本文選題：肺纖維化　切入點(diǎn)：咳嗽　出處：《北京中醫(yī)藥大學(xué)》2014年碩士論文　論文類型：學(xué)位論文

【摘要】：特發(fā)性肺纖維化(idiopathic pulmonary fibrosis,IPF)以進(jìn)行性呼吸困難,隨病情發(fā)展,氣不相續(xù)發(fā)為喘,同時(shí)伴刺激性干咳為主要癥狀。系統(tǒng)整理發(fā)掘仲景辨治咳嗽與喘證的用藥規(guī)律,并結(jié)合近年中醫(yī)藥治療IPF的用藥經(jīng)驗(yàn),制定出了用于治療IPF配伍優(yōu)化的復(fù)方。運(yùn)用混料均勻設(shè)計(jì)的方法將復(fù)方設(shè)計(jì)成藥物組成相同而劑量配比不同的多首復(fù)方,將各復(fù)方作用于以博萊霉素誘導(dǎo)的雄性ICR小鼠肺纖維化模型,以模型小鼠7d、28d死亡率及Hyp含量為藥效指標(biāo)來尋找該復(fù)方最優(yōu)配伍模式,為中醫(yī)臨床治療IPF合理用藥提供實(shí)驗(yàn)依據(jù)。 理論研究 理論研究一：仲景關(guān)于咳嗽與喘證的辨治規(guī)律探析 系統(tǒng)整理并研究《傷寒論》及《金匱要略》中出現(xiàn)的有關(guān)“咳”、“喘”及“短氣”的條文,揭示仲景在治療咳喘方面的用藥規(guī)律。在辨證論治的基礎(chǔ)上,基于藥證相應(yīng)、舍性取用理論,指出仲景治咳常用藥物組合為干姜、細(xì)辛、五味子；對(duì)于喘證的治療仲景擅用麻黃,因麻黃功擅宣肺平喘,為治喘之要藥,且麻黃常與半夏為伍,共奏降逆平喘之功。 理論研究二：用治IPF復(fù)方的選藥依據(jù) 基于中醫(yī)對(duì)IPF病程一般發(fā)展規(guī)律的認(rèn)識(shí),以癥狀學(xué)為切入點(diǎn),認(rèn)為本病早期可歸屬中醫(yī)“咳嗽”與“喘證”的范疇,其基本的病理改變?yōu)榉螝怅巸商摚浑S著病情的發(fā)展,癥狀又可見唇指發(fā)紺、舌質(zhì)紫暗等瘀血證候,以及手足逆冷、皮膚發(fā)涼等陽虛證候；病情進(jìn)一步發(fā)展,又會(huì)出現(xiàn)咳喘加劇、動(dòng)則喘甚等肺腎兩虛證候,并伴見乏力、厭食、消瘦等虛勞證候。基于對(duì)IPF中醫(yī)證候演變發(fā)展規(guī)律的認(rèn)識(shí),從仲景辨治咳喘的規(guī)律、中醫(yī)臨床治療IPF主要方藥的文獻(xiàn)報(bào)道尋找用于治療IPF的中藥復(fù)方,最終確定了用治IPF復(fù)方優(yōu)化篩選的具體藥物組成為：麥冬、法半夏、干姜、五味子、炙麻黃、炙黃芪、黨參、炙甘草、當(dāng)歸、丹參、黃芩、桔梗。 實(shí)驗(yàn)研究 實(shí)驗(yàn)一：用治IPF復(fù)方不同配伍比例的藥效實(shí)驗(yàn)研究 實(shí)驗(yàn)方法： 將195只雄性ICR小鼠隨機(jī)分為13組,即正常組、模型組和11個(gè)中藥復(fù)方組。小鼠麻醉后向氣管內(nèi)注射博萊霉素,造成肺纖維化模型。復(fù)方組小鼠每日給予混料均勻設(shè)計(jì)好的總劑量相等而配比不同的11種復(fù)方,正常組及模型組小鼠給予等量的生理鹽水,連續(xù)灌胃28d。觀察各組小鼠的生存狀態(tài)、體重變化,統(tǒng)計(jì)死亡情況；堿水解法測(cè)定肺組織Hyp含量；HE染色和Masson染色,觀察小鼠肺部病理變化。 實(shí)驗(yàn)結(jié)果及討論： 1小鼠一般狀況變化：第7d時(shí),復(fù)方第1、5組小鼠體重較模型組增加,復(fù)方第11組小鼠體重較模型組下降；第14d時(shí),復(fù)方第1組小鼠體重較模型組增加；第28d時(shí),復(fù)方第1組小鼠體重較模型組增加。 2各復(fù)方對(duì)小鼠死亡率的影響及配伍優(yōu)化：(1)復(fù)方對(duì)小鼠死亡率的影響：第2、5組復(fù)方對(duì)于提高小鼠肺纖維化早期(炎癥期)的生存率有幫助；第1、2、5、6組復(fù)方對(duì)于提高肺纖維化模型小鼠整個(gè)周期的生存率有幫助。(2)各組小鼠28d生存率與28d平均體重有直線相關(guān)關(guān)系,說明肺纖維化小鼠體重越高,生存優(yōu)勢(shì)越明顯。(3)以7d死亡率為藥效指標(biāo)的配伍優(yōu)化：麥冬、當(dāng)歸、五味子、桔梗、黃芩對(duì)于死亡率的降低起主導(dǎo)作用,干姜、炙黃芪對(duì)于死亡率的降低起輔助作用。(4)以28d死亡率為藥效指標(biāo)的配伍優(yōu)化：麥冬、炙黃芪、當(dāng)歸、桔梗四味藥物起主導(dǎo)作用。 3各復(fù)方對(duì)小鼠肺組織28dHyp含量的影響及配伍優(yōu)化：(1)復(fù)方對(duì)小鼠肺組織28dHyp含量的影響：第1組復(fù)方能顯著降低小鼠肺組織Hyp含量,第11組復(fù)方可升高小鼠肺組織Hyp含量。(2)各組小鼠28d死亡率與28dHyp含量有直線相關(guān)關(guān)系,說明有效抑制肺纖維化組織中膠原的沉積對(duì)于生存率的提高有積極意義。(3)以小鼠28d肺組織Hyp含量為藥效指標(biāo)的配伍優(yōu)化：五味子桔梗、炙黃芪、炙甘草、黃芩起主導(dǎo)作用,麥門冬、丹參起一定的輔助作用。 4肺組織形態(tài)學(xué)觀察：(1)HE染色觀察炎癥細(xì)胞浸潤：復(fù)方1、2、5組在一定程度上有減輕小鼠肺泡炎,促進(jìn)肺實(shí)質(zhì)細(xì)胞修復(fù)的作用,并具有一定的抑制肺纖維化形成的作用。復(fù)方3、6組在一定程度上具有減輕小鼠肺泡炎和促進(jìn)肺實(shí)質(zhì)修復(fù)的作用。(2) Masson染色觀察纖維化程度：復(fù)方第1、2、5、6組膠原含量較模型組明顯減少,表明這些復(fù)方在減輕膠原纖維沉積和抗纖維化方面的作用顯著。 以Hyp含量為藥效指標(biāo)的優(yōu)化出的藥物基本包含了以7d死亡率和28d死亡率為藥效指標(biāo)優(yōu)化出的藥物。說明優(yōu)化結(jié)果因藥效指標(biāo)的不同而有所差異,但無論以哪個(gè)指標(biāo)進(jìn)行優(yōu)化,結(jié)果中的主要藥物均在五味子、桔梗、炙黃芪、炙甘草、黃芩、麥門冬、當(dāng)歸之中,只是它們的配伍比例有所不同,提示這些藥物配伍組成的復(fù)方對(duì)于肺纖維化的治療有一定作用。 實(shí)驗(yàn)二：藥效學(xué)再驗(yàn)證實(shí)驗(yàn) 實(shí)驗(yàn)方法： 將60只雄性ICR小鼠隨機(jī)分為6組,分別為正常組、模型組、復(fù)方第5組、7d死亡率優(yōu)化組、28d死亡率優(yōu)化組、序貫治療組。檢測(cè)各組小鼠肺組織Hyp含量及CTGF、IL-6、Foxp3mRNA含量。 實(shí)驗(yàn)結(jié)果及討論： 1序貫治療組無論對(duì)于減少肺組織中膠原的沉積,還是下調(diào)某些細(xì)胞因子(如CTGF、IL-6)的基因表達(dá)水平都有顯著療效,這為IPF的分期治療(早期在補(bǔ)肺滋陰的同時(shí)兼顧清熱解毒,后期注重滋陰助陽、補(bǔ)氣活血養(yǎng)血)提供了實(shí)驗(yàn)依據(jù)。 2序貫治療組和28d死亡率優(yōu)化組復(fù)方可升高Foxp3mRNA表達(dá)水平,28d死亡率優(yōu)化組復(fù)方可降低CTGFmRNA和IL-6mRNA表達(dá)水平,序貫治療組復(fù)方可降低CTGFmRNA表達(dá)水平。結(jié)果表明CTGF及IL-6基因表達(dá)過量、Foxp3基因表達(dá)不足可能是IPF發(fā)病的重要因素。
[Abstract]:Idiopathic pulmonary fibrosis (idiopathic pulmonary, fibrosis, IPF) with dyspnea, with the progression of the disease, not continue to breathe air, at the same time with the irritating cough as the main symptom. The system finishing excavation of Zhongjing cough and asthma and drug laws, combined with the clinical experience of Chinese medicine in treating IPF in recent years. Developed for treatment of compound optimization. Using IPF with uniform mixing design method will be designed into the same composition and compound drug dose of different proportion of the first compound, the compound effect on male ICR mice pulmonary fibrosis model induced by bleomycin in mice, with 7d, 28d and Hyp content for the efficacy of mortality to find the optimal parameters of compound compatibility mode, to provide the experimental basis for the clinical treatment of IPF medication.
