本文選題：支氣管哮喘 + 穴位注射�。� 參考：《河北醫(yī)科大學》2017年碩士論文

【摘要】：目的:觀察穴位注射草分枝桿菌與中藥穴位貼敷對支氣管哮喘大鼠血清免疫球蛋白E(Ig E)水平、肺組織白介素10(IL-10)、白介素17(IL-17)、細胞粘附因子1(ICAM-1)表達的影響,探討穴位注射、穴位貼敷中醫(yī)外治法對支氣管哮喘的作用機制,為中醫(yī)外治法治療支氣管哮喘的臨床應用提供有力依據及參考。方法:SPF級Wistar雄性大鼠60只適應性喂養(yǎng)一周,隨機分為對照組、模型組、草分枝桿菌穴位注射組(穴注組)、中藥貼敷組(貼敷組)、和穴位注射聯(lián)合中藥貼敷組(穴注+貼敷組),每組12只。復制卵蛋白(OVA)大鼠哮喘模型,分為致敏階段與激發(fā)階段進行。除對照組,其余大鼠分別于第1天、第8天腹腔注射OVA、氫氧化鋁混懸溶液(10%的OVA及10%的氫氧化鋁)1ml致敏,對照組注射同量的生理鹽水。第15天,各組均給予密閉霧化,大鼠置于密閉霧化箱中,以2%OVA溶液超聲霧化激發(fā),對照組以生理鹽水代替,每天一次,每次30min;相比對照組觀察霧化時各組大鼠行為變化,出現(xiàn)煩躁、搔鼻、咳嗽、呼吸頻率加快且幅度加大、明顯的腹式呼吸、點頭呼吸,甚至四肢癱軟俯伏不動、大小便失禁等現(xiàn)象視為激發(fā)成功。連續(xù)激發(fā)7天,后改為每周激發(fā)2次。連續(xù)激發(fā)哮喘發(fā)作7天后,對照組正常喂養(yǎng),其余各組進行治療。取穴大椎、雙定喘、雙肺俞、雙脾俞、雙腎俞,穴注組行草分枝桿菌穴位注射,五組穴位每日輪流取穴注射;貼敷組行貼敷2號穴位貼敷,每貼6h;穴注+貼敷組穴位注射后隨即進行中藥貼敷6h;對照組取穴同治療組,以同量的生理鹽水及空貼代替。隔天操作一次,3次為一療程,共進行四個療程。肺組織HE染色觀察氣道炎癥情況,ELISA法檢測血清Ig E、肺組織勻漿IL-10及IL-17的含量;免疫組織化學(SABC)法檢測肺組織ICAM-1的表達。結果:1各組大鼠行為學表現(xiàn):對照組大鼠呼吸平穩(wěn),霧化時可見蜷縮抱團休息現(xiàn)象,無喘促等特殊表現(xiàn),霧化后精神、飲食正常;模型組大鼠霧化激發(fā)時不同程度出現(xiàn)呼吸頻率快,張口呼吸、點頭呼吸,噴嚏,撓鼻,煩躁不安,腹肌抽動,口鼻及爪甲處紫紺,甚至行動緩慢,癱軟,大小便失禁等,隨激發(fā)次數(shù)增加而癥狀逐步加重,其中由于哮喘發(fā)作死亡1只,數(shù)次激發(fā)后大鼠活動、飲食明顯減少,且有毛色無華,豎毛等表現(xiàn),大鼠模型具備哮喘特征。穴注組、貼敷組、穴注+貼敷組上述癥狀表現(xiàn)不明顯,可見煩躁不安、點頭樣呼吸、搔鼻,但口唇爪甲紫紺較輕,個別可見伏地不動,大小便失禁,但隨激發(fā)次數(shù)減少及治療次數(shù)增多,激發(fā)表現(xiàn)逐漸好轉,穴注+貼敷組好轉較明顯,治療組霧化后飲食活動未見明顯變化。2 HE染色:對照組:細支氣管上皮完整,形態(tài)規(guī)則,支氣管管壁未見增生,肺泡形態(tài)規(guī)則,肺泡壁規(guī)整,未見增生及炎性浸潤。模型組:氣管周圍及肺組織大量炎性細胞浸潤,支氣管狹窄變形,腔內可見脫落上皮細胞,黏膜皺襞增多,黏膜水腫,肺泡壁及肺間隔明顯增厚、充血,肺泡腔明顯變小甚至消失。穴注組:與模型組相比,炎性細胞浸潤減輕,支氣管形態(tài)稍有狹窄等改變,肺泡壁及肺間隔增厚減輕,肺泡形態(tài)欠規(guī)則。貼敷組:與模型組相比,細支氣管形態(tài)較規(guī)則,腔內未見明顯上皮細胞脫落,管壁有炎性細胞浸潤,平滑肌有所增厚,肺泡壁及肺間隔可見增厚減輕,肺泡形態(tài)欠規(guī)則。穴注+貼敷組:細支氣管形態(tài)較規(guī)則,管周可見炎性細胞,管壁輕度增生,黏膜較正常,肺泡腔形態(tài)尚可,個別肺泡壁有所增厚及炎性細胞增多。3 Ig E含量測定結果:模型組血清Ig E明顯高于對照組,有顯著性差異(P0.01);穴注組、貼敷組、穴注+貼敷組三組大鼠的Ig E含量均低于模型組,有顯著差異(P0.01);穴注+貼敷組Ig E含量較穴注組、貼敷組低(P0.05);穴注組與貼敷組Ig E含量無明顯差異(P0.05)。4 IL-10含量測定結果:模型組肺組織IL-10含量明顯低于對照組,有顯著性差異(P0.01),穴注組、貼敷組、穴注+貼敷組三組大鼠的IL-10含量均高于模型組,差異顯著(P0.01),穴注+貼敷組IL-10含量較穴注組、貼敷組高,差異有統(tǒng)計學意義(P0.05),穴注組與貼敷組IL-10含量無明顯差異(P0.05)。5 IL-17含量測定結果:模型組肺組織IL-17含量明顯高于對照組,有顯著性差異(P0.01);與模型組相比,穴注組、貼敷組、穴注+貼敷組三組大鼠的IL-17含量減低,有顯著差異(P0.01);穴注+貼敷組IL-17含量較穴注組、貼敷組低,差異有統(tǒng)計學意義(P0.05),穴注組與貼敷組IL-17含量無明顯差異(P0.05)。6 ICAM-1表達:免疫組化可見,對照組支氣管周圍可見少量陽性細胞;模型組支氣管周圍以及支氣管分泌物及肺間質、肺泡壁棕黃色顆粒表達較多;穴注組和貼敷組支氣管周圍、肺間質肺泡壁棕黃色顆粒較模型組少;穴注+貼敷組支氣管周圍、肺間質、肺泡壁棕黃色陽性細胞表達不明顯。各組平均光密度值比較:模型組肺組織ICAM-1表達明顯高于對照組,有顯著性差異(P0.01),穴注組、貼敷組、穴注+貼敷組三組大鼠的ICAM-1表達均低于模型組,差異顯著(P0.01),穴注+貼敷組ICAM-1表達較穴注組、貼敷組低,差異有統(tǒng)計學意義(P0.05),穴注組與貼敷組ICAM-1表達無明顯差異(P0.05)。結論:1根據模型組大鼠癥狀表現(xiàn)以及各項檢測指標,HE染色提示,支氣管哮喘大鼠造模成功。2支氣管哮喘大鼠血清Ig E的含量增多,肺組織IL-10含量減少、IL-17含量增多,ICAM-1在肺組織的表達增多,并與大鼠激發(fā)哮喘癥狀以及肺臟病理表現(xiàn)變化一致,提示IL-10、IL-17、Ig E、ICAM-1均參與了哮喘的發(fā)病。3穴位注射草分枝桿菌、中藥貼敷可能是通過降低哮喘大鼠Ig E水平,提高哮喘大鼠肺組織中IL-10表達,降低IL-17表達,降低ICAM-1的表達,來減輕氣道炎癥的發(fā)生,調節(jié)免疫平衡,兩種方法綜合應用較單獨應用效果更佳。
