發(fā)布時間：2018-01-10 20:21

  本文關(guān)鍵詞：從腸道菌群角度探討溫通方對過敏性哮喘大鼠的治療作用　出處：《北京中醫(yī)藥大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 腸道菌群結(jié)構(gòu) 過敏性哮喘 溫通方 Th1/Th2 Th17/Treg

【摘要】：目的:通過研究溫通方干預(yù)過敏性哮喘大鼠模型后的一般狀態(tài)、氣道反應(yīng)性、血EOS計數(shù)、肺組織病理形態(tài)學(xué)、腸道菌群結(jié)構(gòu)等指標(biāo)的變化情況,探索過敏性哮喘大鼠模型是否存在腸道菌群失衡、溫通方能否通過影響腸道菌群干預(yù)過敏性哮喘。方法:將40只大鼠隨機(jī)分為4組(空白對照組,模型對照組,地塞米松組,溫通方組),適應(yīng)性喂養(yǎng)一周,造模組以O(shè)VA/Al(OH)3(1mgOVA+30mgAl(OH)3凝膠)于1ml 0.9%生理鹽水中五點注射,同時0.2ml百白破疫苗腹腔注射(第1天和第8天,共2次)致敏大鼠,后用1%OVA生理鹽水霧化吸入激發(fā)(第15天開始用,每天一次,20min/次,共激發(fā)7次)復(fù)制過敏性哮喘大鼠模型,造模結(jié)束后予每日灌胃一次,共14天。溫通方組灌胃量為1.34g/kg的水提液浸膏;地塞米松組灌胃量為0.81mg/kg的地塞米松片劑;其他組灌胃使用等量生理鹽水。實驗過程中觀察大鼠一般情況,灌胃結(jié)束后測定大鼠氣道反應(yīng)性(呼氣阻力),取外周血測定血EOS計數(shù),取大鼠肺組織進(jìn)行HE染色分析病理損傷程度,取盲腸內(nèi)容物進(jìn)行DNA提取、擴(kuò)增、16SrDNA測序分析,觀察腸道菌群結(jié)構(gòu)的變化。結(jié)果:1.造模結(jié)束后除空白對照組大鼠外,其余各組大鼠均出現(xiàn)不同程度的噴嚏、焦躁易動、撓鼻、呼吸頻促等典型哮喘癥狀。灌胃結(jié)束后,模型對照組哮喘癥狀最為顯著,與模型對照組相比,地塞米松組、溫通方組癥狀顯著減輕。2.肺組織HE染色:空白對照組未見炎性細(xì)胞浸潤,且肺泡、支氣管粘膜上皮結(jié)構(gòu)完整清晰,支氣管無痙攣;模型對照組、地塞米松組及溫通方組均可見肺組織炎性細(xì)胞浸潤,支氣管黏膜上皮細(xì)胞排列紊亂、成片脫落,肺泡間隔增厚,支氣管痙攣等典型支氣管哮喘病理表現(xiàn),但病理損害輕重程度不同,病變程度從高到低依次為模型對照組、溫通方組,地塞米松組、空白對照組。3.氣道反應(yīng)性測定:模型對照組基礎(chǔ)呼氣阻力、各Ach激發(fā)濃度下呼氣阻力與空白對照組相比差異有統(tǒng)計學(xué)意義(P0.05),溫通方組和地塞米松組在Ach激發(fā)濃度為50、100ug/kg時呼氣阻力較模型對照組均有改善,差異具有統(tǒng)計學(xué)意義(P0.05);在各Ach激發(fā)濃度下,溫通方組和地塞米松組呼氣阻力測定結(jié)果差異無統(tǒng)計學(xué)意義(P0.05)。4.血EOS計數(shù):與空白對照組相比,模型對照組、溫通方組、地塞米松組與其差異具有顯著統(tǒng)計學(xué)意義,(P0.01)與模型對照組相比,地塞米松組、溫通方組與其差異具有顯著統(tǒng)計學(xué)意義(P0.01)與地塞米松組相比,溫通方組與其差異具有顯著統(tǒng)計學(xué)意義(P0.01)。5.腸道菌群結(jié)構(gòu):α多樣性:Chao1指數(shù)顯示,與模型對照組相比,空白對照組、地塞米松組、溫通方組與其差異有統(tǒng)計學(xué)意義(P0.05);Shannon指數(shù)和Simpson指數(shù)顯示各組差異無統(tǒng)計學(xué)意義(P0.05)。β多樣性:地塞米松組腸道菌群結(jié)構(gòu)與其余各組差異最大,溫通方組菌群結(jié)構(gòu)最接近空白對照組大鼠。菌群結(jié)構(gòu)差異分析:與空白對照組相比,模型對照組 Akkermansia、Oscillospira、Coprococcus、Phascolarctobacterium、Roseburia、Prevotella 豐度增加(Ratiokb/mx0.5),Dorea 豐度降低(Ratio kb/mx2);與模型對照組相比,地塞米松組Phascolarctobacterium、Prevotella、Dorea 豐度增加(Ratio dex/mx2),Akkermansia、Oscillospira 豐度降低(Ratio dex/mx0.5);與模型對照組相比,溫通方組Coprococcus、Phascolarctobacterium 豐度增加(Ratio zy/mx2),Akkermansia、Oscillospira、Roseburia、Prevotella豐度降低(Ratio zy/mx0.5);與空白對照組相比,地塞米松組Akkermansia、Coprococcus、Phascolarctobacterium、Prevotella 豐度增加(Ratio dex/kb2),Lactobacillus豐度降低(Ratiodex/kb0.5);與空白對照組相比,溫通方組 Coprococcus、Phascolarctobacterium 豐度增加(Ratiozy/kb2),其余菌屬與空白對照組無顯著差異(0.5Ratiozy/kb2);與地塞米松組相比,溫通方組Coprococcus豐度增加(Ratio dex/zy2),Akkermansia、Phascolarctobacterium、Roseburia、Prevotella、Dorea 豐度降低(Ratio dex/zy0.5)。結(jié)論:1.溫通方對過敏性哮喘大鼠的一般狀況、氣道反應(yīng)性、血EOS計數(shù)、肺組織病理產(chǎn)生了有利作用,說明本方具有緩解過敏性哮喘大鼠癥狀、降低氣道反應(yīng)性、減輕氣道炎癥的效應(yīng)。2.過敏性哮喘大鼠模型存在腸道菌群紊亂。治療后地塞米松組大鼠腸道菌群差異較空白對照組較大,提示其對過敏性哮喘的治療作用不能從調(diào)整腸道菌群結(jié)構(gòu)解釋;治療后的溫通方組大鼠模型在癥狀、指標(biāo)緩解的同時,腸道菌群結(jié)構(gòu)亦發(fā)生了一定改變,較地塞米松組更接近于空白對照組,表明中藥湯劑溫通方可能在一定程度上通過影響腸道菌群結(jié)構(gòu)緩解了過敏性哮喘的癥狀和相關(guān)指標(biāo)。