theoretical research
A study on the theory of Zhongjing on cough and asthma treatment rules of
Collect and Study on typhoid fever and < < > > in the Golden Chamber "cough", "asthma" and "short" provisions, reveal Zhongjing medication rules in the treatment of cough and asthma aspects. In the basis of syndrome differentiation and treatment, based on the corresponding medicine, take care theory, pointed out that the secondary kageharu cough drugs commonly used combination of ginger, asarum, Schisandra chinensis; for asthma treatment in Zhongjing using ephedra, ephedra for work at Xuanfei asthma, for the treatment of asthma to medicine, and Ma Huang and Pinellia company, played a total of relieving asthma of power.
Theoretical study two: the basis for the treatment of IPF compound
Understanding the general law of development of IPF course based on traditional Chinese medicine, with symptoms as the starting point, that the disease attributable TCM "cough" and "asthma" category, the basic pathological changes of the lung qi and yin deficiency; with the development of disease symptoms, but also refers to the lip cyanosis, dark purple tongue blood stasis wait, and hand foot skin against the cold cold Yang deficiency syndrome; further development of the disease, will appear cough aggravating, while activing lung and kidney deficiency syndromes such as two, and accompanied by fatigue, anorexia, weight loss and other asthenia syndrome. Understanding of IPF TCM syndrome evolution law based on the Zhongjing treating cough of TCM clinical treatment of IPF mainly medicine reported in the literature for Chinese herbal compound in the treatment of IPF, and ultimately determine the composition of IPF treated with compound optimization specific drug screening: Ophiopogon japonicus, pinellia, ginger, ephedra, Schisandra, astragalus root, dangshen, roasted sweet Grass, angelica, Salvia miltiorrhiza, Scutellaria, Platycodon.
experimental study
Experiment 1: Experimental Study on the efficacy of different proportions of IPF compound
Experimental methods:
195 male ICR mice were randomly divided into 13 groups: normal group, model group and 11 compound Chinese medicine group. The mice were anesthetized after intratracheal injection of bleomycin, pulmonary fibrosis caused by compound model. Mice were given daily uniform mixing design total dose equal and different ratio of 11 kinds of compound, normal saline group and model group were given equal weight change, continuous intragastric 28d. mice were observed, the survival of the state, death statistics; Hyp content of lung tissue was measured by alkaline hydrolysis; HE staining and Masson staining to observe the pathologic changes in the lung.
Experimental results and discussions:
1, general condition changes of mice: at 7d, the weight of compound 1,5 group increased compared with the model group, and the weight of compound eleventh group mice decreased compared with the model group. At the time of 14d, the weight of compound first group mice increased compared with the model group. When 28d was added, the weight of first groups of compound mice increased compared with the model group.