[Abstract]:Objective: To observe the effect of acupoint injection of Mycobacterium Mycobacterium and traditional Chinese medicine acupoint application on serum immunoglobulin E (Ig E) level, interleukin 10 (IL-10), interleukin 17 (IL-17), and cell adhesion factor 1 (ICAM-1) expression in the lung tissue, and explore the mechanism of acupuncture point injection and acupoint application of traditional Chinese medicine on bronchial asthma. The clinical application of external treatment for bronchial asthma provides a strong basis and reference. Methods: 60 Wistar male rats of grade SPF were fed for one week. They were randomly divided into control group, model group, acupoint injection group (point injection group), Chinese medicine sticking group (Acupoint Application Group), and Acupoint Injection group (acupoint + adherent group), 12 rats in each group. The asthma model of replicating ovalbumin (OVA) rats was divided into the sensitization stage and the excitation stage. In addition to the control group, the other rats were injected OVA in first days, eighth days in the abdominal cavity, and the aluminum hydroxide suspension solution (10% OVA and 10% aluminum hydroxide) were sensitized by 1ml, and the control group was injected with the same amount of raw saline. All groups were treated with closed atomization for fifteenth days. Rats were placed in close density. In the closed nebulization box, the control group was stimulated by ultrasonic atomization of 2%OVA solution, and the control group was replaced by physiological saline, once a day, each time, 30min each time. Compared with the control group, the behavior changes of rats in each group were observed, such as irritability, scratching nose and coughing, the respiratory rate was accelerated and the amplitude was increased, the obvious abdominal breathing, nodding breathing, even the limbs were paralyzed and inactive, and the size and stool were lost. The forbidden phenomenon was regarded as the excitation success. 7 days of continuous stimulation, then 2 times a week. After 7 days of continuous stimulation of the asthma attack, the control group was fed with normal feeding, the rest of the other groups were treated. Taking the acupoint DZX, double asthma, double lung Yu, double Shenshu, double Shenshu, acupoint injection group acupoint injection of Mycobacterium tumefaciens, the five acupoints were taken every day to take the acupoint injection; the sticking group was put on the patch. 2 acupoint application was applied, each 6h was attached, and 6h was applied after acupoint injection in the acupoint + application group. The control group was taken with the same treatment group, with the same amount of physiological saline and empty paste. 3 times for a course of treatment and four courses of treatment. HE staining of lung tissue was used to observe the airway inflammation, the ELISA method was used to detect the serum Ig E and IL-1 in the lung tissue IL-1. 0 and IL-17 content; immunohistochemistry (SABC) method to detect the expression of ICAM-1 in the lung tissue. Results: 1 the behavior of rats in each group: the control group of rats breathing is stable, the nebulization of the crouching band rest phenomenon, no asthma and other special manifestations, after atomization, the diet is normal; model group rats at different levels of atomization and stimulation of respiratory frequency faster, Breathing, nod, sneezing, nose, irritability, irritability, abdominal muscles twitch, cyanosis in the mouth and the claws, even slow