[Abstract]:Objective: general state intervention after the rat model of allergic asthma through the study of the temperature Tongfang, airway reactivity, blood EOS count, lung tissue pathological changes of the intestinal flora structure, etc, to explore the rat model of allergic asthma is the presence of intestinal flora imbalance, temperature Tongfang can pass through the intestinal flora intervention allergic asthma. Methods: 40 rats were randomly divided into 4 groups (blank control group, model control group, dexamethasone group, temperature Tongfang group), fed for one week, the model with OVA/Al (OH) 3 (1mgOVA+30mgAl (OH) 3 gel) in five 1ml 0.9% normal saline injection at the same time, 0.2ml intraperitoneal injection of DPT vaccine (first days and eighth days, a total of 2 times) in sensitized rats, with 1%OVA saline aerosol inhalation (start with, once every fifteenth days, 20min/ times a day, a total of 7 excited) copy the rat model of allergic asthma, after the modeling for daily irrigation A stomach, a total of 14 days. The temperature Tongfang group gavage volume extract 1.34g/kg water; dexamethasone group gavage for dexamethasone tablets 0.81mg/kg; the other group by gavage using saline. Observe the general condition of the rats in the experimental process, the determination of airway reactivity in rats after gavage (expiratory resistance), taking peripheral blood serum EOS count was performed with rat lung tissue HE staining analysis of pathological injury degree, cecal contents were collected for DNA extraction, 16SrDNA amplification, sequencing analysis, observe the changes of intestinal microflora. Results: 1. after the modeling except blank control group rats and other groups the rats showed varying degrees of anxiety, sneezing, nose, breathing frequency and promoting typical asthma symptoms. After gavage, model control group, asthma symptoms were the most significant, compared with model control group, dexamethasone group, Tongfang group significantly reduce the temperature and symptoms of.2. in lung tissue HE staining Color: blank control group, no inflammatory cell infiltration, alveolar and bronchial mucosa, epithelial structure clear, no bronchial spasm; model group, dexamethasone group and temperature Tongfang were found in lung tissue inflammatory cell infiltration, bronchial epithelial cells arranged in disorder, a loss, alveolar septal thickening, bronchial bronchial spasm the pathology of asthma, but the pathological damage degree, lesion degree from high to low was model group, temperature Tongfang group, dexamethasone group,.3. control group determination of airway reactivity: blank model control group based Ach expiratory resistance, stimulate concentration of expiratory resistance compared with the control group had significant difference (P0.05), temperature Tongfang group and dexamethasone group at a concentration of 50100ug/kg of expiratory resistance than the model control group were improved at Ach excitation, the difference was statistically significant (P0.05); in the Ach Stimulate concentration, temperature Tongfang group and dexamethasone group expiratory resistance determination results were not statistically significant (P0.05).4. blood EOS count: compared with the control group, model control group, temperature Tongfang group, dexamethasone group and there was a statistically significant difference (P0.01) compared