有機(jī)配合,能快速而有效地使熱和郁相互分離、病邪外透而解。古代醫(yī)籍中有關(guān)溫病透泄熱郁法的記載零星、散在,未形成規(guī)范化和客觀化�，F(xiàn)代有學(xué)者通過臨床回顧性分析,肺部感染性疾病“熱郁”相關(guān)證候呈現(xiàn)出發(fā)病率高、致死率高和耐藥性強(qiáng)這“三高”的特點,已有學(xué)者將溫病透泄熱郁方應(yīng)用到肺部感染性疾病的治療之中,并取得了良好的臨床療效。如曹洪欣等人對2003年北京、廣州、天津等地的SARS治療方案的療效進(jìn)行回顧性分析,結(jié)果顯示,綜合類方劑配伍應(yīng)用統(tǒng)計中,升降散位列第一。上述內(nèi)容提示,溫病透泄熱郁法具有一定的理論基礎(chǔ)和現(xiàn)代臨床研究價值。2.溫病醫(yī)案應(yīng)用透泄熱郁法臨床經(jīng)驗要素數(shù)據(jù)挖掘分析方面檢索獲得溫病透泄熱郁法相關(guān)溫病類醫(yī)籍21部、醫(yī)案181例,涉及癥狀147條,脈象15類,舌象51條,藥物186種,癥狀內(nèi)關(guān)聯(lián)產(chǎn)生規(guī)則30條,舌象內(nèi)關(guān)聯(lián)產(chǎn)生規(guī)則30條,脈象內(nèi)關(guān)聯(lián)規(guī)則30條,用藥內(nèi)關(guān)聯(lián)產(chǎn)生規(guī)則94條。通過數(shù)據(jù)挖掘分析符合納入標(biāo)準(zhǔn)的181例溫病透泄熱郁法醫(yī)案數(shù)據(jù),歸納出臨床上運用透泄熱郁法治療溫病的辨證依據(jù),包括以下幾點：常見癥：矛盾熱象(如發(fā)熱、目赤、口渴、小便短赤、發(fā)斑等,但面色蒼白或晦滯無華、無汗、或者少汗、或者但頭汗出、四肢不溫等),神志異常(煩躁、神昏譫語等),氣郁征象(胸悶、氣喘、脘痞等)。可見癥：惡寒、寒戰(zhàn)、身痛、頭痛、口唇干裂、咳嗽、咳痰、咽喉腫痛、食欲不振、嘔吐、惡心、惡夢紛紜、失眠、大便秘結(jié)、便溏、倦怠乏力、發(fā)疹,舌質(zhì)紅或紅絳,苔黃燥或黃膩,脈沉數(shù)。溫病透泄熱郁法透泄藥物配伍模式研究方面：①具體遣方用藥時,泄類藥物在藥物種類和使用頻次上,所占比重均較高；透類藥物在性味上有一個共性,即皆具有辛味或性味芳香。②溫病透泄熱郁方并不是機(jī)械地由透類藥物和泄類藥物相加組成,其中還包括其它類藥物。③透泄共用的常見藥物：淡豆豉+梔子、犀角+連翹、連翹+生地黃、連翹+玄參。3.實驗研究方面實驗一：通過建立HPLC法同時測定升降散中大黃酸、大黃素、大黃酚中的含量,評估以及證實了組成方劑藥材的穩(wěn)定性和可靠性,這為后續(xù)實驗的進(jìn)一步開展,提供了質(zhì)量監(jiān)控方面的參考。實驗二：LPS能成功誘導(dǎo)NR8383細(xì)胞中NF-κB上調(diào),在蛋白表達(dá)和m-RNA表達(dá)水平均證實,LPS能梯度誘導(dǎo)TLR4高表達(dá),且與LPS濃度呈正相關(guān)；同時,LPS能誘導(dǎo)TLR4下游NF-κB的激活,增加下游靶基因編碼細(xì)胞因子的合成和分泌,且與LPS濃度呈正相關(guān)。濃度為2μg/mL、5μg/mL的升降散,能夠顯著抑制LPS誘導(dǎo)的NR8383細(xì)胞中iNOS和COX-2的蛋白表達(dá)水平,其下游產(chǎn)物NO、 PGE2、TNF-α、IL-6和IL-1β的表達(dá)也隨之下降。同時,升降散作用LPS誘導(dǎo)的NR8383細(xì)胞后,能夠通過抑制p65進(jìn)入細(xì)胞核內(nèi)以及IκB-α的磷酸化來抑制NF-κB信號通路的激活。
[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.
【學(xué)位授予單位】：南京中醫(yī)藥大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2016
【分類號】：R254.2

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