movement, limp, incontinence and so on, the symptoms gradually aggravated with the increase of the number of excuses, 1 of them died of asthma attack, after several times of stimulation, the diet decreased obviously, and the hair color was not Hua, erect hair and so on, and the hair color was not Hua, erect hair and so on. The rat model had the characteristics of asthma. The symptoms of the acupoint injection group, the sticking group and the acupoint injection + cladding group were not obvious, and the symptoms of irritability, nod like breathing, scratching nose were seen, but the cyanosis of the lip and claw was lighter, and the incontinence and incontinence could be seen individually, but the frequency of stimulation and the number of treatment increased, the performance of the stimulation gradually improved, and the acupoint injection + cladding group was better than that. Obviously, there was no obvious change of.2 HE staining in the diet after atomization in the treatment group: the control group: the bronchial epithelium was intact, the shape rule, the bronchial tube wall no hyperplasia, the rules of alveolus shape, the alveolar wall regularity, no hyperplasia and inflammatory infiltration. Exfoliated epithelial cells, mucous plica increased, mucous membrane edema, alveolar wall and pulmonary septum thickening obviously, congestion, alveolar cavity obviously smaller or even disappeared. Group: compared with the model group, inflammatory cells infiltration, bronchi shape slightly narrowing and other changes, alveolar wall and pulmonary septum reduction, alveolar morphology under rules. Patch group: with model group phase The morphology of bronchioles was more regular. There was no obvious epithelial cells falling off in the cavity. The wall of the tube was inflammatory cell infiltration, the smooth muscle was thickened, the alveolar wall and the pulmonary septum were thickening and lessened. The alveolar morphology was under rules. The shape of the bronchioles was more regular. The bronchioles were more regular, the tube wall was slightly proliferated, the mucosa was more normal, and the alveolar cavity was more normal. .3 Ig E content of the serum Ig E in the model group was significantly higher than that of the control group (P0.01). The content of Ig E in three groups of rats in the acupoint injection group, the sticking group and the acupoint injection + cladding group were lower than those in the model group (P0.01), and the Ig E content in the acupoint injection + cladding group was compared with the acupoint injection group. Group low (P0.05); there was no significant difference in the content of Ig E in the acupoint group and the application group (P0.05).4 IL-10 content determination results: the IL-10 content in the lung tissue of the model group was significantly lower than that of the control group (P0.01). The IL-10 content of the three groups of rats in the acupoint injection group, the sticking group and the acupoint injection + cladding group were all higher than the model group, the difference was significant (P0.01), and the IL-10 content of the acupoint injection + application group