with model control group, dexamethasone group, was statistically significant the temperature difference and its significance Tongfang group (P0.01) compared with the dexamethasone group, there was a statistically significant difference in temperature Tongfang group and (P0.01).5. intestinal flora: alpha diversity: Chao1 index shows that, compared with the model control group, blank control group, dexamethasone group, was statistically significant difference with the temperature Tongfang group (P0.05); Shannon index and Simpson index showed no significant difference between groups (P0.05). Beta diversity: dexamethasone group, intestinal microflora and the rest of the maximum temperature difference, Tongfang group flora structure Close to the blank control group rats. Analysis of microbial community structure difference: compared with the control group, model control group, Akkermansia, Oscillospira, Coprococcus, Phascolarctobacterium, Roseburia, Prevotella, Dorea (Ratiokb/mx0.5) increased abundance abundance decreased (Ratio kb/mx2); compared with the model group, dexamethasone group, Phascolarctobacterium, Prevotella, increased Dorea abundance (Ratio dex/mx2, Akkermansia, Oscillospira) (Ratio dex/mx0.5); abundance decreased compared with the model group, Coprococcus group Tongfang temperature, increased Phascolarctobacterium abundance (Ratio zy/mx2), Akkermansia, Oscillospira, Roseburia, Prevotella (Ratio zy/mx0.5); abundance decreased compared with the control group, dexamethasone group, Akkermansia, Coprococcus, Phascolarctobacterium, Prevotella abundance increased (Ratio dex/kb2), Lactobacillus (Ratiodex/kb0.5); and reduce the abundance of blank According to the temperature Tongfang group, group Coprococcus, Phascolarctobacterium (Ratiozy/kb2), the increase of the abundance of bacteria had no significant difference with the control group (0.5Ratiozy/kb2); compared with the dexamethasone group, temperature Tongfang group increased Coprococcus abundance (Ratio dex/zy2), Akkermansia, Phascolarctobacterium, Roseburia, Prevotella, Dorea (Ratio dex/zy0.5) abundance decreased. Conclusion: the general condition of 1. temperature Tongfang on rats with allergic asthma, airway reactivity, blood EOS count, lung pathology had a beneficial effect, the party can relieve rat allergic asthma symptoms, airway reactivity, intestinal flora disturbance effect.2. rat model of allergic asthma, reduce airway inflammation. After treatment the difference of intestinal flora of rats in the dexamethasone group group than the control group, suggesting that the effect on the treatment of allergic asthma is not from the adjustment of the intestinal flora structure interpretation The temperature Tongfang after treatment; the rats in the model group in symptom remission index at the same time, the structure of intestinal flora has also undergone some changes, compared with the dexamethasone group close to the blank control group, showed that the decoction temperature Tongfang may to a certain extent affected by the structure of intestinal flora to ease the symptoms of allergic asthma and related indicators.