2 the compound effect on the mortality of mice and the compatibility of Optimization: (1) the effect of compound on the mortality of mice: Group 2,5 compound for improving early pulmonary fibrosis in mice (inflammation) the survival rate of help; the group 1,2,5,6 compound to help improve the survival rate of the entire cycle of the pulmonary fibrosis model in mice. (2) a a linear correlation between the average weight of the mice survival rate of 28d and 28d, the higher the weight of mouse pulmonary fibrosis, a survival advantage is more obvious. (3) to 7d mortality formula optimization efficacy indicators: Radix, Angelica sinensis, Schisandra chinensis, Platycodon grandiflorum, Scutellaria for reducing the mortality play a leading role, ginger, astragalus root to reduce mortality play a supporting role. (4) to 28d mortality formula optimization efficacy indicators: Radix, astragalus root, angelica, Balloonflower four drugs play a leading role.
3 the compound effect on the 28dHyp content in the lung tissue of mice and the compatibility of Optimization: (1) effect on the content of 28dHyp in the lung tissue of mice in first groups: compound can significantly reduce the Hyp content in lung tissue of mice, eleventh groups of compound can increase the content of Hyp in lung tissue of mice. (2) the death rate of mice 28d and 28dHyp content line correlation between that inhibit collagen deposition in pulmonary fibrosis has a positive significance to improve the survival rate. (3) the content of Hyp in lung tissue of mice 28d compatibility optimization efficacy indicators: Schisandra astragalus root, licorice root, Platycodon root, Scutellaria plays a leading role, Ophiopogon japonicus, Salvia miltiorrhiza auxiliary function.
Observation of 4 lung tissue morphology: (1) to observe the infiltration of inflammatory cells in HE staining: 1,2,5 group compound to a certain extent alleviated alveolitis in mice, promote the repair of lung parenchyma cells, and inhibit the formation of pulmonary fibrosis has certain effect. The compound 3,6 group to a certain extent with ease and promote pulmonary alveolitis in mice the essence of repair. (2) to observe the degree of fibrosis Masson staining: group 1,2,5,6 compound collagen content was significantly reduced compared with model group showed that the compound in reducing collagen deposition and anti fibrosis effect.
The contents of Hyp was optimized by drug efficacy index basically contains 7d and 28d mortality for drug efficacy index optimization. The optimization results show that the pharmacodynamic indexes vary, but whichever index is optimized, the main results in drug Schisandra, astragalus root, licorice root, Platycodon grandiflorum. Baicalin, Ophiopogon, angelica, but they have different compatibility proportion, suggesting that compound composition of these drugs have certain effect for the treatment of pulmonary fibrosis.
Experiment two: revalidation experiment of pharmacodynamics
Experimental methods:
60 male ICR mice were randomly divided into 6 groups: normal group, model group, compound fifth group, 7d mortality optimization group, 28d mortality optimization group and sequential treatment group. The Hyp content and CTGF, IL-6 and Foxp3mRNA contents in lung tissue of each group were detected.
Experimental results and discussions:
1 sequential therapy group for reducing collagen deposition in the lung tissue, or down-regulation of some cytokines (CTGF, IL-6) gene expression levels have a significant effect, which is IPF (in the early stage treatment of Bufei Ziyin both detoxification, late on the Yin and Yang, Qi and nourishing blood) provides on the basis of experiment.
2 sequential therapy group and 28d group compound can increase the mortality to optimize the expression level of Foxp3mRNA, 28d group compound optimization can reduce CTGFmRNA mortality and IL-6mRNA expression level, sequential therapy group compound can reduce the expression level of CTGFmRNA. The results showed that overexpression of CTGF and IL-6, the expression of Foxp3 gene deficiency may be an important factor in the pathogenesis of IPF.