was significantly different. Compared with the acupoint injection group, the difference was statistically significant (P0.05). There was no significant difference in the content of IL-10 in the acupoint group and the application group (P0.05).5 IL-17 content determination results: the IL-17 content in the lung tissue of the model group was significantly higher than that of the control group (P0.01); the IL-17 content in three groups of rats was compared with the model group, the acupoint injection group, the acupoint injection group and the acupoint injection group. There was a significant difference (P0.01); the content of IL-17 in the acupoint + cladding group was lower than that in the acupoint group, and the difference was statistically significant (P0.05). There was no significant difference (P0.05).6 ICAM-1 expression in the acupoint group and the application group (P0.05).6 ICAM-1 expression: the immunization group was visible, the control group showed a small number of positive cells around the bronchus, and the peripheral bronchus and the bronchial secretion in the model group. The expression of brown and yellow granules in the alveolar wall of the material and pulmonary interstitium was more than that of the model group. The expression of brown yellow granules in the pulmonary interstitial lung wall was less than that in the model group, and the expression of the brown yellow positive cells of the pulmonary interstitial and alveolar wall was not obvious. The average density of the lung tissue in the model group was significantly higher than that in the model group. The ICAM-1 expression in the model group was significantly higher. In the control group, there were significant differences (P0.01). The expression of ICAM-1 in the three groups of rats in the acupoint injection group, the acupoint injection + cladding group were lower than the model group, the difference was significant (P0.01), the ICAM-1 expression in the acupoint injection + cladding group was lower than the acupoint group, and the difference was statistically significant (P0.05). There was no significant difference in the expression of ICAM-1 between the acupoint injection group and the application group (P0.05). Conclusion: 1 basis (P0.05). HE staining showed that the content of Ig E in serum of.2 bronchial asthma rats increased, IL-10 content in lung tissue decreased, IL-17 content increased, and the expression of ICAM-1 increased in lung tissue, and was consistent with the symptoms of irritable asthma and lung pathological changes in rats. IL-10, IL-17, Ig E, ICAM-1 all participated in the.3 point injection of Mycobacterium phlebius in the pathogenesis of asthma. The application of traditional Chinese medicine may be to reduce the expression of IL-10 in the lung tissue of asthmatic rats, reduce the expression of IL-17, reduce ICAM-1 expression, reduce the incidence of airway inflammation and regulate the immune balance. Two methods are used synthetically. The effect of individual application is better.

【學位授予單位】：河北醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R245

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