【學(xué)位授予單位】：北京中醫(yī)藥大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R285.5

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 趙凡;李春保;;腸道菌Akkermansia muciniphila的特性及其與機(jī)體健康的關(guān)系[J];微生物學(xué)通報;2017年06期

2 劉力維;趙霞;;基于黃芪多糖的藥理學(xué)研究探討其在哮喘緩解期的作用[J];遼寧中醫(yī)雜志;2016年08期

3 王金磊;李承德;孫宏偉;毛淑梅;張曉俊;曲梅花;;黃芪多糖抑制NF-κB/MAPK信號通路和改善哮喘大鼠氣道炎癥的作用[J];中國藥理學(xué)通報;2016年04期

4 鄧鳴;張毅;;固本定喘方治療支氣管哮喘患者臨床觀察[J];中國中醫(yī)急癥;2016年02期

5 楊茜;李曉愚;斯丹;楊致邦;賀中原;張楠晨;張釤釤;石中全;;飲水中地塞米松污染對小鼠腸道菌群的影響[J];南方醫(yī)科大學(xué)學(xué)報;2016年02期

6 凌昊;趙霞;;黃芪多糖治療哮喘的機(jī)制研究進(jìn)展及展望[J];浙江中醫(yī)藥大學(xué)學(xué)報;2016年01期

7 李靜;陳靖;;三拗湯合三子養(yǎng)親湯加減治療過敏性哮喘急性期的臨床觀察[J];中國中醫(yī)急癥;2015年09期

8 丁利忠;李春娟;金東明;王彩霞;;金東明教授應(yīng)用五味子類經(jīng)方治療小兒哮喘經(jīng)驗[J];中國處方藥;2015年06期

9 楊丹紅;蔡明華;陳曉軍;;三伏灸治療過敏性哮喘臨床療效分析[J];浙江中醫(yī)雜志;2014年10期

10 王榮寶;楊翠萍;;麻黃葶藶湯合抗敏煎治療過敏性哮喘40例臨床觀察[J];湖南中醫(yī)雜志;2014年08期

相關(guān)博士學(xué)位論文 前5條

1 孔艷華;基于腸道菌群微生態(tài)失衡的理肺湯干預(yù)慢性阻塞性肺疾病的機(jī)制研究[D];北京中醫(yī)藥大學(xué);2016年

2 孫慧媛;支氣管哮喘慢性持續(xù)期證候特征與“肺脾為核心的臟腑整體辨證”的應(yīng)用研究[D];北京中醫(yī)藥大學(xué);2015年

3 李四維;芪味理肺湯調(diào)控轉(zhuǎn)錄因子活化蛋白-1（AP-1）治療支氣管哮喘的機(jī)制研究[D];北京中醫(yī)藥大學(xué);2014年

4 馮曉凱;我國支氣管哮喘患病情況及相關(guān)危險因素的流行病學(xué)調(diào)查[D];北京協(xié)和醫(yī)學(xué)院;2014年

5 楊君莉;口服Lal對正常新生鼠、成年鼠脾臟Th1/Th2平衡及誘導(dǎo)哮喘模型的影響[D];山東大學(xué);2011年

相關(guān)碩士學(xué)位論文 前4條

1 張妍迪;芪味理肺湯對大鼠支氣管哮喘模型NF-κB、IκBα及細(xì)胞因子分泌的影響[D];北京中醫(yī)藥大學(xué);2014年

2 曹肖t2;黃芪多糖對哮喘小鼠CD_4~+T細(xì)胞作用的研究[D];吉林大學(xué);2014年

3 葉恬吟;基于數(shù)據(jù)挖掘的周仲瑛教授哮喘辨證思路及組方用藥規(guī)律研究[D];南京中醫(yī)藥大學(xué);2010年

4 馬紅玲;雙歧桿菌對過敏性哮喘兒童DC成熟及其分泌細(xì)胞因子的影響[D];中國醫(yī)科大學(xué);2009年

，

本文編號：1406675