【學(xué)位授予單位】：北京中醫(yī)藥大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R289;R259

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 周慶偉,李素云;《金匱要略》皂莢丸治療慢性阻塞性肺病痰濁阻肺型的臨床研究[J];中國醫(yī)藥學(xué)報(bào);1997年04期

2 何彥俠,辛建保;血管生成在肺纖維化發(fā)病機(jī)制中的意義[J];國外醫(yī)學(xué).呼吸系統(tǒng)分冊(cè);2004年S1期

3 王興勝;肺纖維化治療的研究進(jìn)展[J];國外醫(yī)學(xué)(內(nèi)科學(xué)分冊(cè));2001年09期

4 邵長周;細(xì)胞因子在肺纖維化形成中的作用[J];國外醫(yī)學(xué)(內(nèi)科學(xué)分冊(cè));2001年11期

5 童琦燕;;王安康治療特發(fā)性肺間質(zhì)纖維化的經(jīng)驗(yàn)[J];湖北中醫(yī)雜志;2007年12期

6 趙蘭才,武維屏;肺間質(zhì)纖維化的中醫(yī)研究進(jìn)展述評(píng)(綜述)[J];北京中醫(yī)藥大學(xué)學(xué)報(bào);2000年04期

7 韓文銘,何麗娜,黃燕,班俊敏,湯潔;中性粒細(xì)胞與白細(xì)胞介素-8在特發(fā)性肺間質(zhì)纖維化中的作用[J];臨床內(nèi)科雜志;2002年02期

8 張德勝 ,任鳳英;糖皮質(zhì)激素在間質(zhì)性肺病中的應(yīng)用[J];中原醫(yī)刊;2002年09期

9 戴令娟，侯杰，蔡后榮，王偉，孟繁青，倪瑾;肺纖維化動(dòng)物模型肺組織形態(tài)定量分析[J];南京鐵道醫(yī)學(xué)院學(xué)報(bào);1996年01期

10 智屹惠;曹世宏教授論治肺間質(zhì)纖維化[J];南京中醫(yī)藥大學(xué)學(xué)報(bào)(自然科學(xué)版);2001年03期

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